Jirapat Namkaew, Jun Zhang, Norio Yamakawa, Yoshimasa Hamada, Kazue Tsugawa, Miho Oyadomari, Masato Miyake, Toyomasa Katagiri and Seiichi Oyadomari : Repositioning of mifepristone as an integrated stress response activator to potentiate cisplatin efficacy in non-small cell lung cancer., Cancer Letters, Vol.582, 2023.
(要約)
Lung cancer, primarily non-small-cell lung cancer (NSCLC), is a significant cause of cancer-related mortality worldwide. Cisplatin-based chemotherapy is a standard treatment for NSCLC; however, its effectiveness is often limited due to the development of resistance, leading to NSCLC recurrence. Thus, the identification of effective chemosensitizers for cisplatin is of paramount importance. The integrated stress response (ISR), activated by various cellular stresses and mediated by eIF2α kinases, has been implicated in drug sensitivity. ISR activation globally suppresses protein synthesis while selectively promoting the translation of ATF4 mRNA, which can induce pro-apoptotic proteins such as CHOP, ATF3, and TRIB3. To expedite and economize the development of chemosensitizers for cisplatin treatment in NSCLC, we employed a strategy to screen an FDA-approved drug library for ISR activators. In this study, we identified mifepristone as a potent ISR activator. Mifepristone activated the HRI/eIF2α/ATF4 axis, leading to the induction of pro-apoptotic factors, independent of its known role as a synthetic steroid. Our in vitro and in vivo models demonstrated mifepristone's potential to inhibit NSCLC re-proliferation following cisplatin treatment and tumor growth, respectively, via the ISR-mediated cell death pathway. These findings suggest that mifepristone, as an ISR activator, could enhance the efficacy of cisplatin-based therapy for NSCLC, highlighting the potential of drug repositioning in the search for effective chemosensitizers.
(キーワード)
Humans / Carcinoma, Non-Small-Cell Lung / Cisplatin / Lung Neoplasms / Mifepristone / Drug Repositioning / Signal Transduction / Cell Line, Tumor / Drug Resistance, Neoplasm
Atsushi Saito, Yasunao Kamikawa, Taichi Ito, Koji Matsuhisa, Masayuki Kaneko, Takumi Okamoto, Tetsuro Yoshimaru, Yosuke Matsushita, Toyomasa Katagiri and Kazunori Imaizumi : p53-independent tumor suppression by cell-cycle arrest via CREB/ATF transcription factor OASIS, Cell Reports, Vol.42, No.5, 112479, 2023.
(要約)
CREB/ATF transcription factor OASIS/CREB3L1 is upregulated in long-term-cultured astrocytes undergoing cell-cycle arrest due to loss of DNA integrity by repeated replication. However, the roles of OASIS in the cell cycle remain unexplored. We find that OASIS arrests the cell cycle at G/M phase after DNA damage via direct induction of p21. Cell-cycle arrest by OASIS is dominant in astrocytes and osteoblasts, but not in fibroblasts, which are dependent on p53. In a brain injury model, Oasis reactive astrocytes surrounding the lesion core show sustained growth and inhibition of cell-cycle arrest, resulting in prolonged gliosis. We find that some glioma patients exhibit low expression of OASIS due to high methylation of its promoter. Specific removal of this hypermethylation in glioblastomas transplanted into nude mice by epigenomic engineering suppresses the tumorigenesis. These findings suggest OASIS as a critical cell-cycle inhibitor with potential to act as a tumor suppressor.
Kosaku Okuda, Kengo Nakahara, Akihiro Ito, Yuta Iijima, Ryosuke Nomura, Ashutosh Kumar, Kana Fujikawa, Kazuya Adachi, Yuki Shimada, Satoshi Fujio, Reina Yamamoto, Nobumasa Takasugi, Kunishige Onuma, Mitsuhiko Osaki, Futoshi Okada, Taichi Ukegawa, Yasuo Takeuchi, Norihisa Yasui, Atsuko Yamashita, Hiroyuki Marusawa, Yosuke Matsushita, Toyomasa Katagiri, Takahiro Shibata, Koji Uchida, Sheng-Yong Niu, B Nhi Lang, Tomohiro Nakamura, J Kam Y Zhang, A Stuart Lipton and Takashi Uehara : Pivotal role for S-nitrosylation of DNA methyltransferase 3B in epigenetic regulation of tumorigenesis., Nature Communications, Vol.14, No.1, 2023.
(要約)
≤ 100 nM). Treatment with DBIC prevents nitric oxide (NO)-induced conversion of human colonic adenoma to adenocarcinoma in vitro. Additionally, in vivo treatment with DBIC strongly attenuates tumor development in a mouse model of carcinogenesis triggered by inflammation-induced generation of NO. Our results demonstrate that de novo DNA methylation mediated by DNMT3B is regulated by NO, and DBIC protects against tumor formation by preventing aberrant S-nitrosylation of DNMT3B.
(キーワード)
Animals / Humans / Mice / Cell Transformation, Neoplastic / DNA (Cytosine-5-)-Methyltransferase 1 / DNA (Cytosine-5-)-Methyltransferases / DNA Methylation / DNA Modification Methylases / Epigenesis, Genetic
Makoto Takeuchi, Toshihiko Nishisho, Shun-ichi Toki, Shinji Kawaguchi, Shunsuke Tamaki, Takeshi Oya, Yoshihiro Uto, Toyomasa Katagiri and Koichi Sairyo : Blue light induces apoptosis and autophagy by promoting ROS-mediated mitochondrial dysfunction in synovial sarcoma., Cancer Medicine, Vol.12, No.8, 9668-9683, 2023.
(要約)
Taken together, our results revealed that BL induced apoptosis via the ROS-mitochondrial signaling pathway, and autophagy was activated in response to the production of ROS, which protected SS cells from apoptosis. Therefore, BL is a promising candidate for the development of an antitumor therapeutic strategy targeting SS.
Sayumi Fujimori, Izumi Ohigashi, Hayato Abe, Yosuke Matsushita, Toyomasa Katagiri, Taketo M. Makoto, Takahama Yousuke and Takada Shinji : Fine-tuning of β-catenin in mouse thymic epithelial cells is required for postnatal T-cell development, eLife, Vol.11, e69088, 2022.
(要約)
In the thymus, the thymic epithelium provides a microenvironment essential for the development of functionally competent and self-tolerant T cells. Previous findings showed that modulation of Wnt/β-catenin signaling in mouse thymic epithelial cells (TECs) disrupts embryonic thymus organogenesis. However, the role of β-catenin in TECs for postnatal T-cell development remains to be elucidated. Here, we analyzed gain-of-function (GOF) and loss-of-function (LOF) of β-catenin highly specific in mouse TECs. We found that GOF of β-catenin in TECs results in severe thymic dysplasia and T-cell deficiency beginning from the embryonic period. By contrast, LOF of β-catenin in TECs reduces the number of cortical TECs and thymocytes modestly and only postnatally. These results indicate that fine-tuning of β-catenin expression within a permissive range is required for TECs to generate an optimal microenvironment to support postnatal T-cell development.
Hitomi Sudo, B Atsushi Tsuji, Aya Sugyo, Yosuke Harada, Satoshi Nagayama, Toyomasa Katagiri, Yusuke Nakamura and Tatsuya Higashi : FZD10-targeted α-radioimmunotherapy with 225 Ac-labeled OTSA101 achieves complete remission in a synovial sarcoma model, Cancer Science, Vol.113, No.2, 721-732, 2021.
(要約)
Synovial sarcomas are rare tumors arising in adolescents and young adults. The prognosis for advanced disease is poor, with an overall survival of 12-18 months. Frizzled homolog 10 (FZD10) is overexpressed in most synovial sarcomas, making it a promising therapeutic target. The results of a phase 1 trial of β-radioimmunotherapy (RIT) with the
Shun-ichi Toki, Tetsuro Yoshimaru, Yosuke Matsushita, Hitoshi Aibara, Masaya Ono, Koichi Tsuneyama, Koichi Sairyo and Toyomasa Katagiri : The survival and proliferation of osteosarcoma cells are dependent on the mitochondrial BIG3-PHB2 complex formation., Cancer Science, Vol.112, No.10, 4208-4219, 2021.
(要約)
Previous studies reported the critical role of the brefeldin A-inhibited guanine nucleotide exchange protein 3-prohibitin 2 (BIG3-PHB2) complex in modulating estrogen signaling activation in breast cancer cells, yet its pathophysiological roles in osteosarcoma (OS) cells remain elusive. Here, we report a novel function of BIG3-PHB2 in OS malignancy. BIG3-PHB2 complexes were localized mainly in mitochondria in OS cells, unlike in estrogen-dependent breast cancer cells. Depletion of endogenous BIG3 expression by small interfering RNA (siRNA) treatment led to significant inhibition of OS cell growth. Disruption of BIG3-PHB2 complex formation by treatment with specific peptide inhibitor also resulted in significant dose-dependent suppression of OS cell growth, migration, and invasion resulting from G2/M-phase arrest and in PARP cleavage, ultimately leading to PARP-1/apoptosis-inducing factor (AIF) pathway activation-dependent apoptosis in OS cells. Subsequent proteomic and bioinformatic pathway analyses revealed that disruption of the BIG3-PHB2 complex might lead to downregulation of inner mitochondrial membrane protein complex activity. Our findings indicate that the mitochondrial BIG3-PHB2 complex might regulate PARP-1/AIF pathway-dependent apoptosis during OS cell proliferation and progression and that disruption of this complex may be a promising therapeutic strategy for OS.
Tetsuro Yoshimaru, Yusuke Nakamura and Toyomasa Katagiri : Functional genomics for breast cancer drug target discovery., Journal of Human Genetics, Vol.66, No.9, 927-935, 2021.
(要約)
Breast cancer is a heterogeneous disease that develops through a multistep process via the accumulation of genetic/epigenetic alterations in various cancer-related genes. Current treatment options for breast cancer patients include surgery, radiotherapy, and chemotherapy including conventional cytotoxic and molecular-targeted anticancer drugs for each intrinsic subtype, such as endocrine therapy and antihuman epidermal growth factor receptor 2 (HER2) therapy. However, these therapies often fail to prevent recurrence and metastasis due to resistance. Overall, understanding the molecular mechanisms of breast carcinogenesis and progression will help to establish therapeutic modalities to improve treatment. The recent development of comprehensive omics technologies has led to the discovery of driver genes, including oncogenes and tumor-suppressor genes, contributing to the development of molecular-targeted anticancer drugs. Here, we review the development of anticancer drugs targeting cancer-specific functional therapeutic targets, namely, MELK (maternal embryonic leucine zipper kinase), TOPK (T-lymphokine-activated killer cell-originated protein kinase), and BIG3 (brefeldin A-inhibited guanine nucleotide-exchange protein 3), as identified through comprehensive breast cancer transcriptomics.
(キーワード)
Antineoplastic Agents / Breast Neoplasms / Drug Discovery / Female / Genomics / Humans
Takashi Sugiyama, Naoya Murao, Hisae Kadowaki, Keizo Takao, Tsuyoshi Miyakawa, Yosuke Matsushita, Toyomasa Katagiri, Akira Futatsugi, Yohei Shinmyo, Hiroshi Kawasaki, Juro Sakai, Kazutaka Shiomi, Masamitsu Nakazato, Kohsuke Takeda, Katsuhiko Mikoshiba, L Hidde Ploegh, Hidenori Ichijo and Hideki Nishitoh : ERAD components Derlin-1 and Derlin-2 are essential for postnatal brain development and motor function., iScience, Vol.24, No.7, 2021.
(要約)
and surprisingly also inhibited sterol regulatory element binding protein 2 (SREBP-2)-mediated brain cholesterol biosynthesis. In addition, reduced neurite outgrowth due to Derlin-1 deficiency was rescued by SREBP-2 pathway activation. Overall, our findings demonstrate that Derlins sustain brain cholesterol biosynthesis, which is essential for appropriate postnatal brain development and function.
). East Asian-specific missense variants were identified as candidate causal variants for three novel loci, and we successfully replicated two of them by analyzing independent Japanese cohorts; p.R220W of ATG16L2 (associated with coronary artery disease) and p.V326A of POT1 (associated with lung cancer). We further investigated enrichment of heritability within 2,868 annotations of genome-wide transcription factor occupancy, and identified 378 significant enrichments across nine diseases (false discovery rate < 0.05) (for example, NKX3-1 for prostate cancer). This large-scale GWAS in a Japanese population provides insights into the etiology of complex diseases and highlights the importance of performing GWAS in non-European populations.
(キーワード)
Cohort Studies / Female / Genetic Predisposition to Disease / Genetic Variation / Genome-Wide Association Study / Humans / Inheritance Patterns / Japan / Male / Sex Factors / Transcription Factors
Yukie Fujimoto, Natsuko Inoue, Koji Morimoto, Takahiro Watanabe, Seiichi Hirota, Michiko Imamura, Yosuke Matsushita, Toyomasa Katagiri, Haruki Okamura and Yasuo Miyoshi : Significant association between high serum CCL5 levels and better disease-free survival of patients with early breast cancer., Cancer Science, Vol.111, No.1, 209-218, 2019.
(要約)
Analysis of anticancer immunity aids in assessing the prognosis of patients with breast cancer. From 250 operated breast cancers, we focused on serum levels of C-C motif chemokine ligand 5 (CCL5), which is involved in cancer immune reactions. Serum levels of CCL5 were measured using a cytometric bead-based immunoassay kit and CCL5 expression in cancer cells was determined using immunohistochemical staining. In addition, mRNA in cancer and stromal cells was analyzed by microdissection and comparison with the public dataset. Disease-free survival (DFS) of patients with high CCL5 levels (cut-off, 13.87 ng/mL; n = 192) was significantly better than those with low CCL5 levels (n = 58; hazard ratio, 0.20; 95% confidence interval, 0.10-0.39; P < .0001). An improved overall survival was observed in patients with high CCL5 levels compared to those with low CCL5 levels (P = .024). On the contrary, high immunohistochemical expression of CCL5 in cancer cells was significantly associated with decreased DFS. As serum CCL5 levels did not correlate with CCL5 expression in cancer cells and the relative expression of mRNA CCL5 was elevated in stromal cells in relation to cancer cells, serum CCL5 might be derived not from cancer cells, but from stromal cells. Expression of CCL5 in serum, but not in cancer cells, might contribute to improved patient prognosis mediating through not only immune reaction, but through other mechanisms. Determination of circulating CCL5 levels could be useful for predicting patient prognosis.
Ryuichiro Kimura, Tetsuro Yoshimaru, Yosuke Matsushita, Taisuke Matsuo, Masaya Ono, Jae-Hyun Park, Mitsunori Sasa, Yasuo Miyoshi, Yusuke Nakamura and Toyomasa Katagiri : The GALNT6‑LGALS3BP axis promotes breast cancer cell growth., International Journal of Oncology, Vol.56, No.2, 581-595, 2019.
(要約)
Polypeptide N‑acetylgalactosaminyltransferase 6 (GALNT6), which is involved in the initiation of O‑glycosylation, has been reported to play crucial roles in mammary carcinogenesis through binding to several substrates; however, its biological roles in mediating growth‑promoting effects remain unknown. The present study demonstrated a crucial pathophysiological role of GALNT6 through its O‑glycosylation of lectin galactoside‑binding soluble 3 binding protein (LGALS3BP), a secreted growth‑promoting glycoprotein, in breast cancer growth. The Cancer Genome Atlas data analysis revealed that high expression levels of GALNT6 were significantly associated with poor prognosis of breast cancer. GALNT6 O‑glycosylated LGALS3BP in breast cancer cells, whereas knockdown of GALNT6 by siRNA led to the inhibition of both the O‑glycosylation and secretion of LGALS3BP, resulting in the suppression of breast cancer cell growth. Notably, LGALS3BP is potentially O‑glycosylated at three sites (T556, T571 and S582) by GALNT6, thereby promoting autocrine cell growth, whereas the expression of LGALS3BP with three Ala substitutions (T556A, T571A and S582A) in cells drastically reduced GALNT6‑dependent LGALS3BP O‑glycosylation and secretion, resulting in suppression of autocrine growth‑promoting effect. The findings of the present study suggest that the GALNT6‑LGALS3BP axis is crucial for breast cancer cell proliferation and may be a therapeutic target and biomarker for mammary tumors.
Siew-Kee Low, Ming Yoon Chin, Hidemi Ito, Keitaro Matsuo, Chizu Tanikawa, Koichi Matsuda, Hiroko Saito, Mika Sakurai-Yageta, Naoki Nakaya, Atsushi Shimizu, S Satoshi Nishizuka, Taiki Yamaji, Norie Sawada, Motoki Iwasaki, Shoichiro Tsugane, Toshiro Takezaki, Sadao Suzuki, Mariko Naito, Kenji Wakai, Yoichiro Kamatani, Yukihide Momozawa, Yoshinori Murakami, Johji Inazawa, Yusuke Nakamura, Michiaki Kubo, Toyomasa Katagiri and Yoshio Miki : Identification of two novel breast cancer loci through large-scale genome-wide association study in the Japanese population., Scientific Reports, Vol.9, No.1, 17332, 2019.
(要約)
Genome-wide association studies (GWAS) have successfully identified about 70 genomic loci associated with breast cancer. Owing to the complexity of linkage disequilibrium and environmental exposures in different populations, it is essential to perform regional GWAS for better risk prediction. This study aimed to investigate the genetic architecture and to assess common genetic risk model of breast cancer with 6,669 breast cancer patients and 21,930 female controls in the Japanese population. This GWAS identified 11 genomic loci that surpass genome-wide significance threshold of P < 5.0 × 10
Natsuko Inoue, Wen Li, Yukie Fujimoto, Yosuke Matsushita, Toyomasa Katagiri, Haruki Okamura and Yasuo Miyoshi : High Serum Levels of Interleukin-18 Are Associated With Worse Outcomes in Patients With Breast Cancer., Anticancer Research, Vol.39, No.9, 5009-5018, 2019.
(要約)
IL-18 levels may be useful for predicting the prognosis of patients who have received surgical treatment for breast cancer.
Takeru Chigira, Satoru Nagatoishi, Hiroyuki Takeda, Tetsuro Yoshimaru, Toyomasa Katagiri and Kouhei Tsumoto : Biophysical characterization of the breast cancer-related BIG3-PHB2 interaction: Effect of non-conserved loop region of BIG3 on the structure and the interaction., Biochemical and Biophysical Research Communications, Vol.518, No.1, 183-189, 2019.
(要約)
Brefeldin A-inhibited guanine nucleotide-exchange protein 3 (BIG3) interacts with and inhibits the tumor suppressor function of prohibitin-2 (PHB2), and recent in vivo studies have demonstrated that the BIG3-PHB2 interaction is a promising target for breast cancer therapy. However, little biophysical characterization on BIG3 and its interaction with PHB2 has been reported. Here we compared the calculated 8-class secondary structure of the N-terminal domains of BIG family proteins and identified a loop region unique to BIG3. Our biophysical characterization demonstrated that this loop region significantly affects the colloidal and thermodynamic stability of BIG3 and the thermodynamic and kinetic profile of its interaction with PHB2. These results establish a model for the BIG3-PHB2 interaction and an entry for drug discovery for breast cancer.
Yukihide Momozawa, Yusuke Iwasaki, T Michael Parsons, Yoichiro Kamatani, Atsushi Takahashi, Chieko Tamura, Toyomasa Katagiri, Teruhiko Yoshida, Seigo Nakamura, Kokichi Sugano, Yoshio Miki, Makoto Hirata, Koichi Matsuda, B Amanda Spurdle and Michiaki Kubo : Germline pathogenic variants of 11 breast cancer genes in 7,051 Japanese patients and 11,241 controls., Nature Communications, Vol.9, No.1, 2018.
(要約)
Pathogenic variants in highly penetrant genes are useful for the diagnosis, therapy, and surveillance for hereditary breast cancer. Large-scale studies are needed to inform future testing and variant classification processes in Japanese. We performed a case-control association study for variants in coding regions of 11 hereditary breast cancer genes in 7051 unselected breast cancer patients and 11,241 female controls of Japanese ancestry. Here, we identify 244 germline pathogenic variants. Pathogenic variants are found in 5.7% of patients, ranging from 15% in women diagnosed <40 years to 3.2% in patients 80 years, with BRCA1/2, explaining two-thirds of pathogenic variants identified at all ages. BRCA1/2, PALB2, and TP53 are significant causative genes. Patients with pathogenic variants in BRCA1/2 or PTEN have significantly younger age at diagnosis. In conclusion, BRCA1/2, PALB2, and TP53 are the major hereditary breast cancer genes, irrespective of age at diagnosis, in Japanese women.
Boya Deng, Emre Yunus Tarhan, Koji Ueda, Lili Ren, Toyomasa Katagiri, Jae-Hyun Park and Yusuke Nakamura : Critical Role of Estrogen Receptor Alpha O-Glycosylation by N-Acetylgalactosaminyltransferase 6 (GALNT6) in Its Nuclear Localization in Breast Cancer Cells., Neoplasia, Vol.20, No.10, 1038-1044, 2018.
(要約)
Alteration of protein O-glycosylation in various human cancers including breast cancer is well known, but molecular roles of their aberrant glycosylations on cancer have not been fully understood. We previously reported critical roles of polypeptide N-acetylgalactosaminyltransferase 6 (GALNT6 or GalNAc-T6) that was upregulated in a great majority of breast cancer tissues. Here we further report O-glycosylation of estrogen receptor alpha (ER-) by GALNT6 and the significant role of its nuclear localization in breast cancer cells. Knockdown of GALNT6 expression in two breast cancer cell lines, T47D and MCF7, in which both ER- and GALNT6 were highly expressed, by small interfering RNA could significantly attenuate expression of ER-. Immunocytochemical analysis clearly demonstrated the drastic decrease of ER- protein in the nucleus of these cancer cells. Accordingly, the downstream genes of the ER- pathway such as MYC, CCND1, and CTSD were significantly downregulated. We confirmed GALNT6-dependent ER- O-glycosylation and identified O-glycosylation of S573 in an F domain of ER- by GALNT6 through LC-MS/MS analysis. We also obtained evidences showing that the glycosylation of ER- at S573 by GALNT6 is essential for protein stability and nuclear localization of ER- in breast cancer cells. Furthermore, we designed cell membrane-permeable peptides including the O-glycosylation site and found a significant decrease of the cell viability of breast cancer cells by treatment of these peptides in a GALNT6 expression-dependent manner. Our study suggests that targeting the GALNT6 enzymatic activity as well as the GALNT6/ER- interaction could be a promising therapeutic approach to ER--positive breast cancer patients.
Yoichiro Kato, Hitoshi Zembutsu, Ryo Takata, Tomohiko Matsuura, Renpei Kato, Mitsugu Kanehira, Kazuhiro Iwasaki, Noriyuki Yamada, Toyomasa Katagiri, Tamotsu Sugai, Tomoaki Fujioka, Yusuke Nakamura and Wataru Obara : A prospective study to examine the accuracies and efficacies of prediction systems for response to neoadjuvant chemotherapy for muscle invasive bladder cancer., Oncology Letters, Vol.16, No.5, 5775-5784, 2018.
(要約)
The present study established systems to predict the chemo-sensitivity of muscle invasive bladder cancer (MIBC) for neoadjuvant chemotherapy (NAC) with methotrexate, vinblastine, doxorubicin plus cisplatin (M-VAC) and carboplatin plus gemcitabine (CaG) by analyzing microarray data. The primary aim of the study was to investigate whether the clinical response would increase by combining these prediction systems. Treatment of each MIBC case was allocated into M-VAC NAC, CaG NAC, surgery, or radiation therapy groups by their prediction score (PS), which was calculated using the designed chemo-sensitivity prediction system. The therapeutic effect of the present study was compared with the results of historical controls (n=76 patients) whose treatments were not allocated using the chemo-sensitivity prediction system. In addition, the overall survival between the predicted to be responder (positive PS) group and predicted to be non-responder (negative PS) group was investigated in the present study. Of the 33 patients with MIBC, 25 cases were positive PS and 8 were negative PS. Among the 25 positive PS cases, 7 were allocated to receive M-VAC NAC and 18 were allocated to receive CaG NAC according to the results of the prediction systems. Of the 8 negative PS cases, 3 received CaG NAC, 1 received surgery without NAC and 4 received radiation therapy. The total clinical response to NAC was 88.0% (22/25), which was significantly increased compared with the historical controls [56.6% (43/76) P=0.0041]. Overall survival of the positive PS group in the study was significantly increased compared with the negative PS group (P=0.027). In conclusion, the combination of the two prediction systems may increase the treatment efficacy for patients with MIBC by proposing the optimal NAC regimen. In addition, the positive PS group would have a better prognosis compared with the negative PS group. These results suggest that the two prediction systems may lead to the achievement of 'precision medicine'.
Anne-Laure Giraudet, Alexandre Philippe Cassier, Chicaco Iwao-Fukukawa, Gwenaelle Garin, Jean-Noël Badel, David Kryza, Sylvie Chabaud, Laurence Gilles-Afchain, Gilles Clapisson, Claude Desuzinges, David Sarrut, Adrien Halty, Antoine Italiano, Masaharu Mori, Takuya Tsunoda, Toyomasa Katagiri, Yusuke Nakamura, Laurent Alberti, Claire Cropet, Simon Baconnier, Sandrine Berge-Montamat, David Pérol and Jean-Yves Blay : A first-in-human study investigating biodistribution, safety and recommended dose of a new radiolabeled MAb targeting FZD10 in metastatic synovial sarcoma patients., BMC Cancer, Vol.18, No.1, 646, 2018.
(要約)
Y-OTSA-101 treatment. The recommended activity for further clinical investigations is 1110 MBq of
Kei Daizumoto, Tetsuro Yoshimaru, Yosuke Matsushita, Tomoya Fukawa, Hisanori Uehara, Masaya Ono, Masato Komatsu, Hiro-omi Kanayama and Toyomasa Katagiri : A DDX31/mutant-p53/EGFR axis promotes multistep progression of muscle invasive bladder cancer, Cancer Research, Vol.78, No.9, 2233-2247, 2018.
(要約)
The p53 and EGFR pathways are frequently altered in bladder cancer, yet their contributions to its progression remain elusive. Here we report that DEAD box polypeptide 31 (DDX31) plays a critical role in the multistep progression of muscle-invasive bladder cancer (MIBC) through its sequential interactions with mutant p53 (mutp53) and EGFR. In early MIBC cells, nuclear DDX31-bound mutp53/SP1 enhanced mutp53 transcriptional activation, leading to migration and invasion of MIBC. Cytoplasmic DDX31 also bound EGFR and phospho-nucleolin in advanced MIBC, leading to EGFR-Akt signaling activation. High expression of both cytoplasmic DDX31 and p53 proteins correlated with poor prognosis in patients with MIBC, and blocking the DDX31/NCL interaction resulted in downregulation of EGFR/Akt signaling, eliciting an antitumor effect against bladder cancer. These findings reveal that DDX31 cooperates with mutp53 and EGFR to promote progression of MIBC, and inhibition of DDX31/NCL formation may lead to potential treatment strategies for advanced MIBC. DDX31 cooperates with mutp53 and EGFR to promote progression of muscle invasive bladder cancer. .
Yoshimasa Miyagawa, Yosuke Matsushita, Hiromu Suzuki, Masato Komatsu, Tetsuro Yoshimaru, Ryuichiro Kimura, Ayako Yanai, Junko Honda, Akira Tangoku, Mitsunori Sasa, Yasuo Miyoshi and Toyomasa Katagiri : Frequent downregulation of LRRC26 by epigenetic alterations is involved in the malignant progression of triple-negative breast cancer., International Journal of Oncology, 2018.
(要約)
Triple-negative breast cancer (TNBC), defined as breast cancer lacking estrogen- and progesterone‑receptor expression and human epidermal growth factor receptor 2 (HER2) amplification, is a heterogeneous disease. RNA-sequencing analysis of 15 TNBC specimens and The Cancer Genome Atlas-TNBC dataset analysis identified the frequent downregulation of leucine-rich repeat-containing 26 (LRRC26), which negatively regulates nuclear factor-κB (NF-κB) signaling, in TNBC tissues. Quantitative polymerase chain reaction and bisulfite pyrosequencing analyses revealed that LRRC26 was frequently silenced in TNBC tissues and cell lines as a result of promoter methylation. LRRC26 expression was restored by 5-aza-2'-deoxycytidine (5'-aza-dC) treatment in HCC1937 TNBC cells, which lack LRRC26 expression. Notably, small interfering RNA-mediated knockdown of LRRC26 expression significantly enhanced the anchorage-independent growth, invasion and migration of HCC70 cells, whereas ectopic overexpression of LRRC26 in BT20 cells suppressed their invasion and migration. Conversely, neither knockdown nor overexpression of LRRC26 had an effect on cell viability in the absence of tumor necrosis factor-α (TNF-α) stimulation. Meanwhile, overexpression of LRRC26 caused the reduction of TNF-α-mediated NF-κB luciferase reporter activity, whereas depleting LRRC26 expression resulted in the upregulation of TNF-α-mediated NF-κB downstream genes [interleukin-6 (IL-6), IL-8 and C-X-C motif chemokine ligand-1]. Taken together, these findings demonstrate that LRRC26 is frequently downregulated in TNBC due to DNA methylation and that it suppresses the TNF-α-independent anchorage-independent growth, invasion and migration of TNBC cells. Loss of LRRC26 function may be a critical event in the aggressiveness of TNBC cells through a TNF-α/NF-κB-independent mechanism.
Wataru Obara, Mitsugu Kanehira, Toyomasa Katagiri, Renpei Kato, Yoichiro Kato and Ryo Takata : Present status and future perspective of peptide-based vaccine therapy for urological cancer., Cancer Science, Vol.109, No.3, 550-559, 2018.
(要約)
Use of peptide-based vaccines as therapeutics aims to elicit immune responses through antigenic epitopes derived from tumor antigens. Peptide-based vaccines are easily synthesized and lack significant side-effects when given in vivo. Peptide-based vaccine therapy against several cancers including urological cancers has made progress for several decades, but there is no worldwide approved peptide vaccine. Peptide vaccines were also shown to induce a high frequency of immune response in patients accompanied by clinical efficacy. These data are discussed in light of the recent progression of immunotherapy caused by the addition of immune checkpoint inhibitors thus providing a general picture of the potential therapeutic efficacy of peptide-based vaccines and their combination with other biological agents. In this review, we discuss the mechanism of the antitumor effect of peptide-based vaccine therapy, development of our peptide vaccine, recent clinical trials using peptide vaccines for urological cancers, and perspectives of peptide-based vaccine therapy.
Arisa Nishimukai, Natsuko Inoue, Ayako Kira, Masashi Takeda, Koji Morimoto, Kazuhiro Araki, Kazuhiro Kitajima, Takahiro Watanabe, Seiichi Hirota, Toyomasa Katagiri, Shoji Nakamori, Kouhei Akazawa and Yasuo Miyoshi : Tumor size and proliferative marker geminin rather than Ki67 expression levels significantly associated with maximum uptake of 18F-deoxyglucose levels on positron emission tomography for breast cancers., PLoS ONE, Vol.12, No.9, 2017.
(要約)
It has been well established that maximum standardized uptake value (SUVmax) for 18F-fluorodeoxyglucose positron-emission tomography/computed tomography (FDG PET/CT) is clinically useful for evaluating treatment efficacy as well as predicting prognosis of breast cancer patients. Although SUVmax reflects increased glucose uptake and metabolism possibly induced by activation of growth factor signaling or TP53 dysfunction, tumor characteristics of SUVmax-high breast cancers remain to be elucidated. For the present study, we used immunohistochemical staining to investigate expressions of phospho-ribosomal protein S6 (pS6, downstream molecule of phosphatidyl inositol 3-kinase/Akt/mammalian target of the rapamycin/S6K pathway) and phosphor-p44/42 mitogen-activated protein kinase (pMAPK). Expression levels of TP53 and proliferative marker geminin as well as Ki67 were also examined by means of immunostaining in 163 invasive breast cancers. Cutoff values were set at 10% for pS6, 20% for pMAPK and TP53, and 4% for geminin. The SUVmax levels were significantly higher in the pS6-positive (p = 0.0173), TP53-positive (p = 0.0207) and geminin-high cancers (p<0.0001), but there was no significant association between pMAPK expression levels and SUVmax levels. Multivariable analysis showed that a high geminin level (odds ratio: 6.497, 95% confidence interval: 2.427-19.202, p = 0.0001) and large tumor size (6.438, 2.224-20.946, p = 0.0005) were significantly and independently associated with SUVmax-high. Univariable but not multivariable analysis indicated that Ki67-high significantly correlated with SUVmax-high. Twenty of 23 (87.0%) breast cancers with tumor size >2cm and geminin-high showed SUVmax-high, while only 6 of 49 (12.2%) breast cancers 2cm in size and with low geminin levels were SUVmax-high. In conclusion, we could determine that breast cancers with a large tumor and a geminin-high rather than Ki67-high proliferative marker were significantly associated with high levels of SUVmax. These findings may signify that SUVmax reflects tumor characteristics with high proliferative activity but not activation of mTOR/S6K and MAPK pathways or increased glucose metabolism due to dysfunction of TP53.
Tetsuro Yoshimaru, Masaya Ono, Yoshimi Bando, Yi-An Chen, Kenji Mizuguchi, Hiroshi Shima, Masato Komatsu, Issei Imoto, Keisuke Izumi, Junko Honda, Yasuo Miyoshi, Mitsunori Sasa and Toyomasa Katagiri : A-kinase anchoring protein BIG3 coordinates oestrogen signalling in breast cancer cells., Nature Communications, Vol.8, No.15427, 2017.
(要約)
Approximately 70% of breast cancer cells express oestrogen receptor alpha (ER). Previous studies have shown that the Brefeldin A-inhibited guanine nucleotide-exchange protein 3-prohibitin 2 (BIG3-PHB2) complex has a crucial role in these cells. However, it remains unclear how BIG3 regulates the suppressive activity of PHB2. Here we demonstrate that BIG3 functions as an A-kinase anchoring protein that binds protein kinase A (PKA) and the isoform of the catalytic subunit of protein phosphatase 1 (PP1C), thereby dephosphorylating and inactivating PHB2. E2-induced PKA-mediated phosphorylation of BIG3-S305 and -S1208 serves to enhance PP1C activity, resulting in E2/ER signalling activation via PHB2 inactivation due to PHB2-S39 dephosphorylation. Furthermore, an analysis of independent cohorts of ER-positive breast cancers patients reveal that both BIG3 overexpression and PHB2-S39 dephosphorylation are strongly associated with poor prognosis. This is the first demonstration of the mechanism of E2/ER signalling activation via the BIG3-PKA-PP1C tri-complex in breast cancer cells.
Daichi Shigemizu, Takuji Iwase, Masataka Yoshimoto, Yasuyo Suzuki, Fuyuki Miya, A Keith Boroevich, Toyomasa Katagiri, Hitoshi Zembutsu and Tatsuhiko Tsunoda : The prediction models for postoperative overall survival and disease-free survival in patients with breast cancer., Cancer Medicine, Vol.6, No.7, 1627-1638, 2017.
(要約)
The goal of this study is to establish a method for predicting overall survival (OS) and disease-free survival (DFS) in breast cancer patients after surgical operation. The gene expression profiles of cancer tissues from the patients, who underwent complete surgical resection of breast cancer and were subsequently monitored for postoperative survival, were analyzed using cDNA microarrays. We detected seven and three probes/genes associated with the postoperative OS and DFS, respectively, from our discovery cohort data. By incorporating these genes associated with the postoperative survival into MammaPrint genes, often used to predict prognosis of patients with early-stage breast cancer, we constructed postoperative OS and DFS prediction models from the discovery cohort data using a Cox proportional hazard model. The predictive ability of the models was evaluated in another independent cohort using Kaplan-Meier (KM) curves and the area under the receiver operating characteristic curve (AUC). The KM curves showed a statistically significant difference between the predicted high- and low-risk groups in both OS (log-rank trend test P = 0.0033) and DFS (log-rank trend test P = 0.00030). The models also achieved high AUC scores of 0.71 in OS and of 0.60 in DFS. Furthermore, our models had improved KM curves when compared to the models using MammaPrint genes (OS: P = 0.0058, DFS: P = 0.00054). Similar results were observed when our model was tested in publicly available datasets. These observations indicate that there is still room for improvement in the current methods of predicting postoperative OS and DFS in breast cancer.
Estradiol (E2) and the oestrogen receptor-alpha (ER) signalling pathway play pivotal roles in the proliferative activity of breast cancer cells. Recent findings show that the brefeldin A-inhibited guanine nucleotide-exchange protein 3-prohibitin 2 (BIG3-PHB2) complex plays a crucial role in E2/ER signalling modulation in breast cancer cells. Moreover, specific inhibition of the BIG3-PHB2 interaction using the ER activity-regulator synthetic peptide (ERAP: 165-177 amino acids), derived from -helical BIG3 sequence, resulted in a significant anti-tumour effect. However, the duration of this effect was very short for viable clinical application. We developed the chemically modified ERAP using stapling methods (stapledERAP) to improve the duration of its antitumour effects. The stapledERAP specifically inhibited the BIG3-PHB2 interaction and exhibited long-lasting suppressive activity. Its intracellular localization without the membrane-permeable polyarginine sequence was possible via the formation of a stable -helix structure by stapling. Tumour bearing-mice treated daily or weekly with stapledERAP effectively prevented the BIG3-PHB2 interaction, leading to complete regression of E2-dependent tumours in vivo. Most importantly, combination of stapledERAP with tamoxifen, fulvestrant, and everolimus caused synergistic inhibitory effects on growth of breast cancer cells. Our findings suggested that the stapled ERAP may be a promising anti-tumour drug to suppress luminal-type breast cancer growth.
Yuki Shikata, Tetsuro Yoshimaru, Masato Komatsu, Hiroto Katoh, Reiko Sato, Shuhei Kanagaki, Yasumasa Okazaki, Shinya Toyokuni, Etsu Tashiro, Shumpei Ishikawa, Toyomasa Katagiri and Masaya Imoto : Protein kinase A inhibition facilitates the antitumor activity of xanthohumol, a valosin-containing protein inhibitor., Cancer Science, Vol.108, No.4, 785-794, 2017.
(要約)
Xanthohumol (XN), a simple prenylated chalcone, can be isolated from hops and has the potential to be a cancer chemopreventive agent against several human tumor cell lines. We previously identified valosin-containing protein (VCP) as a target of XN; VCP can also play crucial roles in cancer progression and prognosis. Therefore, we investigated the molecular mechanisms governing the contribution of VCP to the antitumor activity of XN. Several human tumor cell lines were treated with XN to investigate which human tumor cell lines are sensitive to XN. Several cell lines exhibited high sensitivity to XN both in vitro and in vivo. shRNA screening and bioinformatics analysis identified that the inhibition of the adenylate cyclase (AC) pathway synergistically facilitated apoptosis induced by VCP inhibition. These results suggest there is crosstalk between the AC pathway and VCP function, and targeting both VCP and the AC pathway is a potential chemotherapeutic strategy for a subset of tumor cells. This article is protected by copyright. All rights reserved.
Takayuki Iwamoto, Toyomasa Katagiri, Naoki Niikura, Yuichiro Miyoshi, Mariko Kochi, Tomohiro Nogami, Tadahiko Shien, Takayuki Motoki, Naruto Taira, Masako Omori, Yutaka Tokuda, Toshiyoshi Fujiwara, Hiroyoshi Doihara, Balazs Gyorffy and Junji Matsuoka : Immunohistochemical Ki67 after short-term hormone therapy identifies low-risk breast cancers as reliably as genomic markers., Oncotarget, Vol.8, No.16, 26122-26128, 2017.
(要約)
IHC based post-Ki67 levels may have distinct predictive power compared with the naïve IHC Ki67. Future studies with larger cohorts and longer follow-up periods may be needed to validate our results.
Yasuhiro Uno, Ryo Takata, Go Kito, Hiroshi Yamazaki, Kazuko Nakagawa, Yusuke Nakamura, Tetsuya Kamataki and Toyomasa Katagiri : Sex- and age-dependent gene expression in human liver: An implication for drug-metabolizing enzymes., Drug Metabolism and Pharmacokinetics, Vol.32, No.1, 100-107, 2016.
(要約)
Sex and age differences in hepatic expression of drug-metabolizing enzyme genes could cause variations in drug metabolism, but has not been fully elucidated, especially in Asian population. In this study, the global expression of human hepatic genes was analyzed by microarrays in 40 Japanese subjects (27 males and 13 females). Thirty-five sex-biased genes were identified (P < 0.005). Whereas, 60 age-biased genes in two age groups, <60 years and ≥70 years (P < 0.001), were identified in males. By Gene Ontology analysis, the sex-biased genes were related to protein catabolism and modification, while the age-biased genes were related to transcription regulation and cell death. Quantitative polymerase chain reaction confirmed the female-biased expression of drug-metabolizing enzyme genes BChE, CYP4X1, and SULT1E1 (≥1.5-fold, P < 0.05). Further analysis of drug-metabolizing enzyme genes indicated that expression of CYP2A6 and CYP3A4 in females in the ≥70 age group was less than in the <60 age group (≥1.5-fold, P < 0.05), and this trend was also observed for PXR expression in males (≥1.5-fold, P < 0.05). The results presented provide important insights into hepatic physiology and function, especially drug metabolism, with respect to sex and age.
Wataru Obara, Takashi Karashima, Kazuyoshi Takeda, Renpei Kato, Yoichiro Kato, Mitsugu Kanehira, Ryo Takata, Keiji Inoue, Toyomasa Katagiri, Taro Shuin, Yusuke Nakamura and Tomoaki Fujioka : Effective induction of cytotoxic T cells recognizing an epitope peptide derived from hypoxia-inducible protein 2 (HIG2) in patients with metastatic renal cell carcinoma., Cancer Immunology, Immunotherapy, Vol.66, No.1, 17-24, 2016.
(要約)
Vaccinations with HIG2-9-4 peptide could be well tolerated without any serious systemic adverse events. Peptide-specific cytotoxic T lymphocyte (CTL) responses were detected in eight of the nine patients. Doses of 1.0 or 3.0 mg/body seemed to induce a CTL response better than did a dose of 0.5 mg/body, although the number of patients was too small to draw a firm conclusion. The disease control rate (stable disease for ≥4 months) was 77.8 %, and the median progression-free survival time was 10.3 months.
Toru Nakamura, Toyomasa Katagiri, Shoki Sato, Toshihiro Kushibiki, Koji Hontani, Takahiro Tsuchikawa, Satoshi Hirano and Yusuke Nakamura : Overexpression of C16orf74 is involved in aggressive pancreatic cancers., Oncotarget, Vol.8, No.31, 50460-50475, 2016.
(要約)
Clinical outcome of pancreatic ductal adenocarcinoma (PDAC) has not been improved in the last three decades due to the lack of effective molecular-targeted drugs. To identify a novel therapeutic target for PDAC, we have performed genome-wide anamysis and found that Homo sapiens chromosome 16 open reading frame 74 (C16orf74) was up-regulated in the vast majority of PDAC. Overexpression of C16orf74protein detected by immunohistochemical analysis was an independent prognostic factor for patients with PDAC. The knockdown of endogenous C16orf74 expression in the PDAC cell lines KLM-1 and PK-59 by vector-based small hairpin-RNA (shRNA) drastically attenuated the growth of those cells, whereas ectopic C16orf74 overexpression in HEK293T and NIH3T3 cells promoted cell growth and invasion, respectively. More importantly, the endogenous threonine 44 (T44)-phosphorylated form of C16orf74 interacted with the protein phosphatase 3 catalytic subunit alpha (PPP3CA) via the PDIIIT sequence in the PPP3CA-binding motif within the middle portion of C16orf74 in PDAC cells. The overexpression of mutants of C16orf74 lacking the PDIIIT sequence or T44 phosphorylation resulted in the suppression of invasive activity compared with wild-type C16orf74, indicating that their interaction should be indispensable for PDAC cell invasion. These results suggest that C16orf74 plays an important role for PDAC invasion and proliferation, and is a promising target for a specific treatment for patients with PDAC.
Jae-Hyun Park, Miran Jang, Emre Yunus Tarhan, Toyomasa Katagiri, Mitsunori Sasa, Yasuo Miyoshi, R Krishna Kalari, J Vera Suman, Richard Weinshilboum, Liewei Wang, C Judy Boughey, P Matthew Goetz and Yusuke Nakamura : Clonal expansion of antitumor T cells in breast cancer correlates with response to neoadjuvant chemotherapy., International Journal of Oncology, Vol.49, No.2, 471-478, 2016.
(要約)
The immune microenvironment of tumor plays a critical role in therapeutic responses to chemotherapy. Cancer tissues are composed of a complex network between antitumor and pro-tumor immune cells and molecules; therefore a comprehensive analysis of the tumor immune condition is imperative for better understanding of the roles of the immune microenvironment in anticancer treatment response. In this study, we performed T cell receptor (TCR) repertoire analysis of tumor infiltrating T cells (TILs) in cancer tissues of pre- and post-neoadjuvant chemotherapy (NAC) from 19 breast cancer patients; five cases showed CR (complete response), ten showed PR (partial response), and four showed SD/PD (stable disease/progressive disease) to the treatment. From the TCR sequencing results, we calculated the diversity index of the TCRβ chain and found that clonal expansion of TILs could be detected in patients who showed CR or PR to NAC. Noteworthy, the diversity of TCR was further reduced in the post-NAC tumors of CR patients. Our quantitative RT-PCR also showed that expression ratio of CD8/Foxp3 was significantly elevated in the post-NAC tumors of CR cases (p=0.0032), indicating that antitumor T cells were activated and enriched in these tumors. Collectively, our findings suggest that the clonal expansion of antitumor T cells may be a critical factor associated with response to chemotherapy and that their TCR sequences might be applicable for the development of TCR-engineered T cells treatment for individual breast cancer patients when their tumors relapse.
Michiko Imamura, Arisa Nishimukai, Tomoko Higuchi, Hiromi Ozawa, Ayako Yanai, Yoshimasa Miyagawa, Keiko Murase, Isao Sakita, Takuya Hatada, Yuichi Takatsuka, Toyomasa Katagiri and Yasuo Miyoshi : High levels at baseline of serum pyridinoline crosslinked carboxyterminal telopeptide of type I collagen are associated with worse prognosis for breast cancer patients., Breast Cancer Research and Treatment, Vol.154, No.3, 521-531, 2015.
(要約)
It is speculated that adjuvant use of bisphosphonate reduces recurrence in breast cancer patients through suppression of bone resorption. To determine the prognostic impact of bone resorption markers, we investigated serum levels of the pyridinoline crosslinked carboxyterminal telopeptide of type I collagen (1CTP) and N-terminal crosslinking telopeptides of type I collagen (NTX). 1CTP and NTX were measured at baseline (before operation or neoadjuvant therapies) and afterward in 469 patients operated on breast cancer. The optimal cutoff value of 1CTP for relapse-free survival (RFS) was set at 3.6 ng/ml with an area under the receiver operating characteristics curve of 0.641 [95 % confidence interval (CI) = 0.560-0.721; p = 0.0011]. However, we were unable to determine a significant cutoff value for NTX. RFS was significantly worse for 1CTP-high patients with than for those with low levels of 1CTP (p = 0.0002). Multivariate analysis with tumor size, lymph node metastasis, and nuclear grade showed that 1CTP was a significant independent prognostic factor (hazard ratio = 2.04, 95 % CI = 1.13-3.68; p = 0.018). Worse prognosis for the subset with high 1CTP levels applied only to postmenopausal patients (p = 0.0002). RFS of 130 patients whose 1CTP changed from low at baseline to high at 6 months postoperatively showed RFS almost as poor as that for patients with high 1CTP throughout. These findings suggest that 1CTP may be useful not only for identifying patients with unfavorable prognosis, but also for selecting patients who may benefit from administration of bone-modifying agents in an adjuvant setting.
Namhee Kim, Tetsuro Yoshimaru, Yi-An Chen, Taisuke Matsuo, Masato Komatsu, Yasuo Miyoshi, Eiji Tanaka, Mitsunori Sasa, Kenji Mizuguchi and Toyomasa Katagiri : BIG3 Inhibits the Estrogen-Dependent Nuclear Translocation of PHB2 via Multiple Karyopherin-Alpha Proteins in Breast Cancer Cells., PLoS ONE, Vol.10, No.6, 2015.
(要約)
We recently reported that brefeldin A-inhibited guanine nucleotide-exchange protein 3 (BIG3) binds Prohibitin 2 (PHB2) in cytoplasm, thereby causing a loss of function of the PHB2 tumor suppressor in the nuclei of breast cancer cells. However, little is known regarding the mechanism by which BIG3 inhibits the nuclear translocation of PHB2 into breast cancer cells. Here, we report that BIG3 blocks the estrogen (E2)-dependent nuclear import of PHB2 via the karyopherin alpha (KPNA) family in breast cancer cells. We found that overexpressed PHB2 interacted with KPNA1, KPNA5, and KPNA6, thereby leading to the E2-dependent translocation of PHB2 into the nuclei of breast cancer cells. More importantly, knockdown of each endogenous KPNA by siRNA caused a significant inhibition of E2-dependent translocation of PHB2 in BIG3-depleted breast cancer cells, thereby enhancing activation of estrogen receptor alpha (ER). These data indicated that BIG3 may block the KPNAs (KPNA1, KPNA5, and KPNA6) binding region(s) of PHB2, thereby leading to inhibition of KPNAs-mediated PHB2 nuclear translocation in the presence of E2 in breast cancer cells. Understanding this regulation of PHB2 nuclear import may provide therapeutic strategies for controlling E2/ER signals in breast cancer cells.
Tetsuro Yoshimaru, Masato Komatsu, Yasuo Miyoshi, Junko Honda, Mitsunori Sasa and Toyomasa Katagiri : Therapeutic advances in BIG3-PHB2 inhibition targeting the crosstalk between estrogen and growth factors in breast cancer., Cancer Science, Vol.106, No.5, 550-558, 2015.
(要約)
Our previous studies demonstrated that specific inhibition of the BIG3-PHB2 complex, which is a critical modulator in estrogen (E2) signaling, using ERAP, a dominant negative peptide inhibitor, leads to suppression of E2-dependent estrogen receptor (ER) alpha activation through the reactivation of the tumor suppressive activity of PHB2. Here, we report that ERAP has significant suppressive effects against synergistic activation caused by the crosstalk between E2 and growth factors associated with intrinsic or acquired resistance to anti-estrogen tamoxifen in breast cancer cells. Intrinsic PHB2 released from BIG3 by ERAP effectively disrupted each interaction of membrane-associated ERα and insulin-like growth factor 1 receptor beta (IGF-1Rβ), EGFR, PI3K or human epidermal growth factor 2 (HER2) in the presence of E2 and the growth factors IGF or EGF, followed by inhibited the activation of IGF-1Rβ, EGFR or HER2, and reduced Akt, MAPK and ERα phosphorylation levels, resulting in significant suppression of proliferation of ERα-positive breast cancer cells in vitro and in vivo. More importantly, combined treatment with ERAP and tamoxifen led to a synergistic suppression of signaling that was activated by crosstalk between E2 and growth factors or HER2 amplification. Taken together, our findings suggest that the specific inhibition of BIG3-PHB2 is a novel potential therapeutic approach for the treatment of tamoxifen-resistant breast cancers activated by the crosstalk between E2 and growth factor signaling, especially in premenopausal women.
Ayako Yanai, Natsuko Inoue, Tomoko Yagi, Arisa Nishimukai, Yoshimasa Miyagawa, Keiko Murase, Michiko Imamura, Yukie Enomoto, Yuichi Takatsuka, Takahiro Watanabe, Seiichi Hirota, Mitsunori Sasa, Toyomasa Katagiri and Yasuo Miyoshi : Activation of mTOR/S6K But Not MAPK Pathways Might Be Associated With High Ki-67, ER(+), and HER2(-) Breast Cancer., Clinical Breast Cancer, Vol.15, No.3, 197-203, 2014.
(要約)
From our findings, we have concluded that the pS6 expression level is associated with the characteristics of breast cancer with high Ki-67 expression. Because these associations were observed, irrespective of menopausal status, the biologic difference seems to be less affected by estrogen signaling than by activation of S6 protein, especially in terms of proliferation. Our findings have also indicated that targeting the mTOR/S6K pathway might be a useful strategy for the treatment of ER(+)/HER2(-) breast cancer with high Ki-67 expression.
(キーワード)
Adult / Aged / Aged, 80 and over / Breast Neoplasms / Carcinoma, Ductal, Breast / Carcinoma, Lobular / Female / Humans / Ki-67 Antigen / Middle Aged / Mitogen-Activated Protein Kinases / Phosphatidylinositol 3-Kinases / Phosphorylation / Receptor, ErbB-2 / Receptors, Estrogen / Ribosomal Protein S6 Kinases / Signal Transduction / TOR Serine-Threonine Kinases
Tetsuro Yoshimaru, Masato Komatsu, Etsu Tashiro, Masaya Imoto, Hiroyuki Osada, Yasuo Miyoshi, Junko Honda, Mitsunoi Sasa and Toyomasa Katagiri : Xanthohumol suppresses oestrogen-signalling in breast cancer through the inhibition of BIG3-PHB2 interactions., Scientific Reports, Vol.4, 7355, 2014.
(要約)
Xanthohumol (XN) is a natural anticancer compound that inhibits the proliferation of oestrogen receptor- (ER)-positive breast cancer cells. However, the precise mechanism of the antitumour effects of XN on oestrogen (E2)-dependent cell growth, and especially its direct target molecule(s), remain(s) largely unknown. Here, we focus on whether XN directly binds to the tumour suppressor protein prohibitin 2 (PHB2), forming a novel natural antitumour compound targeting the BIG3-PHB2 complex and acting as a pivotal modulator of E2/ER signalling in breast cancer cells. XN treatment effectively prevented the BIG3-PHB2 interaction, thereby releasing PHB2 to directly bind to both nuclear- and cytoplasmic ER. This event led to the complete suppression of the E2-signalling pathways and ER-positive breast cancer cell growth both in vitro and in vivo, but did not suppress the growth of normal mammary epithelial cells. Our findings suggest that XN may be a promising natural compound to suppress the growth of luminal-type breast cancer.
Taisuke Matsuo, Le Tan Dat, Masato Komatsu, Tetsuro Yoshimaru, Kei Daizumoto, Saburo Sone, Yasuhiko Nishioka and Toyomasa Katagiri : Early growth response 4 is involved in cell proliferation of small cell lung cancer through transcriptional activation of its downstream genes., PLoS ONE, Vol.9, No.11, e113606, 2014.
(要約)
Small cell lung cancer (SCLC) is aggressive, with rapid growth and frequent bone metastasis; however, its detailed molecular mechanism remains poorly understood. Here, we report the critical role of early growth factor 4 (EGR4), a DNA-binding, zinc-finger transcription factor, in cell proliferation of SCLC. EGR4 overexpression in HEK293T cells conferred significant upregulation of specific splice variants of the parathyroid hormone-related protein (PTHrP) gene, resulting in enhancement of the secretion of PTHrP protein, a known mediator of osteolytic bone metastasis. More importantly, depletion of EGR4 expression by siRNA significantly suppressed growth of the SCLC cell lines, SBC-5, SBC-3 and NCI-H1048. On the other hand, introduction of EGR4 into NIH3T3 cells significantly enhanced cell growth. We identified four EGR4 target genes, SAMD5, RAB15, SYNPO and DLX5, which were the most significantly downregulated genes upon depletion of EGR4 expression in all of the SCLC cells examined, and demonstrated the direct recruitment of EGR4 to their promoters by ChIP and luciferase reporter analysis. Notably, knockdown of the expression of these genes by siRNA remarkably suppressed the growth of all the SCLC cells. Taken together, our findings suggest that EGR4 likely regulates the bone metastasis and proliferation of SCLC cells via transcriptional regulation of several target genes, and may therefore be a promising target for the development of anticancer drugs for SCLC patients.
Yi Paulisally Hau Lo, Chizu Tanikawa, Toyomasa Katagiri, Yusuke Nakamura and Koichi Matsuda : Identification of novel epigenetically inactivated gene PAMR1 in breast carcinoma., Oncology Reports, Vol.33, No.1, 267-273, 2014.
(要約)
Development of cancer is a complex process involving multiple genetic and epigenetic alterations. In our microarray analysis of 81 breast carcinoma specimens, we identified peptidase domain containing associated with muscle regeneration 1 (PAMR1) as being frequently suppressed in breast cancer tissues. PAMR1 expression was also reduced in all tested breast cancer cell lines, while PAMR1 was expressed moderately in normal breast tissues and primary mammary epithelial cells. DNA sequencing of the PAMR1 promoter after sodium bisulfite treatment revealed that CpG sites were hypermethylated in the breast cancer tissues and cell lines. PAMR1 expression was restored by 5-aza-2' deoxycytidine treatment, demonstrating that promoter hypermethylation contributed to PAMR1 inactivation in the breast cancer cells. In addition, ectopic expression of PAMR1 markedly suppressed cancer cell growth. In summary, our study identified PAMR1 as a putative tumor suppressor which was frequently inactivated by promoter hypermethylation in breast cancer tissues.
Yi-An Chen, Yoichi Murakami, Shandar Ahmad, Tetsuro Yoshimaru, Toyomasa Katagiri and Kenji Mizuguchi : Brefeldin A-inhibited guanine nucleotide-exchange protein 3 (BIG3) is predicted to interact with its partner through an ARM-type -helical structure., BMC Research Notes, Vol.7, No.1, 435, 2014.
(要約)
The combined results of the structure and interaction prediction led to a novel hypothesis that one of the predicted helices of BIG3 might play an important role in binding to PHB2 and thereby preventing its translocation to the nucleus. This hypothesis has been subsequently verified experimentally.
Sachiko Yoshimura, Takuya Tsunoda, Ryuji Osawa, Makiko Harada, Tomohisa Watanabe, Tetsuro Hikichi, Masahiro Katsuda, Motoki Miyazawa, Masaji Tani, Makoto Iwahashi, Kazuyoshi Takeda, Toyomasa Katagiri, Yusuke Nakamura and Hiroki Yamaue : Identification of an HLA-A2-restricted epitope peptide derived from hypoxia-inducible protein 2 (HIG2)., PLoS ONE, Vol.9, No.1, e85267, 2014.
(要約)
We herein report the identification of an HLA-A2 supertype-restricted epitope peptide derived from hypoxia-inducible protein 2 (HIG2), which is known to be a diagnostic marker and a potential therapeutic target for renal cell carcinoma. Among several candidate peptides predicted by the HLA-binding prediction algorithm, HIG2-9-4 peptide (VLNLYLLGV) was able to effectively induce peptide-specific cytotoxic T lymphocytes (CTLs). The established HIG2-9-4 peptide-specific CTL clone produced interferon- (IFN-) in response to HIG2-9-4 peptide-pulsed HLA-A*02:01-positive cells, as well as to cells in which HLA-A*02:01 and HIG2 were exogenously introduced. Moreover, the HIG2-9-4 peptide-specific CTL clone exerted cytotoxic activity against HIG2-expressing HLA-A*02:01-positive renal cancer cells, thus suggesting that the HIG2-9-4 peptide is naturally presented on HLA-A*02:01 of HIG-2-expressing cancer cells and is recognized by CTLs. Furthermore, we found that the HIG2-9-4 peptide could also induce CTLs under HLA-A*02:06 restriction. Taken together, these findings indicate that the HIG2-9-4 peptide is a novel HLA-A2 supertype-restricted epitope peptide that could be useful for peptide-based immunotherapy against cancer cells with HIG2 expression.
Taisuke Matsuo, Masato Komatsu, Tetsuro Yoshimaru, Kazuma Kiyotani, Yasuo Miyoshi, Mitsunori Sasa and Toyomasa Katagiri : Involvement of B3GALNT2 overexpression in the cell growth of breast cancer., International Journal of Oncology, Vol.44, No.2, 427-434, 2013.
(要約)
A number of glycosyltransferases have been identified and biologically characterized in cancer cells, yet their exact pathophysiological functions are largely unknown. Here, we report the critical role of 1,3-N-acetylgalactosaminyltransferase II (B3GALNT2), which transfers N-acetylgalactosamine (GalNAc) in a 1,3 linkage to N-acetylglucosamine, in the growth of breast cancer cells. Comprehensive transcriptomics, quantitative PCR and northern blot analyses indicated this molecule to be exclusively upregulated in the majority of breast cancers. Knockdown of B3GALNT2 expression by small interfering RNA attenuated cell growth and induced apoptosis in breast cancer cells. Overexpression of B3GALNT2 in HEK293T cells prompted secretion of the gene product into the culture medium, suggesting that B3GALNT2 is potentially a secreted protein. Furthermore, we demonstrated that B3GALNT2 is N-glycosylated on both Asn-116 and Asn-174 and that this modification is necessary for its secretion in breast cancer cells. Our findings suggest that this molecule represents a promising candidate for the development of a novel therapeutic targeting drug and a potential diagnostic tumor marker for patients with breast cancer, especially TNBC.
Siew-Kee Low, Atsushi Takahashi, Kyota Ashikawa, Johji Inazawa, Yoshio Miki, Michiaki Kubo, Yusuke Nakamura and Toyomasa Katagiri : Genome-wide association study of breast cancer in the Japanese population., PLoS ONE, Vol.8, No.10, 2013.
(要約)
Breast cancer is the most common malignancy among women in worldwide including Japan. Several studies have identified common genetic variants to be associated with the risk of breast cancer. Due to the complex linkage disequilibrium structure and various environmental exposures in different populations, it is essential to identify variants associated with breast cancer in each population, which subsequently facilitate the better understanding of mammary carcinogenesis. In this study, we conducted a genome-wide association study (GWAS) as well as whole-genome imputation with 2,642 cases and 2,099 unaffected female controls. We further examined 13 suggestive loci (P<1.0×10(-5)) using an independent sample set of 2,885 cases and 3,395 controls and successfully validated two previously-reported loci, rs2981578 (combined P-value of 1.31×10(-12), OR = 1.23; 95% CI = 1.16-.30) on chromosome 10q26 (FGFR2), rs3803662 (combined P-value of 2.79×10(-11), OR = 1.21; 95% CI = 1.15-.28) and rs12922061 (combined P-value of 3.97×10(-10), OR = 1.23; 95% CI = 1.15-.31) on chromosome 16q12 (TOX3-LOC643714). Weighted genetic risk score on the basis of three significantly associated variants and two previously reported breast cancer associated loci in East Asian population revealed that individuals who carry the most risk alleles in category 5 have 2.2 times higher risk of developing breast cancer in the Japanese population than those who carry the least risk alleles in reference category 1. Although we could not identify additional loci associated with breast cancer, our study utilized one of the largest sample sizes reported to date, and provided genetic status that represent the Japanese population. Further local and international collaborative study is essential to identify additional genetic variants that could lead to a better, accurate prediction for breast cancer.
Ayako Yanai, Yoshimasa Miyagawa, Keiko Murase, Michiko Imamura, Tomoko Yagi, Shigetoshi Ichii, Yuichi Takatsuka, Takashi Ito, Seiichi Hirota, Mitsunori Sasa, Toyomasa Katagiri and Yasuo Miyoshi : Influence of body mass index on clinicopathological factors including estrogen receptor, progesterone receptor, and Ki67 expression levels in breast cancers., International Journal of Clinical Oncology, Vol.19, No.3, 467-472, 2013.
(要約)
Premenopausal patients with high BMI showed a significantly higher frequency of lymph node metastasis (46.4 % vs. 22.9 %, P = 0.005) and tended to have a larger tumor size (P = 0.05) and higher nuclear grade (P = 0.07) than those with low BMI. These differences were not observed among postmenopausal patients. BMI was not associated with distribution of breast cancer subtypes, and ER, progesterone receptor (PR), and Ki67 expression levels of each subtype showed no differences between high and low BMI among premenopausal patients.
Masato Komatsu, Tetsuro Yoshimaru, Taisuke Matsuo, Kazuma Kiyotani, Yasuo Miyoshi, Toshihito Tanahashi, Kazuhito Rokutan, Rui Yamaguchi, Ayumu Saito, Seiya Imoto, Satoru Miyano, Yusuke Nakamura, Mitsunori Sasa, Mitsuo Shimada and Toyomasa Katagiri : Molecular features of triple negative breast cancer cells by genome-wide gene expression profiling analysis., International Journal of Oncology, Vol.42, No.2, 478-506, 2013.
(要約)
Triple negative breast cancer (TNBC) has a poor outcome due to the lack of beneficial therapeutic targets. To clarify the molecular mechanisms involved in the carcinogenesis of TNBC and to identify target molecules for novel anticancer drugs, we analyzed the gene expression profiles of 30 TNBCs as well as 13 normal epithelial ductal cells that were purified by laser-microbeam microdissection. We identified 301 and 321 transcripts that were significantly upregulated and downregulated in TNBC, respectively. In particular, gene expression profile analyses of normal human vital organs allowed us to identify 104 cancer-specific genes, including those involved in breast carcinogenesis such as NEK2, PBK and MELK. Moreover, gene annotation enrichment analysis revealed prominent gene subsets involved in the cell cycle, especially mitosis. Therefore, we focused on cell cycle regulators, asp (abnormal spindle) homolog, microcephaly-associated (Drosophila) (ASPM) and centromere protein K (CENPK) as novel therapeutic targets for TNBC. Small-interfering RNA-mediated knockdown of their expression significantly attenuated TNBC cell viability due to G1 and G2/M cell cycle arrest. Our data will provide a better understanding of the carcinogenesis of TNBC and could contribute to the development of molecular targets as a treatment for TNBC patients.
Yoichiro Kajita, Tomohisa Kato, Sakura Tamaki, Moritoshi Furu, Ryo Takahashi, Satoshi Nagayama, Tomoki Aoyama, Hiroyuki Nishiyama, Eijiro Nakamura, Toyomasa Katagiri, Yusuke Nakamura, Osamu Ogawa and Junya Toguchida : The transcription factor Sp3 regulates the expression of a metastasis-related marker of sarcoma, actin filament-associated protein 1-like 1 (AFAP1L1)., PLoS ONE, Vol.8, No.1, 2013.
(要約)
We previously identified actin filament-associated protein 1-like 1 (AFAP1L1) as a metastasis-predicting marker from the gene-expression profiles of 65 spindle cell sarcomas, and demonstrated the up-regulation of AFAP1L1 expression to be an independent risk factor for distant metastasis in multivariate analyses. Little is known, however, about how the expression of AFAP1L1 is regulated. Luciferase reporter assays showed tandem binding motives of a specificity protein (Sp) located at -85 to -75 relative to the transcriptional start site to be essential to the promoter activity. Overexpression of Sp1 and Sp3 proteins transactivated the proximal AFAP1L1 promoter construct, and electrophoretic mobility shift assays showed that both Sp1 and Sp3 were able to bind to this region in vitro. Chromatin immunoprecipitation experiments, however, revealed that Sp3 is the major factor binding to the proximal promoter region of the AFAP1L1 gene in AFAP1L1- positive cells. Treatment with mithramycin A, an inhibitor of proteins binding to GC-rich regions, prevented Sp3 from binding to the proximal promoter region of AFAP1L1 and decreased its expression in a dose-dependent manner. Finally, knocking down Sp3 using small inhibitory RNA duplex (siRNA) reduced AFAP1L1 expression significantly, which was partially restored by expressing siRNA-resistant Sp3. These findings indicate a novel role for Sp3 in sarcomas as a driver for expression of the metastasis-related gene AFAP1L1.
Tetsuro Yoshimaru, Masato Komatsu, Taisuke Matsuo, Yi-An Chen, Yoichi Murakami, Kenji Mizuguchi, Eiichi Mizohata, Tsuyoshi Inoue, Miki Akiyama, Rui Yamaguchi, Seiya Imoto, Satoru Miyano, Yasuo Miyoshi, Mitsunori Sasa, Yusuke Nakamura and Toyomasa Katagiri : Targeting BIG3-PHB2 interaction to overcome tamoxifen resistance in breast cancer cells., Nature Communications, Vol.4, 2443, 2013.
(要約)
The acquisition of endocrine resistance is a common obstacle in endocrine therapy of patients with oestrogen receptor- (ER)-positive breast tumours. We previously demonstrated that the BIG3-PHB2 complex has a crucial role in the modulation of oestrogen/ER signalling in breast cancer cells. Here we report a cell-permeable peptide inhibitor, called ERAP, that regulates multiple ER-signalling pathways associated with tamoxifen resistance in breast cancer cells by inhibiting the interaction between BIG3 and PHB2. Intrinsic PHB2 released from BIG3 by ERAP directly binds to both nuclear- and membrane-associated ER, which leads to the inhibition of multiple ER-signalling pathways, including genomic and non-genomic ER activation and ER phosphorylation, and the growth of ER-positive breast cancer cells both in vitro and in vivo. More importantly, ERAP treatment suppresses tamoxifen resistance and enhances tamoxifen responsiveness in ER-positive breast cancer cells. These findings suggest inhibiting the interaction between BIG3 and PHB2 may be a new therapeutic strategy for the treatment of luminal-type breast cancer.
Kazuma Kiyotani and Toyomasa Katagiri : A commentary on Analysis of ZNF350/ZBRK1 promoter variants and breast cancer susceptibility in non-BRCA1/2 French Canadian breast cancer families., Journal of Human Genetics, Vol.58, No.2, 58, 2012.
Tomoya Fukawa, Masaya Ono, Taisuke Matsuo, Hisanori Uehara, Tsuneharu Miki, Yusuke Nakamura, Hiro-omi Kanayama and Toyomasa Katagiri : DDX31 regulates the p53-HDM2 pathway and rRNA gene transcription through its interaction with NPM1 in renal cell carcinomas, Cancer Research, Vol.72, No.22, 5867-5877, 2012.
(要約)
Studies of renal cell carcinoma (RCC) have led to the development of new molecular-targeted drugs but its oncogenic origins remain poorly understood. Here, we report the identification and critical roles in renal carcinogenesis for DDX31, a novel nucleolar protein upregulated in the vast majority of human RCC. Immunohistochemical overexpression of DDX31 was an independent prognostic factor for patients with RCC. RNA interference (RNAi)-mediated attenuation of DDX31 in RCC cells significantly suppressed outgrowth, whereas ectopic DDX31 overexpression in human 293 kidney cells drove their proliferation. Endogenous DDX31 interacted and colocalized with nucleophosmin (NPM1) in the nucleoli of RCC cells, and attenuation of DDX31 or NPM1 expression decreased pre-ribosomal RNA biogenesis. Notably, in DDX31-attenuated cells, NPM1 was translocated from nucleoli to the nucleoplasm or cytoplasm where it bound to HDM2. As a result, HDM2 binding to p53 was reduced, causing p53 stablization with concomitant G(1) phase cell-cycle arrest and apoptosis. Taken together, our findings define a mechanism through which control of the DDX31-NPM1 complex is likely to play critical roles in renal carcinogenesis.
Seham Elgazzar, Hitoshi Zembutsu, Atsushi Takahashi, Michiaki Kubo, Fuminori Aki, Koichi Hirata, Yuichi Takatsuka, Minoru Okazaki, Shozo Ohsumi, Takashi Yamakawa, Mitsunori Sasa, Toyomasa Katagiri, Yoshio Miki and Yusuke Nakamura : A genome-wide association study identifies a genetic variant in the SIAH2 locus associated with hormonal receptor-positive breast cancer in Japanese., Journal of Human Genetics, Vol.57, No.12, 766-771, 2012.
(要約)
In Japan, breast cancer is the most common cancer among women and the second leading cause of cancer death among women worldwide. To identify genetic variants associated with the disease susceptibility, we performed a genome-wide association study (GWAS) using a total of 1086 Japanese female patients with hormonal receptor-positive (HRP) breast cancer and 1816 female controls. We selected 33 single-nucleotide polymorphisms (SNPs) with suggestive associations in GWAS (P-value of <1 × 10(-4)) as well as 4 SNPs that were previously implicated their association with breast cancer for further replication by an independent set of 1653 cases and 2797 controls. We identified significant association of the disease with a SNP rs6788895 (P(combined) of 9.43 × 10(-8) with odds ratio (OR) of 1.22) in the SIAH2 (intron of seven in absentia homolog 2) gene on chromosome 3q25.1 where the involvement in estrogen-dependent diseases was suggested. In addition, rs3750817 in intron 2 of the fibroblast growth factor receptor 2 gene, which was reported to be associated with breast cancer susceptibility, was significantly replicated with P(combined) of 8.47 × 10(-8) with OR=1.22. Our results suggest a novel susceptibility locus on chromosome 3q25.1 for a HRP breast cancer.
Keiko Murase, Ayako Yanai, Masaru Saito, Michiko Imamura, Yoshimasa Miyagawa, Yuichi Takatsuka, Natsuko Inoue, Takashi Ito, Seiichi Hirota, Mitsunori Sasa, Toyomasa Katagiri, Yasuhisa Fujimoto, Takuya Hatada, Shigetoshi Ichii, Tomoyuki Nishizaki, Naohiro Tomita and Yasuo Miyoshi : Biological characteristics of luminal subtypes in pre- and postmenopausal estrogen receptor-positive and HER2-negative breast cancers., Breast Cancer, Vol.21, No.1, 52-57, 2012.
(要約)
BACKGROUND: Estrogen receptor (ER)-positive and human epidermal growth factor receptor 2 (HER2)-negetive breast cancers can be divided into luminal A and luminal B subtypes based on Ki67 expression levels. However, the biological differences in ER and progesterone receptor (PR) expression levels between these luminal subtypes are not clear. METHODS: We examined immunohistochemical expression levels of ER, PR, and Ki67 in 180 ER-positive/HER2-negative breast cancers while taking menopausal status into account. Breast cancers were divided according to ER and PR levels (H: >50%, L: ≤50%), and luminal A and B were classified by the Ki67 labeling index (A: Ki67 <14%, B: Ki67 ≥14%). RESULTS: When breast cancers were classified based on ER and PR levels, the distribution of pre- and postmenopausals was significantly different for luminal A (P < 0.0001), but not for luminal B cancers. As for luminal A, ER-H/PR-L cancers were rare among premenopausals (8%), but frequent among postmenopausals (54%). Correlation between ER and PR levels among luminal A cancers was strong in premenopausals but weak in postmenopausals. Since crosstalk with growth factor signaling is unlikely in luminal A, we speculate that intratumoral estrogen insufficiency contributed to the characteristics of postmenopausal ER-H/PR-L cancers. CONCLUSION: We speculate that the biological characteristics of luminal A cancers are influenced by the estrogen environment, but its influence on luminal B cancers may be limited. We believe these considerations constitute useful information for a better understanding of the biology of ER-positive-HER2-negetive breast cancers.
Le T Dat, Taisuke Matsuo, Tetsuro Yoshimaru, Souji Kakiuchi, Hisatsugu Goto, Masaki Hanibuchi, Takuya Kuramoto, Yasuhiko Nishioka, Saburo Sone and Toyomasa Katagiri : Identification of genes potentially involved in bone metastasis by genome-wide gene expression profile analysis of non-small cell lung cancer in mice, International Journal of Oncology, Vol.40, No.5, 1455-1469, 2012.
(要約)
Lung cancer is commonly associated with multi-organ metastasis, and the bone is a frequent metastatic site for lung cancer. However, the molecular mechanism of organ-specific metastasis remains poorly understood. To elucidate this issue, we analyzed in this study genome-wide gene expression profiles of 15 metastatic lesions from three organs (bone, lung and liver) in a mouse model with multi-organ metastasis properties of human non-small cell lung cancer cells (ACC-LC319/bone2), using a combination of laser-microbeam microdissection and DNA microarrays. We identified 299 genes that could potentially be involved in the organ-selective nature of lung cancer metastasis. Among them, 77 were bone-specifically expressed elements, including genes involved in cell adhesion, cytoskeleton/cell motility, extracellular matrix remodeling and cell-cell signaling as well as genes already known to be involved in the bone metastasis of breast cancers. Quantitative RT-PCR confirmed the specific upregulation of eight genes in bone metastasis tumors, suggesting that these genes may be involved in bone metastasis. Our findings should be helpful for a better understanding of the molecular aspects of the metastatic process in different organs, and could lead to molecular target-based anticancer drugs and prevention of metastasis, especially bone metastasis.
Hiroki Yoshioka, Shinji Yamamoto, Hirofumi Hanaoka, Yasuhiko Iida, Pramila Paudyal, Tetsuya Higuchi, Hideyuki Tominaga, Noboru Oriuchi, Hidewaki Nakagawa, Yasuhiro Shiba, Koji Yoshida, Ryuji Osawa, Toyomasa Katagiri, Takuya Tsunoda, Yusuke Nakamura and Keigo Endo : In vivo therapeutic effect of CDH3/P-cadherin-targeting radioimmunotherapy., Cancer Immunology, Immunotherapy, Vol.61, No.8, 1211-1220, 2012.
(要約)
Our in vivo results confirmed the specific binding of MAb-6 to tumor cells after intravenous injections of (111)In-labeled MAb-6 to mice with tumors expressing CDH3/P-cadherin. A single intravenous injection of (90)Y-MAb-6 (100 μCi) significantly suppressed tumor growth in mice with tumors expressing CDH3/P-cadherin. Furthermore, two injections of (90)Y-MAb-6 led to complete tumor regression in H1373-inoculated mice without any detectable toxicity.
Takeaki Kawai, M Jose M Caaveiro, Ryota Abe, Toyomasa Katagiri and Kouhei Tsumoto : Catalytic activity of MsbA reconstituted in nanodisc particles is modulated by remote interactions with the bilayer., FEBS Letters, Vol.585, No.22, 3533-3537, 2011.
(要約)
ATP-binding cassette (ABC) transporters couple hydrolysis of ATP with vectorial transport across the cell membrane. We have reconstituted ABC transporter MsbA in nanodiscs of various sizes and lipid compositions to test whether ATPase activity is modulated by the properties of the bilayer. ATP hydrolysis rates, Michaelis-Menten parameters, and dissociation constants of substrate analog ATP--S demonstrated that physicochemical properties of the bilayer modulated binding and ATPase activity. This is remarkable when considering that the catalytic unit is located ~50Å from the transmembrane region. Our results validated the use of nanodiscs as an effective tool to reconstitute MsbA in an active catalytic state, and highlighted the close relationship between otherwise distant transmembrane and ATPase modules.
M Furu, Y Kajita, S Nagayama, T Ishibe, Y Shima, K Nishijo, D Uejima, R Takahashi, T Aoyama, T Nakayama, T Nakamura, Y Nakashima, M Ikegawa, S Imoto, Toyomasa Katagiri, Y Nakamura and J Toguchida : Identification of AFAP1L1 as a prognostic marker for spindle cell sarcomas., Oncogene, Vol.30, No.38, 4015-4025, 2011.
(要約)
Spindle cell sarcomas consist of tumors with different biological features, of which distant metastasis is the most ominous sign for a poor prognosis. However, metastasis is difficult to predict on the basis of current histopathological analyses. We have identified actin filament-associated protein 1-like 1 (AFAP1L1) as a candidate for a metastasis-predicting marker from the gene expression profiles of 65 spindle cell sarcomas. A multivariate analysis determined that AFAP1L1 was an independent factor for predicting the occurrence of distant metastasis (P=0.0001), which was further confirmed in another set of 41 tumors by a quantitative mRNA expression analysis. Immunohistochemical staining using paraffin-embedded tumor tissues revealed that the metastasis-free rate was significantly better in tumors negative for AFAP1L1 (P=0.0093 by log-rank test). Knocking down the AFAP1L1 gene in sarcoma cells resulted in inhibition of the cell invasion, and forced expression of AFAP1L1 in immortalized human mesenchymal stem cells induced anchorage-independent growth and increased cell invasiveness with high activity levels of matrix metallopeptidase. Furthermore, tumor growth in vivo was accelerated in AFAP1L1-transduced sarcoma cell lines. These results suggest that AFAP1L1 has a role in the progression of spindle cell sarcomas and is a prognostic biomarker.
Jae-Hyun Park, Toyomasa Katagiri, Suyoun Chung, Kyoko Kijima and Yusuke Nakamura : Polypeptide N-acetylgalactosaminyltransferase 6 disrupts mammary acinar morphogenesis through O-glycosylation of fibronectin., Neoplasia, Vol.13, No.4, 320-326, 2011.
(要約)
A high expression of short and immature O-glycans is one of the prominent features of breast cancer cells, which would be attributed to the upregulated expression of glycosyltransferases. Therefore, a detailed elucidation of glycosyltransferases and their substrate(s) may improve our understandings for their roles in mammary carcinogenesis. Here we report that overexpression of polypeptide N-acetylgalactosaminyltransferase 6 (GALNT6), a glycosyltransferase involved in the initial step of O-glycosylation, has transformational potentials through disruptive acinar morphogenesis and cellular changes similar to epithelial-to-mesenchymal transition in normal mammary epithelial cell, MCF10A. As one of the critical O-glycan substrates, we identified fibronectin that was O-glycosylated in vivo and thereby stabilized by GALNT6. Because knockdown of fibronectin abrogated the disruptive proliferation caused by introduction of GALNT6 into epithelial cells, our findings suggest that GALNT6-fibronectin pathway should be a critical component for breast cancer development and progression.
(キーワード)
Breast Neoplasms / Carcinoma / Cell Line / Disease Progression / Female / Fibronectins / Gene Knockdown Techniques / Glycosylation / Humans / Isoenzymes / Mammary Glands, Human / Models, Biological / Morphogenesis / N-Acetylgalactosaminyltransferases / Protein Stability / Transfection
Masahiko Ajiro, Toshihiko Nishidate, Toyomasa Katagiri and Yusuke Nakamura : Critical involvement of RQCD1 in the EGFR-Akt pathway in mammary carcinogenesis., International Journal of Oncology, Vol.37, No.5, 1085-1093, 2010.
(要約)
We previously reported an important role of RQCD1 in mammary carcinogenesis through the interaction with Grb10 interacting GYF protein 1 (GIGYF1), Grb10 interacting GYF protein 2 (GIGYF2) and growth factor receptor binding protein 10 (Grb10). In this study, we investigated the biological mechanism of RQCD1 in regulation of the Akt activity as the downstream signal of epidermal growth factor receptor (EGFR). Knockdown of RQCD1 reduced the Akt phosphorylation level that was induced by epidermal growth factor (EGF) stimulation. We found a possible formation of the big complex involved in the Akt activity including Akt, EGFR, GIGYF1 and GIGYF2, Grb10 and RQCD1. We subsequently defined that a region corresponding to 620-665th amino acids of GIGYF1 and 667-712th amino acids of GIGYF2 interacted with RQCD1. Furthermore, we found that RQCD1 was required for enhancement of the interaction of Grb10 with GIGYF1 and GIGYF2. Our findings in this study imply the functional mechanism of RQCD1 in the Akt activity regulation as a mediator in the EGFR-signaling pathway.
Chikako Fukukawa, Koji Ueda, Toshihiko Nishidate, Toyomasa Katagiri and Yusuke Nakamura : Critical roles of LGN/GPSM2 phosphorylation by PBK/TOPK in cell division of breast cancer cells., Genes, Chromosomes & Cancer, Vol.49, No.10, 861-872, 2010.
(要約)
To investigate the molecular mechanism of mammary carcinogenesis and identify novel molecular targets for breast cancer therapy, we analyzed genome-wide gene expression profiles of 81 clinical breast cancer samples. Here, we report the critical role of LGN/GPSM2 (Leu-Gly-Asn repeat-enriched protein/G-protein signaling modulator 2) in the growth of breast cancer cells. Semiquantitative RT-PCR and Northern blot analyses confirmed upregulation of LGN/GPSM2 in a large proportion of breast cancers. Immunocytochemical staining identified LGN/GPSM2 at the spindle in cells at metaphase, and at midzone and midbody in cytokinetic cells. Western blot analysis indicated the highest expression and the phosphorylated form of LGN/GPSM2 protein in G2/M phase. Treatment with small-interfering RNAs (siRNAs) targeting LGN/GPSM2 caused incompletion of cell division and resulted in significant growth suppression of breast cancer cells. We found that the 450th threonine (Thr450) of LGN/GPSM2 was phosphorylated by the serine/threonine kinase PBK/TOPK during mitosis. Overexpression of LGN/GPSM2-T450A in which Thr450 was substituted with alanine induced growth suppression and aberrant chromosomal segregation. These findings imply an important role of LGN/GPSM2 in cell division of breast cancer cells and suggest that the PBK/TOPK-LGN/GPSM2 pathway might be a promising molecular target for treatment of breast cancer.
Jung-Won Kim, Chikako Fukukawa, Koji Ueda, Toshihiko Nishidate, Toyomasa Katagiri and Yusuke Nakamura : Involvement of C12orf32 overexpression in breast carcinogenesis., International Journal of Oncology, Vol.37, No.4, 861-867, 2010.
(要約)
Through genome-wide gene expression profile analysis of breast cancer, we identified a gene, chromosome 12 open reading frame 32 (C12orf32), to be involved in mammary carcinogenesis. Semiquantitative RT-PCR and Northern blot analysis confirmed C12orf32 overexpression in breast cancer cells and its almost undetectable level of expression in normal human tissues. Immunocytochemical staining analysis using breast cancer cell lines revealed a cell cycle-dependent subcellular localization of endogenous C12orf32 protein. Depletion of C12orf32 expression by small-hairpin RNA interference significantly suppressed the growth of breast cancer cell lines possibly due to the inhibition of G1/S transition and subsequent cell death. Western blot analysis indicated that a C12orf32 protein of 35 kDa predicted from the cDNA sequences was processed to a 16-kDa protein of (C12orf32-p16) which was accumulated in most of breast cancer cell lines examined. Our data suggest that C12orf32 is a promising molecular target for the development of novel anticancer drugs such as peptide vaccines and siRNA drugs.
Yosuke Harada, Mitsugu Kanehira, Yoshiko Fujisawa, Ryo Takata, Taro Shuin, Tsuneharu Miki, Tomoaki Fujioka, Yusuke Nakamura and Toyomasa Katagiri : Cell-permeable peptide DEPDC1-ZNF224 interferes with transcriptional repression and oncogenicity in bladder cancer cells., Cancer Research, Vol.70, No.14, 5829-5839, 2010.
(要約)
Bladder cancer is the second most common genitourinary cancer worldwide, yet its oncogenic origins remain poorly understood. The cancer-testis antigen DEPDC1 was shown recently to contribute to bladder cancer oncogenesis. In this study, we examined the biological functions of DEPDC1 and defined a potential therapeutic strategy to target this molecule. Coimmunoprecipitation and immunocytochemistry revealed that DEPDC1 interacted and colocalized with zinc finger transcription factor ZNF224, a known transcriptional repressor. Inhibiting this interaction with a cell-permeable peptide corresponding to the ZNF224-interacting domain in DEPDC1 induced apoptosis of bladder cancer cells in vitro and in vivo. By inhibiting DEPDC1-ZNF224 complex formation, this peptide triggered transcriptional activation of A20, a potent inhibitor of the NF-kappaB signaling pathway. Our findings indicate that the DEPDC1-ZNF224 complex is likely to play a critical role in bladder carcinogenesis.
Jae-Hyun Park, Toshihiko Nishidate, Kyoko Kijima, Takao Ohashi, Kaoru Takegawa, Tomoko Fujikane, Koichi Hirata, Yusuke Nakamura and Toyomasa Katagiri : Critical roles of mucin 1 glycosylation by transactivated polypeptide N-acetylgalactosaminyltransferase 6 in mammary carcinogenesis., Cancer Research, Vol.70, No.7, 2759-2769, 2010.
(要約)
The structure of O-glycosylated proteins is altered in breast cancer cells, but the mechanisms of such an aberrant modification have been largely unknown. We here report critical roles of a novel druggable target, polypeptide N-acetylgalactosaminyltransferase 6 (GALNT6), which is upregulated in a great majority of breast cancers and encodes a glycosyltransferase responsible for initiating mucin-type O-glycosylation. Knockdown of GALNT6 by small interfering RNA significantly enhanced cell adhesion function and suppressed the growth of breast cancer cells. Western blot and immunostaining analyses indicated that wild-type GALNT6 protein could glycosylate and stabilize an oncoprotein mucin 1 (MUC1), which was upregulated with GALNT6 in breast cancer specimens. Furthermore, knockdown of GALNT6 or MUC1 led to similar morphologic changes of cancer cells accompanied by the increase of cell adhesion molecules beta-catenin and E-cadherin. Our findings implied that overexpression of GALNT6 might contribute to mammary carcinogenesis through aberrant glycosylation and stabilization of MUC1 and that screening of GALNT6 inhibitors would be valuable for the development of novel therapeutic modalities against breast cancer.
Tomomi Ueki, Jae-Hyun Park, Toshihiko Nishidate, Kyoko Kijima, Koichi Hirata, Yusuke Nakamura and Toyomasa Katagiri : Ubiquitination and downregulation of BRCA1 by ubiquitin-conjugating enzyme E2T overexpression in human breast cancer cells., Cancer Research, Vol.69, No.22, 8752-8760, 2009.
(要約)
Breast cancer is generated through a multistep genetic and epigenetic process including activations of oncogenes and inactivations of tumor suppressor genes. Here, we report a critical role of ubiquitin-conjugating enzyme E2T (UBE2T), an E2 ubiquitin-conjugating enzyme, in mammary carcinogenesis. Immunocytochemical staining and in vitro binding assay revealed that UBE2T interacted and colocalized with the BRCA1/BRCA1-associated RING domain protein (BARD1) complex. Knocking down of UBE2T expression with small interfering RNA drastically suppressed the growth of breast cancer cells. Interestingly, in vivo ubiquitination assay indicated BRCA1 to be polyubiquitinated by incubation with wild-type UBE2T protein, but not with C86A-UBE2T protein, an E2 activity-dead mutant, in which the 86th residue of cysteine was replaced with alanine. Furthermore, knocking down of UBE2T protein induced upregulation of BRCA1 protein in breast cancer cells, whereas its overexpression caused the decrease of the BRCA1 protein. Our data imply a critical role of UBE2T in development and/or progression of breast cancer through the interaction with and the regulation of the BRCA1/BARD1 complex.
Jae-Hyun Park, Toshihiko Nishidate, Yusuke Nakamura and Toyomasa Katagiri : Critical roles of T-LAK cell-originated protein kinase in cytokinesis., Cancer Science, Vol.101, No.2, 403-411, 2009.
(要約)
We previously reported up-regulation of T-LAK cell-originated protein kinase (TOPK), a novel mitotic kinase, in the great majority of breast cancers. Here we report its critical roles in mitosis, especially in cytokinesis. We found that protein phosphatase 1 alpha (PP1alpha) inactivation by cyclin-dependent kinase 1 (CDK1)/cyclin B1 caused enhancement of autophosphorylation of TOPK and resulted in its activation at an early stage of mitosis. Then TOPK interacted with and phosphorylated p97, a member of the AAA+ family of ATPase proteins, through an interaction with p47 protein as an adaptor protein. Interestingly, knockdown of TOPK or p97 in breast cancer cells caused the mitotic failures in the abscission process. This mitotic failure could be rescued by additional exogenous introduction of wild-type TOPK protein, but not by that of its kinase-dead form. Our findings suggest that TOPK is indispensable for cancer cell cytokinesis throughout phosphorylation on p97.
Masahiko Ajiro, Toyomasa Katagiri, Koji Ueda, Hidewaki Nakagawa, Chikako Fukukawa, Meng-Lay Lin, Jae-Hyun Park, Toshihiko Nishidate, Yataro Daigo and Yusuke Nakamura : Involvement of RQCD1 overexpression, a novel cancer-testis antigen, in the Akt pathway in breast cancer cells., International Journal of Oncology, Vol.35, No.4, 673-681, 2009.
(要約)
We here report identification and characterization of required for cell differentiation 1 homolog (RQCD1) as a potential therapeutic target for breast cancer. Gene-expression profiling analysis of breast cancer cells, semi-quantitative RT-PCR, Northern blotting and Western blotting confirmed RQCD1 to be frequently up-regulated in breast cancer specimens and breast cancer cell lines. On the other hand, its expression was very weak or hardly detectable in normal human tissues except testis, indicating this molecule to be a novel cancer-testis antigen. Treatment of breast cancer cell lines with siRNA targeting RQCD1 drastically suppressed cell proliferation. Concordantly, introduction of exogenous RQCD1 into HEK293 cells significantly enhanced cell growth, implying RQCD1 to have an oncogenic activity. Co-immunoprecipitation experiments and immunocytochemical staining revealed an interaction of RQCD1 protein with Grb10 interacting GYF protein 1 (GIGYF1) and 2 (GIGYF2) proteins, involved in regulation of Akt activation, in breast cancer cells. Interestingly, knockdown of either of RQCD1, GIGYF1 or GIGYF2 resulted in significant reduction of the phosphorylation of Akt at Ser 473 in breast cancer cell lines. Our findings suggest that RQCD1 is a potential molecular target for treatment of breast cancer.
Jung-Won Kim, Miki Akiyama, Jae-Hyun Park, Meng-Lay Lin, Arata Shimo, Tomomi Ueki, Yataro Daigo, Tatsuhiko Tsunoda, Toshihiko Nishidate, Yusuke Nakamura and Toyomasa Katagiri : Activation of an estrogen/estrogen receptor signaling by BIG3 through its inhibitory effect on nuclear transport of PHB2/REA in breast cancer., Cancer Science, Vol.100, No.8, 1468-1478, 2009.
(要約)
Breast cancer is known to be a hormone-dependent disease, and estrogens through an interaction with estrogen receptor (ER) enhance the proliferative and metastatic activity of breast tumor cells. Here we show a critical role of transactivation of BIG3, brefeldin A-inhibited guanine nucleotide-exchange protein 3, in activation of the estrogen/ER signaling in breast cancer cells. Knocking-down of BIG3 expression with small-interfering RNA (siRNA) drastically suppressed the growth of breast cancer cells. Subsequent coimmunoprecipitation and immunoblotting assays revealed an interaction of BIG3 with prohibitin 2/repressor of estrogen receptor activity (PHB2/REA). When BIG3 was absent, stimulation of estradiol caused the translocation of PHB2/REA to the nucleus, enhanced the interaction of PHB2/REA and ERalpha, and resulted in suppression of the ERalpha transcriptional activity. On the other hand, when BIG3 was present, BIG3 trapped PHB2/REA in the cytoplasm and inhibited its nuclear translocation, and caused enhancement of ERalpha transcriptional activity. Our results imply that BIG3 overexpression is one of the important mechanisms causing the activation of the estrogen/ERalpha signaling pathway in the hormone-related growth of breast cancer cells.
Meng-Lay Lin, Chikako Fukukawa, Jae-Hyun Park, Kie Naito, Kyoko Kijima, Arata Shimo, Masahiko Ajiro, Toshihiko Nishidate, Yusuke Nakamura and Toyomasa Katagiri : Involvement of G-patch domain containing 2 overexpression in breast carcinogenesis., Cancer Science, Vol.100, No.8, 1443-1450, 2009.
(要約)
Through analysis of the detailed genome-wide gene expression profiles of 81 breast tumors, we identified a novel gene, G-patch domain containing 2 (GPATCH2), that was overexpressed in the great majority of breast cancer cases. Treatment of breast cancer cells MCF-7 and T47D with siRNA against GPATCH2 effectively suppressed its expression, and resulted in the growth suppression of cancer cells, suggesting its essential role in breast cancer cell growth. We found an interaction of GPATCH2 protein with hPrp43, an RNA-dependent ATPase. Their interaction could significantly enhance the ATPase activity of hPrp43 and induce a growth-promoting effect on mammalian cells. Because northern blot analyses of normal human organs implied GPATCH2 to be a novel cancer/testis antigen, targeting GPATCH2 or inhibition of the interaction between GPATCH2 and hPrp43 could be a promising novel therapeutic strategy of breast cancer.
Hirofumi Hanaoka, Toyomasa Katagiri, Chikako Fukukawa, Hiroki Yoshioka, Shinji Yamamoto, Yasuhiko Iida, Tetsuya Higuchi, Noboru Oriuchi, Bishnuhari Paudyal, Pramila Paudyal, Yusuke Nakamura and Keigo Endo : Radioimmunotherapy of solid tumors targeting a cell-surface protein, FZD10: therapeutic efficacy largely depends on radiosensitivity., Annals of Nuclear Medicine, Vol.23, No.5, 479-485, 2009.
(要約)
OBJECTIVE: Frizzled homolog 10 (FZD10) is expressed at high levels on the cell surface of almost all synovial sarcoma tissues, but is absent in most normal organs. In a previous study, yttrium-90 ((90)Y)-labeled anti-FZD10 antibody (MAb 92-13) showed considerable therapeutic efficacy in synovial sarcoma cell-bearing mice. The purpose of the present study was to elucidate the factors associated with this therapeutic efficacy of (90)Y-MAb 92-13. METHODS: FZD10 expression levels of SYO-1 (FZD10-overexpressing synovial sarcoma cell line) and DLD-1/FZD10 (FZD10-transfected DLD-1 cell) were determined by the cell binding assay, and their radiosensitivity was evaluated by incubation with (90)Y-MAb 92-13 in vitro. Biodistribution study of indium-111 ((111)In)-MAb 92-13 was performed in SYO-1 and DLD-1/FZD10 tumor-bearing mice. For therapeutic studies, SYO-1 and DLD-1/FZD10 tumor-bearing mice were treated with (90)Y-MAb 92-13 (100, 150, and 200 muCi), after which the change in tumor volume was measured. Immunohistochemical staining was performed on the excised tumor. RESULTS: Expression level of FZD10 on DLD-1/FZD10 was much greater than that on SYO-1. The accumulation of (111)In-MAb 92-13 was much higher in DLD-1/FZD10 tumor-bearing mice than in SYO-1 tumor-bearing mice (49.0 +/- 4.2 and 22.0 +/- 4.5% ID/g, respectively, at 48 h after administration). In SYO-1 tumor, substantial tumor size reduction was observed in all mice treated with (90)Y-MAb 92-13 (tumor volume decreased to less than 0.1 cm(3) at 11 days after treatment) and tumor regrowth was not observed in most of them. In contrast, only slow progression was observed in DLD-1/FZD10 tumor. When incubated with (90)Y-MAb 92-13, high radioactivity was needed to damage DLD-1/FZD10. Immunohistochemical study indicated apoptosis of SYO-1 tumor. CONCLUSIONS: The therapeutic efficacy of RIT seems to largely depend on the tumor radiosensitivity.
Osteosarcoma is the most prevalent bone malignant tumor in children and adolescents, and displays heterogeneous histology and high propensity for distant metastasis. Although adjuvant chemotherapy remarkably improved treatment outcome over the past few decades, prognosis for osteosarcoma patients with pulmonary metastasis is still unsatisfactory. To identify novel therapeutic targets for osteosarcoma, we investigated the gene expression profile of osteosarcomas by cDNA microarray analysis and found transactivation of receptor tyrosine kinase-like orphan receptor 2 (ROR2) expression in the majority of osteosarcoma samples. Treatment of osteosarcoma cell lines with siRNA against ROR2 significantly inhibited cell proliferation and migration. We also identified wingless-type MMTV integration site family, member 5B (WNT5B) as a putative ROR2 ligand and that the physiological interaction of WNT5B and ROR2 could enhance cell migration, indicating the possible roles of ROR2 and WNT5B in the metastatic property of osteosarcoma cells. Taken together, our findings revealed that the WNT5B/ROR2 signaling pathway is a promising therapeutic target for osteosarcoma.
Koji Ueda, Yu Fukase, Toyomasa Katagiri, Nobuhisa Ishikawa, Shinji Irie, Taka-Aki Sato, Hiroyuki Ito, Haruhiko Nakayama, Yohei Miyagi, Eiju Tsuchiya, Nobuoki Kohno, Mieko Shiwa, Yusuke Nakamura and Yataro Daigo : Targeted serum glycoproteomics for the discovery of lung cancer-associated glycosylation disorders using lectin-coupled ProteinChip arrays., Proteomics, Vol.9, No.8, 2182-2192, 2009.
(要約)
To screen for glycoproteins showing aberrant sialylation patterns in sera of cancer patients and apply such information for biomarker identification, we performed SELDI-TOF MS analysis coupled with lectin-coupled ProteinChip arrays (Jacalin or SNA) using sera obtained from lung cancer patients and control individuals. Our approach consisted of three processes (i) removal of 14 abundant proteins in serum, (ii) enrichment of glycoproteins with lectin-coupled ProteinChip arrays, and (iii) SELDI-TOF MS analysis with acidic glycoprotein-compatible matrix. We identified 41 protein peaks showing significant differences (p<0.05) in the peak levels between the cancer and control groups using the Jacalin- and SNA-ProteinChips. Among them, we identified loss of Neu5Ac (alpha2,6) Gal/GalNAc structure in apolipoprotein C-III (apoC-III) in cancer patients through subsequent MALDI-QIT-TOF MS/MS. Furthermore, subsequent validation experiments using an additional set of 60 lung adenocarcinoma patients and 30 normal controls demonstrated that there is a higher frequency of serum apoC-III with loss of alpha2,6-linkage Neu5Ac residues in lung cancer patients compared to controls. Our results have demonstrated that lectin-coupled ProteinChip technology allows the high-throughput and specific recognition of cancer-associated aberrant glycosylations, and implied a possibility of its applicability to studies on other diseases.
C Fukukawa, Satoshi Nagayama, T Tsunoda, J Toguchida, Yusuke Nakamura and Toyomasa Katagiri : Activation of the non-canonical Dvl-Rac1-JNK pathway by Frizzled homologue 10 in human synovial sarcoma., Oncogene, Vol.28, No.8, 1110-20, 2009.
(要約)
We previously reported that Frizzled homologue 10 (FZD10), a member of the Wnt signal receptor family, was highly and specifically upregulated in synovial sarcoma and played critical roles in its cell survival and growth. We here report a possible molecular mechanism of the FZD10 signaling in synovial sarcoma cells. We found a significant enhancement of phosphorylation of the Dishevelled (Dvl)2/Dvl3 complex as well as activation of the Rac1-JNK cascade in synovial sarcoma cells in which FZD10 was overexpressed. Activation of the FZD10-Dvls-Rac1 pathway induced lamellipodia formation and enhanced anchorage-independent cell growth cells. FZD10 overexpression also caused the destruction of the actin cytoskeleton structure, probably through the downregulation of the RhoA activity. Our results have strongly implied that FZD10 transactivation causes the activation of the non-canonical Dvl-Rac1-JNK pathway and plays critical roles in the development/progression of synovial sarcomas.
S Dobashi, Toyomasa Katagiri, Eiji Hirota, Shingo Ashida, Y Daigo, T Shuin, T Fujioka, Tuneharu Miki and Yusuke Nakamura : Involvement of TMEM22 overexpression in the growth of renal cell carcinoma cells., Oncology Reports, Vol.21, No.2, 305-312, 2009.
(要約)
In order to clarify the molecular mechanism involved in renal carcinogenesis, and to identify molecular targets for development of novel treatments of renal cell carcinoma (RCC), we previously analyzed genome-wide gene expression profiles of clear-cell types of RCC by cDNA microarray. Among the transcativated genes, we herein focused on functional significance of TMEM22 (transmembrane protein 22), a transmembrane protein, in cell growth of RCC. Northern blot and semi-quantitative RT-PCR analyses confirmed up-regulation of TMEM22 in a great majority of RCC clinical samples and cell lines examined. Immunocytochemical analysis validated its localization at the plasma membrane. We found an interaction between TMEM22 and RAB37 (Ras-related protein Rab-37), which was also up-regulated in RCC cells. Interestingly, knockdown of either of TMEM22 or RAB37 expression by specific siRNA caused significant reduction of cancer cell growth. Our results imply that the TMEM22/RAB37 complex is likely to play a crucial role in growth of RCC and that inhibition of the TMEM22/RAB37 expression or their interaction should be novel therapeutic targets for RCC.
Satoshi Nagayama, Eiji Yamada, Yoshiki Kohno, Tomoki Aoyama, Chikako Fukukawa, Hajime Kubo, Go Watanabe, Toyomasa Katagiri, Yusuke Nakamura, Yoshiharu Sakai and Junya Toguchida : Inverse correlation of the up-regulation of FZD10 expression and the activation of beta-catenin in synchronous colorectal tumors., Cancer Science, Vol.100, No.3, 405-412, 2008.
(要約)
We investigated the immunohistochemical expression patterns of Frizzled homolog 10 (FZD10), a cell-surface receptor for molecules in the Wnt pathway, in tissue samples derived from 104 patients with colorectal cancers (CRCs). There was no immunoreactivity for FZD10 in normal colonic mucosa, and only tumor cells in polyps and CRC tissues showed spotted immunostaining patterns in apical sides of the cytoplasm. In metastatic liver lesions, tumor cells showed cytoplasmic immunostaining similar to primary lesions, whereas normal liver parenchyma showed almost no immunostaining. Frequencies of FZD10-immunopositive cells in tumor tissues were significantly higher in CRCs than those in polyps (3.3 +/- 10.3% vs 20.5 +/- 31.7%, P = 0.0016), and almost equivalent with those in metastatic liver lesions (33.2 +/- 39.7% vs 26.4 +/- 33.4%, P = 0.133). Analyses of paired samples (polyps and CRCs, or CRCs and metastatic liver lesions from the same patient) suggested that a subset of CRCs possessed intrinsic genetic mechanisms causing the evolution of FZD10-positive clones during tumor progression, making FZD10 a promising candidate for molecular imaging and a target for therapy. To our surprise, cancer cells immunopositive for FZD10 showed significantly less nuclear accumulation of beta-catenin, compared to FZD10-immunonegative cancer cells, and there was a strong inverse correlation between nuclear immunostaining scores for beta-catenin expression and expression patterns of FZD10 (P = 0.0002), suggesting that FZD10 has a distinct role from other FZDs in canonical Wnt signal transduction.
Hiroyuki Inoue, Mutsunori Iga, Hayuka Nabeta, Tomoko Yokoo, Yoko Suehiro, Shinji Okano, Makoto Inoue, Hiroaki Kinoh, Toyomasa Katagiri, Koichi Takayama, Yoshikazu Yonemitsu, Mamoru Hasegawa, Yusuke Nakamura, Yoichi Nakanishi and Kenzaburo Tani : Non-transmissible Sendai virus encoding granulocyte macrophage colony-stimulating factor is a novel and potent vector system for producing autologous tumor vaccines., Cancer Science, Vol.99, No.11, 2315-2326, 2008.
(要約)
The recent clinical application of granulocyte macrophage colony-stimulating factor (GM-CSF)-transduced autologous tumor vaccines revealed substantial antitumor activity and valuable clinical results. However, for these vaccines to be optimally effective, the antitumor efficacies must be improved. Recently, Sendai virus (SeV) vectors, which are cytoplasmic RNA vectors, have emerged as safe vectors with high gene transduction. In the current study, the in vivo therapeutic antitumor efficacies of irradiated GM-CSF-transduced mouse renal cell carcinoma (RENCA) vaccine cells mediated by either fusion gene-deleted non-transmissible SeV encoding mouse GM-CSF (SeV/dF/G) or adenovirus (E1, E3 deleted serotype 5 adenovirus) encoding mouse GM-CSF (AdV/G) (respectively described as irRC/SeV/GM or irRC/AdV/GM) were compared in RENCA-bearing mice. The results showed that the antitumor effect was equivalent between irRC/SeV/GM and irRC/AdV/GM cells, even though the former produced less GM-CSF in vitro. The cell numbers of activated (CD80(+), CD86(+), CD80( (+) )CD86(+)) dendritic cells in lymph nodes from mice treated with irRC/AdV/GM or irRC/SeV/GM cells were increased significantly compared with those of mice treated with the respective controls, at both the earlier and later phases. In an in vitro cytotoxicity assay, splenocytes harvested from mice treated with both irRC/SeV/GM and irRC/AdV/GM cells showed tumor-specific responses against RENCA cells. The restimulated splenocytes harvested from mice treated with irRC/SeV/GM or irRC/AdV/GM cells produced significantly higher levels of interleukin-2, interleukin-4, and interferon-gamma compared with their respective controls (P < 0.05). Furthermore, vaccination with irRC/AdV/GM or irRC/SeV/GM cells induced significantly enhanced recruitment of the cytolytic effectors of CD107a(+)CD8(+) T cells and CD107a(+) natural killer cells into tumors compared with those induced by their respective controls (P < 0.05). Taken together, our results suggest that the SeV/dF/G vector is a potential candidate for the production of effective autologous GM-CSF-transduced tumor vaccines in clinical cancer immune gene therapy.
T Ueki, T Nishidate, JH Park, ML Lin, Arata Shimo, Koichi Hirata, Yusuke Nakamura and Toyomasa Katagiri : Involvement of elevated expression of multiple cell-cycle regulator, DTL/RAMP (denticleless/RA-regulated nuclear matrix associated protein), in the growth of breast cancer cells., Oncogene, Vol.27, No.43, 5672-83, 2008.
(要約)
To investigate the detailed molecular mechanism of mammary carcinogenesis and discover novel therapeutic targets, we previously analysed gene expression profiles of breast cancers. We here report characterization of a significant role of DTL/RAMP (denticleless/RA-regulated nuclear matrix associated protein) in mammary carcinogenesis. Semiquantitative RT-PCR and northern blot analyses confirmed upregulation of DTL/RAMP in the majority of breast cancer cases and all of breast cancer cell lines examined. Immunocytochemical and western blot analyses using anti-DTL/RAMP polyclonal antibody revealed cell-cycle-dependent localization of endogenous DTL/RAMP protein in breast cancer cells; nuclear localization was observed in cells at interphase and the protein was concentrated at the contractile ring in cytokinesis process. The expression level of DTL/RAMP protein became highest at G(1)/S phases, whereas its phosphorylation level was enhanced during mitotic phase. Treatment of breast cancer cells, T47D and HBC4, with small-interfering RNAs against DTL/RAMP effectively suppressed its expression and caused accumulation of G(2)/M cells, resulting in growth inhibition of cancer cells. We further demonstrate the in vitro phosphorylation of DTL/RAMP through an interaction with the mitotic kinase, Aurora kinase-B (AURKB). Interestingly, depletion of AURKB expression with siRNA in breast cancer cells reduced the phosphorylation of DTL/RAMP and decreased the stability of DTL/RAMP protein. These findings imply important roles of DTL/RAMP in growth of breast cancer cells and suggest that DTL/RAMP might be a promising molecular target for treatment of breast cancer.
R Konda, J Sugimura, F Sohma, Toyomasa Katagiri, Yusuke Nakamura and T Fujioka : Over expression of hypoxia-inducible protein 2, hypoxia-inducible factor-1alpha and nuclear factor kappaB is putatively involved in acquired renal cyst formation and subsequent tumor transformation in patients with end stage renal failure., The Journal of Urology, Vol.180, No.2, 481-485, 2008.
(要約)
We examined hypoxia-inducible protein 2, hypoxia-inducible factor-1alpha and nuclear factor-kappaB in acquired cystic disease of the kidney associated with renal cell carcinoma to elucidate the roles of these factors in cyst formation and subsequent tumor transformation. Immunohistochemical expression of hypoxia-inducible protein 2, hypoxia-inducible factor-1alpha and phosphorylated nuclear factor-kappaB (active form) were examined in 20 normal kidney samples obtained from nephrectomy for localized renal cell carcinoma and 25 kidneys with acquired cystic disease associated renal cell carcinoma from 23 patients on dialysis. Only faint or weak immunostaining for hypoxia-inducible protein 2, hypoxia-inducible factor-1alpha and phosphorylated nuclear factor-kappaB was observed in normal kidney tissues. In nontumor areas of the kidneys with acquired cystic disease expressions of these 3 proteins was up-regulated in tubular and cyst epithelial cells. Acquired cysts were classified into 3 types according to cyst epithelium morphology, namely flat, cuboidal and hyperplastic. Hyperplastic cysts were the predominant cysts expressing hypoxia-inducible protein 2 and hypoxia-inducible factor-1alpha. Although up-regulation of hypoxia-inducible protein 2, hypoxia-inducible factor-1alpha and phosphorylated nuclear factor-kappaB was observed in renal cell carcinoma, positive hypoxia-inducible protein 2 immunostaining was detected predominantly in papillary renal cell carcinoma, while positive hypoxia-inducible factor-1alpha and phosphorylated nuclear factor-kappaB immunostaining was prominent in clear cell renal cell carcinoma. Hypoxia-inducible protein 2, hypoxia-inducible factor-1alpha and phosphorylated nuclear factor-kappaB may be involved in a continuous process of the evolution of phenotypic expression from a simple cyst to epithelial hyperplasia and eventually to tumor.
T Kidokoro, C Tanikawa, Y Furukawa, Toyomasa Katagiri, Yusuke Nakamura and Koichi Matsuda : CDC20, a potential cancer therapeutic target, is negatively regulated by p53., Oncogene, Vol.27, No.11, 1562-71, 2008.
(要約)
The p53 protein inhibits malignant transformation through direct and indirect regulation of transcription of many genes related to cell cycle, apoptosis and cellular senescence. A number of genes induced by p53 have been well characterized, but biological significance of genes whose expression was suppressed by p53 is still largely undisclosed. To clarify the roles of p53-suppressive genes in carcinogenesis, we analysed two data sets of whole-genome expression profiles, one for cells in which wild-type p53 was exogenously introduced and the other for a large number of clinical cancer tissues. Here, we identified CDC20 that was frequently upregulated in many types of malignancies and remarkably suppressed by ectopic introduction of p53. CDC20 expression was suppressed by genotoxic stresses in p53- and p21-dependent manners through CDE-CHR elements in the CDC20 promoter. Furthermore, small interference RNA (siRNA)-mediated silencing of p53 induced CDC20 expression in normal human dermal fibroblast cells. As we expected, treatment of cancer cells with siRNA against CDC20 induced G(2)/M arrest and suppressed cell growth. Our results indicate that p53 inhibits tumor cell growth through the indirect regulation of CDC20 and that CDC20 might be a good potential therapeutic target for a broad spectrum of human cancer.
C Fukukawa, Hirofumi Hanaoka, Satoshi Nagayama, T Tsunoda, J Toguchida, Keigo Endo, Yusuke Nakamura and Toyomasa Katagiri : Radioimmunotherapy of human synovial sarcoma using a monoclonal antibody against FZD10., Cancer Science, Vol.99, No.2, 432-440, 2008.
(要約)
We previously reported Frizzled homolog 10 (FZD10), a member of the Frizzled family, to be a promising therapeutic target for synovial sarcomas. In this report, we established a murine monoclonal antibody (MAb), namely, MAb 92-13 that had specific binding activity against native FZD10 product expressed in synovial sarcoma cell lines. Subsequent immunohistochemical analyses with the MAb 92-13 confirmed an absence or hardly detectable level of FZD10 protein in any normal human organs. We confirmed the specific binding activity of this MAb in vivo after injection of fluorescent-labeled MAb i.p. or i.v. into the mice carrying synovial sarcoma xenografts by the use of the in vivo fluorescent imaging system as well as radioisotopes. Moreover, MAb 92-13 was effectively internalized into the synovial sarcoma cells after its binding to FZD10 on the cell surface. A single i.v. injection of the Yttrium-90 ((90)Y)-MAb 92-13 drastically suppressed tumor growth of synovial sarcoma in mice without any severe toxicity. Median time to tumor progression was 58 days for mice treated with (90)Y-MAb 92-13 and 9 days for mice treated with non-labeled antibody control or untreated mice (difference = 49 days; P = 7 x 10(-5)). This result indicates that MAb 92-13 could be utilized as the novel treatment modality for synovial sarcoma and other FZD10-positive tumors.
Arata Shimo, Chizu Tanikawa, Toshihiko Nishidate, ML Lin, Koichi Matsuda, JH Park, Tomomi Ueki, Tomohiko Ohta, Koichi Hirata, Mamoru Fukuda, Yusuke Nakamura and Toyomasa Katagiri : Involvement of kinesin family member 2C/mitotic centromere-associated kinesin overexpression in mammary carcinogenesis., Cancer Science, Vol.99, No.1, 62-70, 2008.
(要約)
To elucidate the molecular mechanisms of mammary carcinogenesis and discover novel therapeutic targets for breast cancer, we previously carried out genome-wide expression profile analysis of 81 breast cancer cases by means of cDNA microarray coupled with laser microbeam microdissection of cancer cells. Among the dozens of transactivated genes, in the present study we focused on the functional significance of kinesin family member 2C (KIF2C)/mitotic centromere-associated kinesin (MCAK) in the growth of breast cancer cells. Northern blot and immunohistochemical analyses confirmed KIF2C/MCAK overexpression in breast cancer cells, and showed that it is expressed at undetectable levels in normal human tissues except the testis, suggesting KIF2C/MCAK to be a cancer-testis antigen. Western blot analysis using breast cancer cell lines revealed a significant increase in the endogenous KIF2C/MCAK protein level and its phosphorylation in G(2)/M phase. Treatment of breast cancer cells with small interfering RNA against KIF2C/MCAK effectively suppressed KIF2C/MCAK expression and inhibited the growth of the breast cancer cell lines T47D and HBC5. In addition, we found that KIF2C/MCAK expression was significantly suppressed by ectopic introduction of p53. These findings suggest that overexpression of KIF2C/MCAK might be involved in breast carcinogenesis and is a promising therapeutic target for breast cancers.
Yoichiro Kamatani, Koichi Matsuda, Tetsuya Ohishi, Shigeru Ohtsubo, Keiko Yamazaki, Aritoshi Iida, Naoya Hosono, Michiaki Kubo, Wako Yumura, Kosaku Nitta, Toyomasa Katagiri, Yasushi Kawaguchi, Naoyuki Kamatani and Yusuke Nakamura : Identification of a significant association of a single nucleotide polymorphism in TNXB with systemic lupus erythematosus in a Japanese population., Journal of Human Genetics, Vol.53, No.1, 64-73, 2008.
(要約)
Systemic lupus erythematosus (SLE) is one of the common autoimmune diseases, with complex genetic components. Here, we report on a case-control association study of 178 SLE patients and 899 control subjects, using genome-wide gene-based single nucleotide polymorphism (SNP) markers. An SNP, rs3130342, in a 5' flanking region of the TNXB gene revealed a significant association with SLE [P = 0.000000930, odds ratio (OR) 3.11, with 95% confidence interval (95%CI) of 1.89-5.28] in a Japanese population. This association was replicated independently with 203 cases and 294 controls (P = 0.0440, OR 1.52, with 95%CI of 1.01-2.78). Although a copy number variation (CNV) of the C4 gene adjacent to the TNXB gene was reported to be associated with SLE, our analysis on this CNV revealed that the association of CNV of the C4 gene was weaker than the SNP in the TNXB gene and likely to reflect the linkage disequilibrium between C4 CNV and this particular SNP. Stratified analysis also revealed that the association of SNP rs3130342 with SLE was independent of the HLA-DRB1*1501 allele that has been shown to be associated with SLE. Our findings strongly imply that the TNXB gene is a candidate gene susceptible to SLE in the Japanese population.
(キーワード)
Chromosomes, Human, Pair 6 / Gene Frequency / Genetics, Population / Genotype / Humans / Lupus Erythematosus, Systemic / Polymorphism, Single Nucleotide / Tenascin
K Ueda, Toyomasa Katagiri, T Shimada, S Irie, T Sato, Y Nakamura and Y Daigo : Comparative profiling of serum glycoproteome by sequential purification of glycoproteins and 2-nitrobenzensulfenyl (NBS) stable isotope labeling: a new approach for the novel biomarker discovery for cancer, Journal of Proteome Research, Vol.6, No.9, 3475-83, 2007.
(要約)
The recent progress in various proteomic technologies allows us to screen serum biomarker including carbohydrate antigens. However, only a limited number of proteins could be detected by current conventional methods such as shotgun proteomics, primarily because of the enormous concentration distribution of serum proteins and peptides. To circumvent this difficulty and isolate potential cancer-specific biomarkers for diagnosis and treatment, we established a new screening system consisting of the sequential steps of (1) immunodepletion of 6 high-abundance proteins, (2) targeted enrichment of glycoproteins by lectin column chromatography, and (3) the quantitative proteome analysis using 12C6- or 13C6-NBS (2-nitrobenzenesulfenyl) stable isotope labeling followed by MALDI-QIT-TOF mass spectrometric analysis. Through this systematic analysis for five serum samples derived from patients with lung adenocarcinoma, we identified as candidate biomarkers 34 serum glycoproteins that revealed significant difference in alpha1,6-fucosylation level between lung cancer and healthy control, clearly demonstrating that the carbohydrate-focused proteomics could allow for the detection of serum components with cancer-specific features. In addition, we developed a more simplified and practical technique, mass spectrometry-based glycan structure analysis and lectin blotting, in order to validate glycan structure of candidate biomarkers that could be applicable in clinical use. Our new glycoproteomic strategy will provide highly sensitive and quantitative profiling of specific glycan structures on multiple proteins, which should be useful for serum biomarker discovery.
S Senju, H Suemori, H Zembutsu, Y Uemura, S Hirata, D Fukuma, H Matsuyoshi, M Shimomura, M Haruta, S Fukushima, Y Matsunaga, Toyomasa Katagiri, Y Nakamura, M Furuya, N Nakatsuji and Y Nishimura : Genetically manipulated human embryonic stem cell-derived dendritic cells with immune regulatory function, Stem Cells, Vol.25, No.11, 2720-2729, 2007.
(要約)
Genetically manipulated dendritic cells (DC) are considered to be a promising means for antigen-specific immune therapy. This study reports the generation, characterization, and genetic modification of DC derived from human embryonic stem (ES) cells. The human ES cell-derived DC (ES-DC) expressed surface molecules typically expressed by DC and had the capacities to stimulate allogeneic T lymphocytes and to process and present protein antigen in the context of histocompatibility leukocyte antigen (HLA) class II molecule. Genetic modification of human ES-DC can be accomplished without the use of viral vectors, by the introduction of expression vector plasmids into undifferentiated ES cells by electroporation and subsequent induction of differentiation of the transfectant ES cell clones to ES-DC. ES-DC introduced with invariant chain-based antigen-presenting vectors by this procedure stimulated HLA-DR-restricted antigen-specific T cells in the absence of exogenous antigen. Forced expression of programmed death-1-ligand-1 in ES-DC resulted in the reduction of the proliferative response of allogeneic T cells cocultured with the ES-DC. Generation and genetic modification of ES-DC from nonhuman primate (cynomolgus monkey) ES cells was also achieved by the currently established method. ES-DC technology is therefore considered to be a novel means for immune therapy.
H Zembutsu, M Yanada, A Hishida, Toyomasa Katagiri, T Tsuruo, I Sugiura, J Takeuchi, N Usui, T Naoe, Y Nakamura and R Ohno : Prediction of risk of disease recurrence by genome-wide cDNA microarray analysis in patients with Philadelphia chromosome-positive acute lymphoblastic leukemia treated with imatinib-combined chemotherapy, International Journal of Oncology, Vol.31, No.2, 313-322, 2007.
(要約)
Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ALL) reveals very poor prognosis due to high incidence of relapse when treated with standard chemotherapy. Although >96% of patients with Ph+ALL achieved complete remission (CR) with imatinib-combined chemotherapy in a phase II clinical trial conducted by the Japan Adult Leukemia Study Group (JALSG), 26% of them experienced hematological relapse in a short time after achievement of CR. In this study, to establish a prediction system for risk of relapse, we analyzed gene expression profiles of 23 bone marrow samples from patients with Ph+ALL using cDNA microarray consisting of 27,648 cDNA sequences. Using the 19 randomly-selected test cases, we identified 16 genes that were expressed significantly differently between patients with (n=8) and without (n=11) continuous response; from the list of 16 genes, we selected the 6 'predictive' genes and constructed a numerical scoring system by which the two groups were clearly separated, with positive scores for the former and the negative scores for the latter. Scoring of 4 cases that were reserved from the original 23 cases predicted correctly their clinical responses. In addition, three cases whose BCR-Abl transcript levels failed to reduce sufficiently after induction therapy, also revealed negative scores. We also developed a quantitative reverse transcription-PCR-based prediction system that could be feasible for routine clinical use. Our results suggest that achievement of continuous response with imatinib-combined chemotherapy can be predicted by expression patterns in this set of genes, leading to achievement of 'personalized therapy' for treatment of this disease.
M Fujikawa, Toyomasa Katagiri, A Tugores, Y Nakamura and F Ishikawa : ESE-3, an Ets family transcription factor, is up-regulated in cellular senescence, Cancer Science, Vol.98, No.9, 1468-1475, 2007.
(要約)
Normal cells irreversibly stop dividing after being exposed to a variety of stresses. This state, called cellular senescence, has recently been demonstrated to act as a tumor-suppressing mechanism in vivo. A common set of features are exhibited by senescent cells, but the molecular mechanism leading to the state is poorly understood. It has been shown that p38, a stress-induced mitogen-activated protein kinase (MAPK), plays a pivotal role in inducing cellular senescence in diverse settings. To better understand the senescence-inducing pathway, microarray analyses of normal human fibroblasts that ectopically activated p38 were performed. It was found that five genes encoding ESE-3, inhibin betaA, RGS5, SSAT and DIO2 were up-regulated in senescent cells induced by RasV12, H(2)O(2) and telomere shortening, but not in quiescent or actively growing cells, suggesting that these genes serve as molecular markers for various types of cellular senescence. The ectopic expression of ESE-3 resulted in retarded growth, up-regulation of p16(INK4a) but not of p21, and increased levels of SA-beta-gal activity. In contrast, RGS5, SSAT and the constitutive active form of the inhibin betaA receptor gene did not induce such senescence phenotypes when ectopically expressed. ESE-3 expression increased the activity of the p16(INK4a) promoter in a reporter assay, and recombinant ESE-3 protein bound to the Ets-binding sequences present in the promoter. These results suggest that ESE-3 plays a role in the induction of cellular senescence as a downstream molecule of p38.
M Kanehira, Y Harada, R Takata, T Shuin, T Miki, T Fujioka, Y Nakamura and Toyomasa Katagiri : Involvement of upregulation of DEPDC1 (DEP domain containing 1) in bladder carcinogenesis, Oncogene, Vol.26, No.44, 6448-55, 2007.
(要約)
In an attempt to disclose mechanisms of bladder carcinogenesis and discover novel target molecules for development of treatment, we applied a cDNA microarray to screen genes that were significantly transactivated in bladder cancer cells. Among the upregulated genes, we here focused on a novel gene, (DEPDC1) DEP domain containing 1, whose overexpression was confirmed by northern blot and immunohistochemical analyses. Immunocytochemical staining analysis detected strong staining of endogenous DEPDC1 protein in the nucleus of bladder cancer cells. Since DEPDC1 expression was hardly detectable in any of 24 normal human tissues we examined except the testis, we considered this gene-product to be a novel cancer/testis antigen. Suppression of DEPDC1 expression with small-interfering RNA significantly inhibited growth of bladder cancer cells. Taken together, these findings suggest that DEPDC1 might play an essential role in the growth of bladder cancer cells, and would be a promising molecular-target for novel therapeutic drugs or cancer peptide-vaccine to bladder cancers.
M Kanehira, Toyomasa Katagiri, A Shimo, R Takata, T Shuin, T Miki, T Fujioka and Y Nakamura : Oncogenic role of MPHOSPH1, a cancer-testis antigen specific to human bladder cancer, Cancer Research, Vol.67, No.7, 3276-85, 2007.
(要約)
To disclose the molecular mechanism of bladder cancer, the second most common genitourinary tumor, we had previously done genome-wide expression profile analysis of 26 bladder cancers by means of cDNA microarray representing 27,648 genes. Among genes that were significantly up-regulated in the majority of bladder cancers, we here report identification of M-phase phosphoprotein 1 (MPHOSPH1) as a candidate molecule for drug development for bladder cancer. Northern blot analyses using mRNAs of normal human organs and cancer cell lines indicated this molecule to be a novel cancer-testis antigen. Introduction of MPHOSPH1 into NIH3T3 cells significantly enhanced cell growth at in vitro and in vivo conditions. We subsequently found an interaction between MPHOSPH1 and protein regulator of cytokinesis 1 (PRC1), which was also up-regulated in bladder cancer cells. Immunocytochemical analysis revealed colocalization of endogenous MPHOSPH1 and PRC1 proteins in bladder cancer cells. Interestingly, knockdown of either MPHOSPH1 or PRC1 expression with specific small interfering RNAs caused a significant increase of multinuclear cells and subsequent cell death of bladder cancer cells. Our results imply that the MPHOSPH1/PRC1 complex is likely to play a crucial role in bladder carcinogenesis and that inhibition of the MPHOSPH1/PRC1 expression or their interaction should be novel therapeutic targets for bladder cancers.
A Shimo, T Nishidate, T Ohta, M Fukuda, Y Nakamura and Toyomasa Katagiri : Elevated expression of PRC1, protein regulator of cytokinesis 1, involved in the growth of breast cancer cells, Cancer Science, Vol.98, No.2, 174-181, 2007.
(要約)
To elucidate molecular mechanisms of mammary carcinogenesis and discover novel therapeutic targets for breast cancer, we previously carried out a genome-wide expression profile analysis of 81 breast cancer cases by means of a combination of cDNA microarray and laser microbeam microdissection. Among the upregulated genes, we focused on the functional significance of protein regulator of cytokinesis 1 (PRC1) in the development of breast cancer. Western blot analysis using breast cancer cell lines revealed a significant increase in endogenous PRC1 levels in G(2)/M phase. Treatment of breast cancer cells with small interfering RNA against PRC1 effectively suppressed its expression and inhibited the growth of breast cancer cell lines T47D and HBC5. Furthermore, we found an interaction between PRC1 and kinesin family member 2C/mitotic centromere-associated kinesin (KIF2C/MCAK) by coimmunoprecipitation and immunoblotting using COS-7 cells, in which these molecules were introduced exogenously. These findings suggest the involvement of a PRC1-KIF2C/MCAK complex in breast tumorigenesis, and this complex should be a promising target for the development of novel treatments for breast cancer.
(キーワード)
Animals / Breast Neoplasms / Cell Cycle / Cell Cycle Proteins / Cell Line / Cercopithecus aethiops / Gene Expression Regulation, Neoplastic / Genetic Variation / Humans / Kinesin / Protein Binding / RNA, Small Interfering
Ryo Takata, Toyomasa Katagiri, Mitsugu Kanehira, Taro Shuin, Tsuneharu Miki, Mikio Namiki, Kenjiro Kohri, Tatsuhiko Tsunoda, Tomoaki Fujioka and Yusuke Nakamura : Validation study of the prediction system for clinical response of M-VAC neoadjuvant chemotherapy., Cancer Science, Vol.98, No.1, 113-117, 2007.
(要約)
To predict the efficacy of the M-VAC neoadjuvant chemotherapy for invasive bladder cancers, we previously established the method to calculate the prediction score on the basis of expression profiles of 14 predictive genes. This scoring system had clearly distinguished the responder group from the non-responder group. To further validate the clinical significance of the system, we applied it to 22 additional cases of bladder cancer patients and found that the scoring system correctly predicted clinical response for 19 of the 22 test cases. The group of patients with positive predictive scores had significantly longer survival than that with negative scores. When we compared our results with a previous report describing the prognosis of the patients with cystectomy alone, the results imply that patients with positive scores are likely to benefit from M-VAC neoadjuvant chemotherapy, but that the chemotherapy would shorten the lives of patients with negative scores. We are confident that our prediction system to M-VAC therapy should provide opportunities for achieving better prognosis and improving the quality of life of patients. Taken together, our data suggest that the goal of 'personalized medicine', prescribing the appropriate treatment regimen for each patient, may be achievable by selecting specific sets of genes for their predictive values according to the approach shown here.
ML Lin, JH Park, T Nishidate, Y Nakamura and Toyomasa Katagiri : Involvement of maternal embryonic leucine zipper kinase (MELK) in mammary carcinogenesis through interaction with Bcl-G, a pro-apoptotic member of the Bcl-2 family., Breast Cancer Research, Vol.9, No.1, R17, 2007.
(要約)
Cancer therapies directed at specific molecular targets in signaling pathways of cancer cells, such as tamoxifen, aromatase inhibitors and trastuzumab, have proven useful for treatment of advanced breast cancers. However, increased risk of endometrial cancer with long-term tamoxifen administration and of bone fracture due to osteoporosis in postmenopausal women undergoing aromatase inhibitor therapy are recognized side effects. These side effects as well as drug resistance make it necessary to search for novel molecular targets for drugs on the basis of well-characterized mechanisms of action. Using accurate genome-wide expression profiles of breast cancers, we found maternal embryonic leucine-zipper kinase (MELK) to be significantly overexpressed in the great majority of breast cancer cells. To assess whether MELK has a role in mammary carcinogenesis, we knocked down the expression of endogenous MELK in breast cancer cell lines using mammalian vector-based RNA interference. Furthermore, we identified a long isoform of Bcl-G (Bcl-GL), a pro-apoptotic member of the Bcl-2 family, as a possible substrate for MELK by pull-down assay with recombinant wild-type and kinase-dead MELK. Finally, we performed TUNEL assays and FACS analysis, measuring proportions of apoptotic cells, to investigate whether MELK is involved in the apoptosis cascade through the Bcl-GL-related pathway. Northern blot analyses on multiple human tissues and cancer cell lines demonstrated that MELK was overexpressed at a significantly high level in a great majority of breast cancers and cell lines, but was not expressed in normal vital organs (heart, liver, lung and kidney). Suppression of MELK expression by small interfering RNA significantly inhibited growth of human breast cancer cells. We also found that MELK physically interacted with Bcl-GL through its amino-terminal region. Immunocomplex kinase assay showed that Bcl-GL was specifically phosphorylated by MELK in vitro. TUNEL assays and FACS analysis revealed that overexpression of wild-type MELK suppressed Bcl-GL-induced apoptosis, while that of D150A-MELK did not. Our findings suggest that the kinase activity of MELK is likely to affect mammary carcinogenesis through inhibition of the pro-apoptotic function of Bcl-GL. The kinase activity of MELK could be a promising molecular target for development of therapy for patients with breast cancers.
E Hirota, L Yan, T Tsunoda, S Ashida, M Fujime, T Shuin, T Miki, Y Nakamura and Toyomasa Katagiri : Genome-wide gene expression profiles of clear cell renal cell carcinoma; Identification of molecular targets for treatment of renal cell carcinoma, International Journal of Oncology, Vol.29, No.4, 799-827, 2006.
(要約)
In order to clarify the molecular mechanism involved in renal carcinogenesis, and to identify molecular targets for diagnosis and treatment, we analyzed genome-wide gene expression profiles of 15 surgical specimens of clear cell renal cell carcinoma (RCC), compared to normal renal cortex, using a combination of laser microbeam microdissection (LMM) with a cDNA microarray representing 27,648 genes. We identified 257 genes that were commonly up-regulated and 721 genes that were down-regulated in RCCs. None of top 24 up-regulated genes that showed most significant differences in informative RCC-cases were included in previous reports describing expression profiles of RCC using RNAs isolated from bulk tissues. These findings suggest that it is important to purify as much as possible the populations of cancerous and normal epithelial cells obtained from surgical specimens. Among the significantly-transactivated genes, we focused on Semaphorin 5B (SEMA5B) and knocked-down its expression in RCC cells by small-interfering RNA (siRNA). Effective down-regulation of its expression levels in RCC cells significantly attenuated RCC cell viability. In conclusion, our data should be helpful for a better understanding of the tumorigenesis of RCC and should contribute to the development of diagnostic tumor markers and molecular-targeting therapy for patients with RCC.
JH Park, ML Lin, T Nishidate, Y Nakamura and Toyomasa Katagiri : PDZ-binding kinase/T-LAK cell-originated protein kinase, a putative cancer/testis antigen having an oncogenic activity in breast cancer, Cancer Research, Vol.66, No.18, 9186-95, 2006.
(要約)
Breast cancer is one of the most common cancers among women. To discover molecular targets that are applicable for development of novel breast cancer therapy, we previously did genome-wide expression profile analysis of 81 breast cancers and found dozens of genes that were highly and commonly up-regulated in breast cancer cells. Among them, we here focused on one gene that encodes PDZ-binding kinase/T-LAK cell-originated protein kinase (PBK/TOPK), including a kinase domain. Northern blot analyses using mRNAs of normal human organs, breast cancer tissues, and cancer cell lines indicated this molecule to be a novel cancer/testis antigen. Reduction of PBK/TOPK expression by small interfering RNA resulted in significant suppression of cell growth probably due to dysfunction in the cytokinetic process. Immunocytochemical analysis with anti-PBK/TOPK antibody implicated a critical role of PBK/TOPK in an early step of mitosis. PBK/TOPK could phosphorylate histone H3 at Ser10 in vitro and in vivo, and mediated its growth-promoting effect through histone H3 modification. Because PBK/TOPK is the cancer/testis antigen and its kinase function is likely to be related to its oncogenic activity, we suggest PBK/TOPK to be a promising molecular target for breast cancer therapy.
M Iiizumi, M Hosokawa, A Takehara, S Chung, T Nakamura, Toyomasa Katagiri, H Eguchi, H Ohigashi, O Ishikawa, Y Nakamura and H Nakagawa : EphA4 receptor, overexpressed in pancreatic ductal adenocarcinoma, promotes cancer cell growth, Cancer Science, Vol.97, No.11, 1211-1216, 2006.
(要約)
To isolate novel diagnostic markers and drug targets for pancreatic ductal adenocarcinoma (PDAC), we previously performed expression profile analysis of PDAC cells using a genome-wide cDNA microarray combined with laser microdissection. Among dozens of up-regulated genes identified in PDAC cells, we herein focused on one tyrosine kinase receptor, Eph receptor A4 (EphA4), as a molecular target for PDAC therapy. Immunohistochemical analysis validated EphA4 overexpression in approximately half of the PDAC tissues. To investigate its biological function in PDAC cells, we knocked down EphA4 expression by siRNA, which drastically attenuated PDAC cell viability. In concordance with the siRNA experiment, PDAC-derivative cells that were designed to constitutively express exogenous EphA4 showed a more rapid growth rate than cells transfected with mock vector, suggesting a growth-promoting effect of EphA4 on PDAC cells. Furthermore, the expression analysis for ephrin ligand family members indicated the coexistence of ephrinA3 ligand in PDAC cells with EphA4 receptor, and knockdown of ephrinA3 by siRNA also attenuated PDAC cell viability. These results suggest that the EphA4-ephrinA3 pathway is likely to be a promising molecular target for pancreatic cancer therapy.
A Takehara, H Eguchi, H Ohigashi, O Ishikawa, T Kasugai, M Hosokawa, Toyomasa Katagiri, Y Nakamura and H Nakagawa : Novel tumor marker REG4 detected in serum of patients with resectable pancreatic cancer and feasibility for antibody therapy targeting REG4, Cancer Science, Vol.97, No.11, 1191-1197, 2006.
(要約)
Pancreatic ductal adenocarcinoma (PDAC) shows the worst mortality rate among common malignancies, with a 5-year survival rate of only 4%, and the majority of PDAC patients are diagnosed at an advanced stage in which no effective therapy is available at present. Although the proportion of curable cases is still not so high, surgical resection of early stage PDAC is the only way to cure the disease. Hence, establishment of a screening strategy to detect early stage PDAC by novel serological markers is required urgently, and development of novel molecular therapies for PDAC treatment is also eagerly expected. We here report overexpression of REG4, a new member of the regenerating islet-derived (REG) family, in PDAC cells on the basis of genome-wide cDNA microarray analysis as well as reverse transcription-polymerase chain reaction and immunohistochemical analysis. We also detected significant elevation of REG4 in the serum of some patients with early-stage PDAC using our enzyme-linked immunosorbent assay system, indicating the possibility of REG4 as a new serological marker of PDAC. Furthermore, we found that knockdown of endogenous REG4 expression in PDAC cell lines with small interfering RNA caused a decrease in cell viability. Concordantly, addition of recombinant REG4 to the culture medium enhanced growth of a PDAC cell line in a dose-dependent manner. A monoclonal antibody against REG4 neutralized its growth-promoting effects and attenuated significantly the growth of PDAC cells. These findings indicate that REG4 is a promising tumor marker to screen early-stage PDAC, and also that neutralization of REG4 by the antibody may offer novel potential tools for the treatment of PDAC.
S Ashida, M Furihata, Toyomasa Katagiri, K Tamura, Y Anazawa, M Yoshioka, T Miki, T Fujioka, T Shuin, Y Nakamura and H Nakagawa : Expression of novel molecules, MICAL2-PV (MICAL2 prostate cancer variants), increases with high Gleason score and prostate cancer progression, Clinical Cancer Research, Vol.12, No.9, 2767-2773, 2006.
(要約)
The aim of this study is to identify novel molecular targets for development of novel treatment or diagnostic markers of prostate cancer through genome-wide cDNA microarray analysis of prostate cancer cells purified by laser microdissection. Here, we identified molecule interacting with CasL-2 prostate cancer variants (MICAL2-PV), novel splicing variants of MICAL2, showing overexpression in prostate cancer cells. Immunohistochemical analysis using an antibody generated specific to MICAL2-PV revealed that MICAL2-PV was expressed in the cytoplasm of cancer cells with various staining patterns and intensities, whereas it was not or hardly detectable in adjacent normal prostate epithelium or prostatic intraepithelial neoplasia. Interestingly, immunohistochemical analysis of 105 prostate cancer specimens on the tissue microarray indicated that MICAL2-PV expression status was strongly correlated with Gleason scores (P < 0.0001) or tumor classification (P < 0.0001). Furthermore, the expression levels of MICAL2-PVs were also concordant to those of c-Met, a marker of tumor progression, with statistical significance (P = 0.0018). To investigate its potential of molecular therapeutic target for prostate cancers, we knocked down endogenous MICAL2-PVs in prostate cancer cells by small interfering RNA, which resulted in the significant reduction of prostate cancer cell viability. Our findings suggest that MICAL2-PV is likely to be involved in cancer progression of prostate cancer and could be a candidate as a novel molecular marker and/or target for treatment of prostate cancers with high Gleason score.
(キーワード)
Adaptor Proteins, Signal Transducing / Alternative Splicing / Base Sequence / Cell Line, Tumor / Cytoskeletal Proteins / DNA Primers / Disease Progression / Genetic Variation / Humans / Immunohistochemistry / Intracellular Signaling Peptides and Proteins / LIM Domain Proteins / Male / Prostatic Neoplasms
T Kikuchi, Y Daigo, N Ishikawa, Toyomasa Katagiri, T Tsunoda, S Yoshida and Y Nakamura : Expression profiles of metastatic brain tumor from lung adenocarcinomas on cDNA microarray, International Journal of Oncology, Vol.28, No.4, 799-805, 2006.
(要約)
Distant metastasis is one of the crucial parameters determining the type of treatment and prognosis of patients. Previous studies discovered important factors involved in multiple steps of metastasis, the precise mechanisms of metastasis still remain to be clarified. To identify genes associated with this complicated biological feature of cancer, we analyzed expression profiles of 16 metastatic brain tumors derived from primary lung adenocarcinoma (ADC) using cDNA microarray representing 23,040 genes. We applied bioinformatic algorithm to compare the expression data of these 16 brain metastatic loci with those of 37 primary NSCLCs including 22 ADCs, and found that metastatic tumor cells has very different characteristics of gene expression patterns from primary ones. Two hundred and forty-four genes that showed significantly different expression levels between the two groups included plasma membrane bounding proteins, cellular antigens, and cytoskeletal proteins that might play important roles in altering cell-cell communication, attachment, and cell motility, and enhance the metastatic ability of cancer cells. Our results provide valuable information for development of predictive markers as well as novel therapeutic target molecules for metastatic brain tumor of ADC of the lung.
R Hamamoto, FP Silva, M Tsuge, T Nishidate, Toyomasa Katagiri, Y Nakamura and Y Furukawa : Enhanced SMYD3 expression is essential for the growth of breast cancer cells, Cancer Science, Vol.97, No.2, 113-118, 2006.
(要約)
We previously reported that upregulation of SMYD3, a histone H3 lysine-4-specific methyltransferase, plays a key role in the proliferation of colorectal carcinoma (CRC) and hepatocellular carcinoma (HCC). In the present study, we reveal that SMYD3 expression is also elevated in the great majority of breast cancer tissues. Similarly to CRC and HCC, silencing of SMYD3 by small interfering RNA to this gene resulted in the inhibited growth of breast cancer cells, suggesting that increased SMYD3 expression is also essential for the proliferation of breast cancer cells. Moreover, we show here that SMYD3 could promote breast carcinogenesis by directly regulating expression of the proto-oncogene WNT10B. These data imply that augmented SMYD3 expression plays a crucial role in breast carcinogenesis, and that inhibition of SMYD3 should be a novel therapeutic strategy for treatment of breast cancer.
JR Morris, L Pangon, C Boutell, Toyomasa Katagiri, NH Keep and E Solomon : Genetic analysis of BRCA1 ubiquitin ligase activity and its relationship to breast cancer susceptibility, Human Molecular Genetics, Vol.15, No.4, 599-606, 2006.
(要約)
The N-terminus of the Breast Cancer-1 predisposition protein (BRCA1) associates with the BRCA1-associated RING domain-1 protein (BARD1) to form a heterodimer, which exhibits ubiquitin ligase activity that is abrogated by known cancer-associated BRCA1 missense mutations. The majority of missense substitutions identified in patients with a personal or a family history of disease have not been followed in pedigrees, nor there is a functional understanding of their impact. We have examined, by extensive missense substitution, the interaction of BRCA1 with components that contribute to its ubiquitin ligase activity, BARD1 and the E2 ubiquitin-conjugating enzyme, UbcH5a. Selection from a randomly generated library of BRCA1 missense mutations for variants that inhibit the interaction with these components identified substitutions in residues found altered in patient DNA, indicating a correlation between loss of component-binding and propensity to disease development. We further show that the BRCA1:E2 interaction is sensitive to substitutions in all structural elements of the BRCA1 N-terminus, whereas the BARD1 interaction is sensitive to a subset of BRCA1 substitutions, which also inhibit E2-binding. Patient variants that inhibit the BRCA1:E2 interaction show loss of ubiquitin ligase activity and correlate with disease susceptibility and theoretical predictions of pathogenicity. These data link the loss of ubiquitin ligase activity, through loss of E2-binding, to the majority of non-polymorphic patient variants described within the N-terminus of BRCA1 and illustrate the likely significant role of BRCA1 ubiquitin ligase activity in tumour suppression.
(キーワード)
BRCA1 Protein / Dimerization / Female / Genetic Predisposition to Disease / Humans / Iron-Binding Proteins / Male / Multiprotein Complexes / Mutation, Missense / Neoplasms / Protein Binding / Protein Structure, Tertiary / Tumor Suppressor Proteins / Ubiquitin-Conjugating Enzymes / Ubiquitin-Protein Ligases
Seiji Tachiiri, Toyomasa Katagiri, Tatsuhiko Tsunoda, Natsuo Oya, Masahiro Hiraoka and Yusuke Nakamura : Analysis of gene-expression profiles after gamma irradiation of normal human fibroblasts, International Journal of Radiation Oncology*Biology*Physics, Vol.64, No.1, 272-279, 2006.
(要約)
To understand comprehensive transcriptional profile of normal human fibroblast in response to irradiation. To identify genes whose expression is influenced by gamma radiation, we used a cDNA microarray to analyze expression of 23,000 genes in normal human fibroblasts at 7 timepoints (1, 3, 6, 12, 24, 48, and 72 hours) after 5 different doses (0.5, 2, 5, 15, and 50 Gy) of exposure. Among the genes that showed altered expression patterns, some were already known to be regulated by irradiation, for instance ODC, EGR1, FGF2, PCNA, PKC, and several p53-target genes, including p53DINP1, BTG2, GADD45, and MDM2. The time course of each dose showed that from 350 to 600 genes were affected as to their expression; induction profiles characteristic to each dose were demonstrated. Of the total identified, only 89 genes were up-regulated; the vast majority was down-regulated over the 72-hour time course. We identified 21 genes that were distinctly induced by irradiation; 11 of them were functionally known, and 6 of those were p53-target genes. The results underscored the complexity of the transcriptional responses to irradiation, and the data should serve as a basis for global characterization of radiation-regulated genes and pathways.
S Nagayama, C Fukukawa, Toyomasa Katagiri, T Okamoto, T Aoyama, N Oyaizu, M Imamura, J Toguchida and Y Nakamura : Therapeutic potential of antibodies against FZD10, a cell-surface protein, for synovial sarcomas, Oncogene, Vol.24, No.41, 6201-6212, 2005.
(要約)
Genome-wide expression profiling revealed overexpression of the gene encoding frizzled homologue 10 (FZD 10), a cell-surface receptor for molecules in the Wnt pathway, as a potential contributor to synovial sarcomas (SS). Northern blotting and immunohistochemical staining confirmed that expression levels of FZD 10 were very high in nearly all SS tumors and cell lines examined but absent in most normal organs or in some cancers arising in other tissues. Treatment of human SS cells with small-interfering RNA (siRNA) to FZD 10 decreased the amount of its product and suppressed growth of SS cells. Moreover, a polyclonal antibody specifically recognizing the extracellular domain (ECD) of FZD 10 was markedly effective in mediating ADCC against FZD 10-overexpressing synovial sarcoma cells in vitro. Injection of the antibody into SS xenografts in nude mice attenuated tumor growth, and TUNEL assays revealed clusters of apoptotic cells in antibody-treated xenografts. Taken together, these findings suggest that a humanized antibody against FZD 10 might be a promising treatment for patients with tumors that overexpress FZD 10; minimal or no adverse reactions would be expected because FZD 10 protein is not abundant in vital organs.
A Togashi, Toyomasa Katagiri, S Ashida, T Fujioka, O Maruyama, Y Wakumoto, Y Sakamoto, M Fujime, Y Kawachi, T Shuin and Y Nakamura : Hypoxia-inducible protein 2 (HIG2), a novel diagnostic marker for renal cell carcinoma (RCC) and potential target for molecular therapy, Cancer Research, Vol.65, No.11, 4817-4826, 2005.
(要約)
To identify molecules to serve as diagnostic markers for renal cell carcinoma (RCC) and as targets for novel therapeutic drugs, we investigated genome-wide expression profiles of RCCs using a cDNA microarray. We subsequently confirmed that hypoxia-inducible protein-2 (HIG2) was expressed exclusively in RCCs and fetal kidney. Induction of HIG2 cDNA into COS7 cells led to secretion of the gene product into culture medium and resulted in enhancement of cell growth. Small interfering RNA effectively inhibited expression of HIG2 in human RCC cells that endogenously expressed high levels of the protein and significantly suppressed cell growth. Moreover, addition of polyclonal anti-HIG2 antibody into culture medium induced apoptosis in RCC-derived cell lines. By binding to an extracellular domain of frizzled homologue 10 (FZD10), HIG2 protein enhanced oncogenic Wnt signaling and its own transcription, suggesting that this product is likely to function as an autocrine growth factor. ELISA analysis of clinical samples identified secretion of HIG2 protein into the plasma of RCC patients even at an early stage of tumor development, whereas it was detected at significantly lower levels in healthy volunteers or patients with chronic glomerulonephritis. The combined evidence suggests that this molecule represents a promising candidate for development of molecular-targeting therapy and could serve as a prominent diagnostic tumor marker for patients with renal carcinomas.
Y Anazawa, H Nakagawa, M Furihara, S Ashida, K Tamura, H Yoshioka, T Shuin, T Fujioka, Toyomasa Katagiri and Y Nakamura : PCOTH, a novel gene overexpressed in prostate cancers, promotes prostate cancer cell growth through phosphorylation of oncoprotein TAF-Ibeta/SET., Cancer Research, Vol.65, No.11, 4578-4586, 2005.
(要約)
Through genome-wide cDNA microarray analysis coupled with microdissection of prostate cancer cells, we identified a novel gene, prostate collagen triple helix (PCOTH), showing overexpression in prostate cancer cells and its precursor cells, prostatic intraepithelial neoplasia (PIN). Immunohistochemical analysis using polyclonal anti-PCOTH antibody confirmed elevated expression of PCOTH, a 100-amino-acid protein containing collagen triple-helix repeats, in prostate cancer cells and PINs. Knocking down PCOTH expression by small interfering RNA (siRNA) resulted in drastic attenuation of prostate cancer cell growth, and concordantly, LNCaP derivative cells that were designed to constitutively express exogenous PCOTH showed higher growth rate than LNCaP cells transfected with mock vector, suggesting the growth-promoting effect of PCOTH on prostate cancer cell. To investigate the biological mechanisms of this growth-promoting effect, we applied two-dimensional differential gel electrophoresis (2D-DIGE) to analyze the phospho-protein fractions in LNCaP cells transfected with PCOTH. We found that the phosphorylation level of oncoprotein TAF-Ibeta/SET was significantly elevated in LNCaP cells transfected with PCOTH than control LNCaP cells, and these findings were confirmed by Western blotting and in-gel kinase assay. Furthermore, knockdown of endogenous TAF-Ibeta expression by siRNA also attenuated viability of prostate cancer cells as well. These findings suggest that PCOTH is involved in growth and survival of prostate cancer cells thorough, in parts, the TAF-Ibeta pathway, and that this molecule should be a promising target for development of new therapeutic strategies for prostate cancers.
K Obama, K Ura, M Li, Toyomasa Katagiri, T Tsunoda, A Nomura, S Satoh, Y Nakamura and Y Furukawa : Genome-wide analysis of gene expression in human intrahepatic cholangiocarcinoma, Hepatology, Vol.41, No.6, 1339-48, 2005.
(要約)
Intrahepatic cholangiocarcinoma is a neoplasm arising in the liver, and its incidence is increasing in Japan as well as in Western countries. Prognosis of patients with this type of tumor remains unsatisfactory because no effective chemotherapeutic drugs are available, we have no sensitive tumor markers to detect this tumor in its early stage, and it is difficult to identify a high-risk group for the disease. To clarify the molecular mechanism of tumorigenesis and identify molecular targets for diagnosis and treatment, we analyzed global gene-expression profiles of 25 intrahepatic cholangiocarcinomas using tumor cell populations purified by laser microbeam microdissection and a cDNA microarray containing 27,648 genes. We identified 52 genes that were commonly upregulated and 421 that were downregulated in intrahepatic cholangiocarcinomas compared with noncancerous biliary epithelial cells. From the 52 upregulated genes, we selected P-cadherin and survivin for further investigation and corroborated enhanced expression of their products in cancer tissues by immunohistochemical staining. Furthermore, comparison between tumors with lymph node metastasis and those without metastasis identified 30 genes that were associated with lymph node involvement. In conclusion, these data should be helpful for a better understanding of the tumorigenesis of intrahepatic cholangiocarcinoma and should contribute to the development of diagnostic and therapeutic strategies for this type of tumor. Supplementary material for this article can be found on the HEPATOLOGY website (http://www.interscience.wiley.com/jpages/0270-9139/suppmat/index.html).
K Taniuchi, H Nakagawa, M Hosokawa, T Nakamura, H Eguchi, H Ohigashi, O Ishikawa, Toyomasa Katagiri and Y Nakamura : Overexpressed P-cadherin/CDH3 promotes motility of pancreatic cancer cells by interacting with p120ctn and activating rho-family GTPases, Cancer Research, Vol.65, No.8, 3092-3099, 2005.
(要約)
P-Cadherin/CDH3 belongs to the family of classic cadherins that are engaged in various cellular activities including motility, invasion, and signaling of tumor cells, in addition to cell adhesion. However, the biological roles of P-cadherin itself are not fully characterized. Based on information derived from a previous genome-wide cDNA microarray analysis of microdissected pancreatic ductal adenocarcinoma (PDAC), we focused on P-cadherin as one of the genes most strongly overexpressed in the great majority of PDACs. To investigate the consequences of overexpression of P-cadherin in terms of pancreatic carcinogenesis and tumor progression, we used a P-cadherin-deficient PDAC cell line, Panc-1, to construct a cell line (Panc1-CDH3) that stably overexpressed P-cadherin. Induction of P-cadherin in Panc1-CDH3 increased the motility of the cancer cells, but a blocking antibody against P-cadherin suppressed the motility in vitro. Overexpression of P-cadherin was strongly associated with cytoplasmic accumulation of one of the catenins, p120ctn, and cadherin switching in PDAC cells. Moreover, P-cadherin-dependent activation of cell motility was associated with activation of Rho GTPases, Rac1 and Cdc42, through accumulation of p120ctn in cytoplasm and cadherin switching. These findings suggest that overexpression of P-cadherin is likely to be related to the biological aggressiveness of PDACs; blocking of P-cadherin activity or its associated signaling could be a novel therapeutic approach for treatment of aggressive pancreatic cancers.
T Ishibe, T Nakayama, T Okamoto, T Aoyama, K Nishijo, KR Shibata, Y Shima, S Nagayama, Toyomasa Katagiri, Y Nakamura, T Nakamura and J. Toguchida : Disruption of fibroblast growth factor signal pathway inhibits the growth of synovial sarcomas: potential application of signal inhibitors to molecular target therapy, Clinical Cancer Research, Vol.11, No.7, 2702-12, 2005.
(要約)
Synovial sarcoma is a soft tissue sarcoma, the growth regulatory mechanisms of which are unknown. We investigated the involvement of fibroblast growth factor (FGF) signals in synovial sarcoma and evaluated the therapeutic effect of inhibiting the FGF signal. The expression of 22 FGF and 4 FGF receptor (FGFR) genes in 18 primary tumors and five cell lines of synovial sarcoma were analyzed by reverse transcription-PCR. Effects of recombinant FGF2, FGF8, and FGF18 for the activation of mitogen-activated protein kinase (MAPK) and the growth of synovial sarcoma cell lines were analyzed. Growth inhibitory effects of FGFR inhibitors on synovial sarcoma cell lines were investigated in vitro and in vivo. Synovial sarcoma cell lines expressed multiple FGF genes especially those expressed in neural tissues, among which FGF8 showed growth stimulatory effects in all synovial sarcoma cell lines. FGF signals in synovial sarcoma induced the phosphorylation of extracellular signal-regulated kinase (ERK1/2) and p38MAPK but not c-Jun NH2-terminal kinase. Disruption of the FGF signaling pathway in synovial sarcoma by specific inhibitors of FGFR caused cell cycle arrest leading to significant growth inhibition both in vitro and in vivo. Growth inhibition by the FGFR inhibitor was associated with a down-regulation of phosphorylated ERK1/2 but not p38MAPK, and an ERK kinase inhibitor also showed growth inhibitory effects for synovial sarcoma, indicating that the growth stimulatory effect of FGF was transmitted through the ERK1/2. FGF signals have an important role in the growth of synovial sarcoma, and inhibitory molecules will be of potential use for molecular target therapy in synovial sarcoma.
R Takata, Toyomasa Katagiri, M Kanehira, T Tsunoda, T Shuin, T Miki, M Namiki, K Kohri, Y Matsushita, T Fujioka and Y Nakamura : Predicting response to methotrexate, vinblastine, doxorubicin, and cisplatin neoadjuvant chemotherapy for bladder cancers through genome-wide gene expression profiling, Clinical Cancer Research, Vol.11, No.7, 2625-36, 2005.
(要約)
Neoadjuvant chemotherapy for invasive bladder cancer, involving a regimen of methotrexate, vinblastine, doxorubicin, and cisplatin (M-VAC), can improve the resectability of larger neoplasms for some patients and offer a better prognosis. However, some suffer severe adverse drug reactions without any effect, and no method yet exists for predicting the response of an individual patient to chemotherapy. Our purpose in this study is to establish a method for predicting response to the M-VAC therapy. We analyzed gene expression profiles of biopsy materials from 27 invasive bladder cancers using a cDNA microarray consisting of 27,648 genes, after populations of cancer cells had been purified by laser microbeam microdissection. We identified dozens of genes that were expressed differently between nine "responder" and nine "nonresponder" tumors; from that list we selected the 14 "predictive" genes that showed the most significant differences and devised a numerical prediction scoring system that clearly separated the responder group from the nonresponder group. This system accurately predicted the drug responses of 8 of 9 test cases that were reserved from the original 27 cases. Because real-time reverse transcription-PCR data were highly concordant with the cDNA microarray data for those 14 genes, we developed a quantitative reverse transcription-PCR-based prediction system that could be feasible for routine clinical use. Our results suggest that the sensitivity of an invasive bladder cancer to the M-VAC neoadjuvant chemotherapy can be predicted by expression patterns in this set of genes, a step toward achievement of "personalized therapy" for treatment of this disease.
K Taniuchi, H Nakagawa, T Nakamura, H Eguchi, H Ohigashi, O Ishikawa, Toyomasa Katagiri and Y Nakamura : Down-regulation of RAB6KIFL/KIF20A, a kinesin involved with membrane trafficking of discs large homologue 5, can attenuate growth of pancreatic cancer cell, Cancer Research, Vol.65, No.1, 105-112, 2005.
(要約)
To identify novel molecular targets for treatment of pancreatic ductal adenocarcinoma (PDAC), we generated precise gene expression profiles of PDACs on a genome-wide cDNA microarray after populations of tumor cells were purified by laser microdissection. Through functional analysis of genes that were transactivated in PDACs, we identified RAB6KIFL as a candidate for development of drugs to treat PDACs at the molecular level. Knockdown of endogenous RAB6KIFL expression in PDAC cell lines by small interfering RNA drastically attenuated growth of those cells, suggesting an essential role for the gene product in maintaining viability of PDAC cells. RAB6KIFL belongs to the kinesin superfamily of motor proteins, which have critical functions in trafficking of molecules and organelles. Proteomics analyses using a polyclonal anti-RAB6KIFL antibody identified one of the cargoes transported by RAB6KIFL as discs, large homologue 5 (DLG5), a scaffolding protein that may link the vinexin-beta-catenin complex at sites of cell-cell contact. Like RAB6KIFL, DLG5 was overexpressed in PDACs, and knockdown of endogenous DLG5 by small interfering RNA significantly suppressed the growth of PDAC cells as well. Decreased levels of endogenous RAB6KIFL in PDAC cells altered the subcellular localization of DLG5 from cytoplasmic membranes to cytoplasm. Our results imply that collaboration of RAB6KIFL and DLG5 is likely to be involved in pancreatic carcinogenesis. These molecules should be promising targets for development of new therapeutic strategies for PDACs.
(キーワード)
Cell Division / Cell Line / Cell Line, Tumor / Gene Expression Regulation, Neoplastic / Humans / Kidney / Kinesin / Kinetics / Mass Spectrometry / Membrane Proteins / Pancreatic Neoplasms / Protein Transport / RNA, Small Interfering / Reverse Transcriptase Polymerase Chain Reaction / Tubulin / Tumor Suppressor Proteins
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 15665285
K Okada, E Hirota, Y Mizutani, T Fujioka, T Shuin, T Miki, Y Nakamura and Toyomasa Katagiri : Oncogenic role of NALP7 in testicular seminomas, Cancer Science, Vol.95, No.12, 949-954, 2004.
(要約)
To isolate novel molecular targets for treatment of testicular germ cell tumor (TGCT), we performed genome-wide expression profile analysis of testicular seminomas using a cDNA microarray. We here report identification of NACHT, leucine-rich repeat and PYD containing 7 (NALP7 ), that was significantly transactivated in testicular seminomas. Subsequent semi-quantitative RT-PCR and northern blot analyses confirmed an approximately 3.3-kb transcript that was expressed exclusively in testis, although the expression level of this gene in normal testis was much lower than in tumor cells, suggesting an important role of this gene in germ-cell proliferation. Immunohistochemical analysis using anti-NALP7 polyclonal antibody detected the endogenous NALP7 protein in the cytoplasm of embryonal carcinoma cells and testicular seminoma tissues. Transfection of small interfering RNA (siRNA) for NALP7 significantly reduced the NALP7 expression and resulted in growth suppression of testicular germ-cell tumors. These findings imply that NALP7 may play a crucial role in cell proliferation, as well as testicular tumorigenesis, and it appears to be a promising candidate for development of targeted therapy for TGCTs.
H Kamiyama, K Kurosaki, M Kurimoto, Toyomasa Katagiri, Y Nakamura, M Kurokawa, H Sato, S Endo and K Shiraki : Herpes simplex virus-induced, death receptor-dependent apoptosis and regression of transplanted human cancers, Cancer Science, Vol.95, No.12, 990-998, 2004.
(要約)
Inoculation of a live attenuated herpes simplex virus (HSV) vector, betaH1, into human U87MG glioblastoma cells transplanted into athymic nude mice induced complete regression of tumors. The infected cells underwent histochemically confirmed apoptosis without lymphocyte infiltration after expressing CD30, CD30 ligand (CD30L), tumor necrosis factor (TNF)-alpha, TNF receptor 1 (TNF-R1), FAS, and FAS ligand (FAS-L) with activation of caspases 3 and 8. Induction of the transcripts of these receptors and ligands in inoculated tumors was confirmed by quantitative RT-PCR. To examine the specificity of apoptosis in the transplanted tumor, we inoculated betaH1 into transplanted human lung, breast, gastric, and colon cancer tumors, and similar tumor regression with apoptosis was observed in all tumors. We analyzed the roles of expression of CD30, CD30L, TNF-alpha, TNF-R1, FAS, and FAS-L in the tumors, and found that HSV-induced apoptosis was suppressed by the respective antibodies. These findings indicate that the CD30/CD30L, TNF-alpha/TNF-R1, and FAS/FAS-L interactions resulted in apoptosis and tumor regression in immunocompromised mice. In addition to the death receptor-dependent apoptosis induced by HSV, the expressed ligands and receptors might enhance the susceptibility of tumor cells to cell-mediated cyto-toxicity and augment the activation of tumor-killing lymphocytes in immunocompetent models.
Tetsuya Nakatsura, Hiroyuki Komori, Tatsuko Kubo, Yoshihiro Yoshitake, Satoru Senju, Toyomasa Katagiri, Yoichi Furukawa, Michio Ogawa, Yusuke Nakamura and Yasuharu Nishimura : Mouse homologue of a novel human oncofetal antigen, glypican-3, evokes T-cell-mediated tumor rejection without autoimmune reactions in mice, Clinical Cancer Research, Vol.10, No.24, 8630-40, 2004.
(要約)
We recently identified glypican-3 (GPC3) overexpressed specifically in human hepatocellular carcinoma, as based on cDNA microarray analysis of 23,040 genes, and we reported that GPC3 is a novel tumor marker for human hepatocellular carcinoma and melanoma. GPC3, expressed in almost all hepatocellular carcinomas and melanomas, but not in normal tissues except for placenta or fetal liver, is a candidate of ideal tumor antigen for immunotherapy. In this study, we attempted to identify a mouse GPC3 epitope for CTLs in BALB/c mice, and for this, we set up a preclinical study to investigate the usefulness of GPC3 as a target for cancer immunotherapy in vivo. We identified a mouse GPC3-derived and Kd- restricted CTL epitope peptide in BALB/c mice. Inoculation of this GPC3 peptide-specific CTL into s.c. Colon26 cancer cells transfected with mouse GPC3 gene (C26/GPC3) led to rejection of the tumor in vivo, and i.v. inoculation of these CTLs into sublethally irradiated mice markedly inhibited growth of an established s.c. tumor. Inoculation of bone marrow-derived dendritic cells pulsed with this peptide prevented the growth of s.c. and splenic C26/GPC3 accompanied with massive infiltration of CD8+ T cells into tumors. Evidence of autoimmune reactions was never observed in surviving mice that had rejected tumor cell challenges. We found the novel oncofetal protein GPC3 to be highly immunogenic in mice and elicited effective antitumor immunity with no evidence of autoimmunity. GPC3 is useful not only for diagnosis of hepatocellular carcinoma and melanoma but also for possible immunotherapy or prevention of these tumors.
Y Yoshitake, T Nakatsura, M Monji, S Senju, H Matsuyoshi, H Tsukamoto, S Hosaka, H Komori, D Fukuma, Y Ikuta, Toyomasa Katagiri, Y Furukawa, H Ito, M Shinohara, Y Nakamura and Y Nishimura : Proliferation potential-related protein, an ideal esophageal cancer antigen for immunotherapy, identified using complementary DNA microarray analysis, Clinical Cancer Research, Vol.10, No.19, 6437-48, 2004.
(要約)
To establish effective antitumor immunotherapy for esophageal cancer, we tried to identify an useful target antigen of esophageal cancer. We did cDNA microarray analysis to find a novel candidate antigen, proliferation potential-related protein (PP-RP). We examined cytotoxicity against tumor cells in vitro and in vivo of CTLs specific to PP-RP established from esophageal cancer patients. In 26 esophageal cancer tissues, an average of relative ratio of the expression of the PP-RP mRNA in cancer cells versus adjacent normal esophageal tissues was 396.2. Immunohistochemical analysis revealed that, in 20 of the 22 esophageal cancer tissues, PP-RP protein was strongly expressed only in the cancer cells and not so in normal esophageal epithelial cells. PP-RP protein contains 10 epitopes recognized by HLA-A24-restricted CTLs. These CTLs, generated from HLA-A24-positive esophageal cancer patients, had cytotoxic activity against cancer cell lines positive for both PP-RP and HLA-A24. Furthermore, adoptive transfer of the PP-RP-specific CTL line inhibited the growth of a human esophageal cancer cell line engrafted in nude mice. The expression of PP-RP in esophageal cancer cells was significantly higher than in normal cells, and the CTLs recognizing PP-RP killed tumor cells in vitro and also showed tumor rejection effects in a xenograft model. Therefore, PP-RP may prove to be an ideal tumor antigen useful for diagnosis and immunotherapy for patients with esophageal cancer. cDNA microarray analysis is a useful method to identify ideal tumor-associated antigens.
T Nishidate, Toyomasa Katagiri, ML Lin, Y Mano, Y Miki, F Kasumi, M Yoshimoto, T Tsunoda, K Hirata and Y Nakamura : Genome-wide gene-expression profiles of breast-cancer cells purified with laser microbeam microdissection: identification of genes associated with progression and metastasis, International Journal of Oncology, Vol.25, No.4, 797-819, 2004.
(要約)
Breast carcinoma is a complex disease characterized by accumulation of multiple genetic alterations, and the understanding of the molecular basis of mammary tumorigenesis is still incomplete. In this study we analyzed gene-expression profiles of 81 surgical specimens of 12 ductal carcinoma in situ (DCIS) and 69 invasive ductal carcinoma (IDC). After applying laser-microbeam micro-dissection to all samples we achieved 98-99% pure populations of breast cancer cells, and of normal breast epithelial cells used as controls. A cDNA-microarray analysis of 23,040 genes in these samples and a subsequent unsupervised hierarchical clustering distinguished two tumor groups, mainly in terms of estrogen-receptor (ER) status. We then undertook a supervised analysis and identified 325 genes that were commonly either up- or down-regulated in both pathologically discrete stages (DCIS and IDC), indicating that these genes might play important roles in malignant transformation of breast ductal cells. In addition, we searched invasion-associated gene candidates whose expression was altered in IDC, but not in DCIS, and identified 24 up-regulated genes and 41 down-regulated genes. Furthermore, we identified 34 genes that were expressed differently in tumors from patients with lymph node metastasis as opposed to no metastasis. On that basis we developed a scoring system that correlated well with the metastatic status. Tumors from all of the 37 test patients with lymph-node metastasis yielded positive scores by our definition, whereas 38 of the 40 tumors (95%) without lymph node metastasis had negative scores. Our data should provide useful information for identifying predictive markers for invasion or metastasis, and suggest potential target molecules for treatment of breast cancers.
Y Harima, A Togashi, K Horikoshi, M Imamura, M Sougawa, S Sawada, T Tsunoda, Y Nakamura and Toyomasa Katagiri : Prediction of outcome of advanced cervical cancer to thermoradiotherapy according to expression profiles of 35 genes selected by cDNA microarray analysis, International Journal of Radiation Oncology*Biology*Physics, Vol.60, No.1, 237-248, 2004.
(要約)
To identify a set of genes related to thermoradiosensitivity of cervical carcinoma and to establish a predictive method. A total of 19 patients with cervical cancer (1 with Stage IIIA, 11 with Stage IIIB, 5 with Stage IVA, and 2 with Stage IVB) who underwent definitive thermoradiotherapy between May 1995 and August 2001 were included in this study. We compared the expression profiles of 8 thermoradiosensitive and 11 thermoradioresistant tumors obtained by punch biopsy before treatment using a cDNA microarray consisting of 23,040 human genes. We selected 35 genes on the basis of a clustering analysis and confirmed the validity of these genes with a cross-validation test. Some of these genes were already known to be associated with apoptosis (BIK, TEGT, SSI-3), hypoxia-inducible genes (HIF1A, CA12), and tumor cell invasion and metastasis (CTSL, CTSB, PLAU, CD44). We developed a "predictive score" system that could clearly separate the thermoradiosensitive group from the thermoradioresistant group. These results from the treatment program between May 1995 and August 2001 showed that by using gene-expression profiles we can predict the outcome of thermoradiotherapy for advanced cervical carcinoma. A "predictive score" system was developed that could clearly separate the thermoradiosensitive group from the thermoradioresistant group. These results may eventually lead to the achievement of "personalized therapy" for this disease.
Shingo Ashida, Hidewaki Nakagawa, Toyomasa Katagiri, Mutsuo Furihata, Megumi Iiizumi, Yoshio Anazawa, Tatsuhiko Tsunoda, Ryo Takata, Kotaro Kasahara, Tsuneharu Miki, Tomoaki Fujioka, Taro Shuin and Yusuke Nakamura : Molecular features of the transition from prostatic intraepithelial neoplasia (PIN) to prostate cancer: genome-wide gene-expression profiles of prostate cancers and PINs, Cancer Research, Vol.64, No.17, 5963-5972, 2004.
(要約)
To characterize the molecular feature in prostate carcinogenesis and the putative transition from prostatic intraepithelial neoplasia (PIN) to invasive prostate cancer (PC), we analyzed gene-expression profiles of 20 PCs and 10 high-grade PINs with a cDNA microarray representing 23,040 genes. Considering the histological heterogeneity of PCs and the minimal nature of PIN lesions, we applied laser microbeam microdissection to purify populations of PC and PIN cells, and then compared their expression profiles with those of corresponding normal prostatic epithelium also purified by laser microbeam microdissection. A hierarchical clustering analysis separated the PC group from the PIN group, except for three tumors that were morphologically defined as one very-high-grade PIN and two low-grade PCs, suggesting that PINs and PCs share some molecular features and supporting the hypothesis of PIN-to-PC transition. On the basis of this hypothesis, we identified 21 up-regulated genes and 63 down-regulated genes commonly in PINs and PCs compared with normal epithelium, which were considered to be involved in the presumably early stage of prostatic carcinogenesis. They included AMACR, OR51E2, RODH, and SMS. Furthermore, we identified 41 up-regulated genes and 98 down-regulated genes in the transition from PINs to PCs; those altered genes, such as POV1, CDKN2C, EPHA4, APOD, FASN, ITGB2, LAMB2, PLAU, and TIMP1, included elements that are likely to be involved in cell adhesion or the motility of invasive PC cells. The down-regulation of EPHA4 by small interfering RNA in PC cells lead to attenuation of PC cell viability. These data provide clues to the molecular mechanisms underlying prostatic carcinogenesis, and suggest candidate genes the products of which might serve as molecular targets for the prevention and treatment of PC.
S Nagayama, M Iiizumi, Toyomasa Katagiri, J Toguchida and Y Nakamura : Identification of PDZK4, a novel human gene with PDZ domains, that is up-regulated in synovial sarcomas, Oncogene, Vol.23, No.32, 5551-7, 2004.
(要約)
In an earlier study designed to investigate molecular mechanisms of carcinogenesis in synovial sarcomas (SSs), we applied a cDNA microarray to detect human genes with significantly increased expression in SS cells. Among the genes selected in this way, we identified a novel transcript, subsequently designated PDZK4 (PDZ domain-containing 4), that was specifically upregulated in all of the 13 SS cases we examined. On Northern blots of normal human tissues, the PDZK4 transcript was expressed only in fetal brain. Immunocytochemical staining of transfected COS7 cells showed that the PDZK4 gene product localized mainly under the plasma membrane. Treatment of human SS cells with small interfering RNA (siRNA) inhibited the expression of PDZK4 and resulted in the suppression of tumor-cell growth. Induction of exogenous PDZK4 expression promoted growth of T98G and COS7 cells in which no endogenous expression of PDZK4 was observed. Taken together, these findings strongly suggest that inappropriate expression of PDZK4 might play an important role in the proliferation of SS cells and that the gene might be a suitable molecular target for designing of novel drugs to treat SS patients.
T Nakamura, Y Furukawa, H Nakagawa, T Tsunoda, H Ohigashi, K Murata, O Ishikawa, K Ohgaki, N Kashimura, M Miyamoto, S Hirano, S Kondo, H Katoh, Y Nakamura and Toyomasa Katagiri : Genome-wide cDNA microarray analysis of gene-expression profiles in pancreatic cancers using populations of tumor cells and normal ductal epithelial cells selected for purity by laser microdissection, Oncogene, Vol.23, No.13, 2385-400, 2004.
(要約)
To characterize molecular mechanism involved in pancreatic carcinogenesis, we analysed gene-expression profiles of 18 pancreatic tumors using a cDNA microarray representing 23,040 genes. As pancreatic ductal adenocarcinomas usually contain a low proportion of cancer cells in the tumor mass, we prepared 95% pure populations of pancreatic cancer cells by means of laser microbeam microdissection, and compared their expression profiles to those of similarly purified, normal pancreatic ductal cells. We identified 260 genes that were commonly upregulated and 346 genes that were downregulated in pancreatic cancer cells. Because of the high degree of purity in the cell populations, a large proportion of genes that we detected as upregulated or downregulated in pancreatic cancers were different from those reported in previous studies. Comparison of clinicopathological parameters with the expression profiles indicated that altered expression of 76 genes was associated with lymph-node metastasis and that of 168 genes with liver metastasis. In addition, expression levels of 30 genes were related to the recurrence of disease. These genome-wide expression profiles should provide useful information for finding candidate genes whose products might serve as specific tumor markers and/or as molecular targets for treatment of patients with pancreatic cancer.
K Ochi, Y Daigo, Toyomasa Katagiri, S Nagayama, T Tsunoda, A Myoui, N Naka, N Araki, I Kudawara, M Ieguchi, Y Toyama, J Toguchida, H Yoshikawa and Y Nakamura : Prediction of response to neoadjuvant chemotherapy for osteosarcoma by gene-expression profiles, International Journal of Oncology, Vol.24, No.3, 647-655, 2004.
(要約)
To establish a method for predicting the response to chemotherapy for osteosarcoma (OS), we performed expression profile analysis using cDNA microarray consisting of 23,040 genes. Hierarchical clustering based on the expression profiles of 19 biopsy samples of OS demonstrated two major clusters, one of which consisted exclusively of typical OS, i.e. conventional central OS in long bone of patients in the second decade. A set of genes was identified to characterize this subgroup, some of which have previously indicated some relation to carcinogenesis. Thirteen of the 19 patients were treated with an identical protocol of chemotherapy containing doxorubicin, cis-platinum and ifosfamide, and histological examination of resected specimens after operation classified 6 cases as responder and 7 as non-responder. A comparison of expression profiles of these two groups identified 60 genes whose expression levels were likely to be correlated with the response to chemotherapy (P<0.008). A drug response scoring (DRS) system was developed based on the expression levels of these genes, which proved to be applicable to predict the response to chemotherapy irrespective for the subclassification of OS. The reliability of the DRS system was further confirmed by testing additional 5 OS cases. These results indicated that scoring system based on gene-expression profiles might be useful to predict the response to chemotherapy for OS.
M Li, YM Lin, S Hasegawa, T Shimokawa, K Murata, M Kameyama, O Ishikawa, Toyomasa Katagiri, T Tsunoda, Y Nakamura and Y Furukawa : Genes associated with liver metastasis of colon cancer, identified by genome-wide cDNA microarray, International Journal of Oncology, Vol.24, No.2, 305-312, 2004.
(要約)
To uncover mechanisms underlying progression of colorectal carcinogenesis and to identify genes associated with liver metastasis, we analyzed expression profiles of 14 primary colorectal cancers (CRCs) with liver metastases, and compared them with profiles of 11 non-metastatic carcinomas and those of 9 adenomas of the colon. A hierarchical cluster analysis using data from a cDNA microarray containing 23,040 genes indicated that the cancers with metastasis had different expression profiles from those without metastasis, although a number of genes were commonly up-regulated in primary cancers of both categories. We documented 54 genes that were frequently up-regulated and 375 that were frequently down-regulated in primary tumors with metastases to liver, but not in tumors without metastasis. Subsequent quantitative PCR experiments confirmed that PRDX4, CKS2, MAGED2, and an EST (GenBank accession number BF696304) were expressed at significantly higher levels in tumors with metastasis. These data should contribute to a better understanding of the progression of colorectal tumors, and facilitate prediction of their metastatic potential.
K Okada, Toyomasa Katagiri, K Okada, Y Mizutani, Y Suzuki, M Kamada, T Fujioka, T Shuin, T Miki and Y Nakamura : Analysis of gene-expression profiles in testicular seminomas using a genome-wide cDNA microarray, International Journal of Oncology, Vol.23, No.6, 1615-1635, 2003.
(要約)
To identify new diagnostic markers for testicular germ cell tumors (TGCTs), including seminomas, as well as potential targets of new drugs for treating the disease, we compared gene-expression profiles of cancer cells from 13 seminomas with normal human testis using laser-capture microdissection and a cDNA microarray representing 23,040 genes. We identified 347 genes that were commonly up-regulated in seminoma cells. The functions of 227 were known to some extent; the remaining 120 included 55 ESTs. On the list were cyclin D2 (CCND2), prostate cancer over-expressed gene 1 (POV1), and junction plakoglobin (JUP), all of which were already known to be over-expressed in seminomas. On the other hand, our protocol selected 593 genes as being commonly down-regulated in seminoma cells. That list included 340 functionally characterized genes; the other 253 included 131 ESTs. To confirm the expression data, we performed semi-quantitative RT-PCR experiments with nine highly up-regulated genes, and the results supported those of our microarray analysis. The information provided here should prove useful for identifying genes whose products might serve as molecular targets for treatment of TGCTs.
C Suzuki, Y Daigo, T Kikuchi, Toyomasa Katagiri and Y Nakamura : Identification of COX17 as a therapeutic target for non-small cell lung cancer, Cancer Research, Vol.63, No.21, 7038-7041, 2003.
(要約)
We have been investigating gene expression profiles in non-small cell lung cancers (NSCLCs) to identify molecules involved in pulmonary carcinogenesis and select which genes or gene products might be useful as diagnostic markers or targets for new molecular therapies. Here we report evidence that the cytochrome c oxidase (CCO) assembly protein COX17 is a potential molecular target for treatment of lung cancers. By semiquantitative reverse transcription-PCR, we documented increased expression of COX17 in all of 8 primary NSCLCs and in 11 of 15 NSCLC cell lines examined, by comparison with normal lung tissue. Treatment of NSCLC cells with antisense S-oligonucleotides or vector-based small interfering RNAs of COX17 suppressed expression of COX17 and also the activity of CCO, and suppressed growth of the cancer cells. Because our data imply that up-regulation of COX17 function and increased CCO activity are frequent features of lung carcinogenesis, we suggest that selective suppression of components of the CCO complex might hold promise for development of a new strategy for treating lung cancers.
(キーワード)
Amino Acid Sequence / Animals / COS Cells / Carcinoma, Non-Small-Cell Lung / Carrier Proteins / Cation Transport Proteins / Cell Line, Tumor / Cercopithecus aethiops / Electron Transport Complex IV / Gene Expression / Humans / Lung Neoplasms / Molecular Sequence Data / Oligonucleotide Array Sequence Analysis / Oligonucleotides, Antisense / RNA, Small Interfering
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 14612491
Y Kaneta, Y Kagami, T Tsunoda, R Ohno, Y Nakamura and Toyomasa Katagiri : Genome-wide analysis of gene-expression profiles in chronic myeloid leukemia cells using a cDNA microarray, International Journal of Oncology, Vol.23, No.3, 681-691, 2003.
(要約)
To characterize molecular mechanisms operating in chronic myeloid leukemia (CML) cells with a view toward development of novel therapeutic targets, we analyzed gene-expression profiles of cancer cells from 27 CML patients using a cDNA microarray representing 23,040 human genes. By comparing expression patterns of CML with those of normal cells, we identified 150 genes that were commonly highly up-regulated in CML cells. In addition to 54 genes (34 of them ESTs) whose functions are currently unknown, the up-regulated elements included genes encoding cell-cycle regulators, transcriptional activators, transcriptional factors, and protein kinases as well as proteins already known to be induced in CML, such as some hemoglobins, haptoglobin (HP1), and matrix metalloproteinase 9 (MMP-9), a protein involved in tissue remodeling and tumor invasion. On the other hand, our protocol selected 106 genes, including 13 of unknown function, as being commonly significantly down-regulated in all phases of CML. The results of semiquantitative RT-PCR experiments with 11 representatives of the up-regulated group supported the reliability of our microarray analysis. These data should provide useful information for finding candidate genes whose products might serve as molecular targets for treatment of CML patients.
H Zembutsu, Y Ohnishi, Y Daigo, Toyomasa Katagiri, T Kikuchi, S Kakiuchi, C Nishime, K Hirata and Y Nakamura : Gene-expression profiles of human tumor xenografts in nude mice treated orally with the EGFR tyrosine kinase inhibitor ZD1839, International Journal of Oncology, Vol.23, No.1, 29-39, 2003.
(要約)
To date, no single or multiple molecular markers have been successful in predicting sensitivity of individual patients to anti-cancer drugs. As the nature of a specific cancer is considered to be defined by the proteins being expressed in the tumor cells, systematic analysis of gene-expression profiles may provide information reflecting sensitivity of a given tumor to certain drugs. Recent progress in genome technology has enabled us to examine expression profiles of thousands of genes in a single experiment. We used this approach to examine 13 xenografts of human tumors implanted into nude mice for sensitivity to an orally active, selective epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), ZD1839 (Iressa). To identify genes that might be associated with sensitivity to this drug we used a cDNA microarray representing 23,040 genes to analyze expression profiles of the 13 xenografts and identified 114 genes whose expression levels correlated significantly with sensitivity of the tumors to ZD1839. We then investigated alteration of expression profiles in response to the ZD1839 treatment in four non-small cell lung cancer (NSCLC) xenografts, of which two (LC6 and LC11) were sensitive and the other two (Lu116 and L27) were resistant to this EGFR-TKI. Systematic analysis of expression at various time points during oral treatment for 14 days, compared with corresponding untreated samples, identified a set of genes whose expression levels changed in the two sensitive tumors but not in the two resistant tumors. The data obtained here should provide useful information on the molecular mechanism underlying clinical responses to EGFR-TKIs, aid the development of novel therapies for lung cancer, and potentially identify predictive molecular markers for sensitivity to ZD1839.
T Nakatsura, Y Yoshitake, S Senju, M Monji, H Komori, Y Motomura, S Hosaka, T Beppu, T Ishiko, H Kamohara, H Ashihara, Toyomasa Katagiri, Y Furukawa, S Fujiyama, M Ogawa, Y Nakamura and Y Nishimura : Glypican-3, overexpressed specifically in human hepatocellular carcinoma, is a novel tumor marker, Biochemical and Biophysical Research Communications, Vol.306, No.1, 16-25, 2003.
(要約)
With the global pandemic of hepatitis B and C infections, the incidence of Hepatocellular carcinoma (HCC) is rapidly increasing world wide. We identified glypican-3 (GPC3), a novel oncofetal gene over-expressed specifically in human HCC, as based on data of cDNA microarrays. As GPC3 is a GPI-anchored membrane protein and could be secreted, we attempted to detect secreted GPC3 protein in sera from HCC patients using Western blotting and ELISA. GPC3 protein was positive in sera of 40.0% (16/40) of HCC patients, and negative in sera from subjects with liver cirrhosis (LC) (0/13), chronic hepatitis (CH) (0/34), and healthy donors (0/60). All subjects were Japanese. Although 12 of 40 HCC patients were negative for both alpha-fetoprotein (AFP) and PIVKA-II well known tumor markers of HCC, four of these were GPC3-positive in the sera. We also observed vanishing GPC3 protein in the sera of three patients after the surgical treatment for HCC. On the other hand, immunohistochemical analysis revealed that HCC expressed GPC3 protein in all 14 HCC patients tested. In conclusion, GPC3, as defined in this study was shown to be a useful tumor marker for cancer-diagnosis for large numbers of patients with HCC.
Takefumi Kikuchi, Yataro Daigo, Toyomasa Katagiri, Tatsuhiko Tsunoda, Koichi Okada, Soji Kakiuchi, Hitoshi Zembutsu, Yoichi Furukawa, Masafumi Kawamura, Koichi Kobayashi, Kohzoh Imai and Yusuke Nakamura : Expression profiles of non-small cell lung cancers on cDNA microarrays: identification of genes for prediction of lymph-node metastasis and sensitivity to anti-cancer drugs, Oncogene, Vol.22, No.14, 2192-205, 2003.
(要約)
To investigate genes involved in pulmonary carcinogenesis and those related to sensitivity of nonsmall cell lung cancers (NSCLCs) to therapeutic drugs, we performed cDNA microarray analysis of 37 NSCLCs after laser-capture microdissection of cancer cells from primary tumors. A clustering algorithm applied to the expression data easily distinguished two major histological types of non-small cell lung cancer, adenocarcinoma and squamous cell carcinoma. Subsequent analysis of the 18 adenocarcinomas identified 40 genes whose expression levels could separate cases with lymph-node metastasis from those without metastasis. In addition, we compared the expression data with measurements of the sensitivity of surgically dissected NSCLC specimens to six anti-cancer drugs (docetaxel, paclitaxel, irinotecan, cisplatin, gemcitabine, and vinorelbine), as measured by the CD-DST (collagen gel droplet embedded culture-drug sensitivity test) method. We found significant associations between expression levels of dozens of genes and chemosensitivity of NSCLCs. Our results provide valuable information for eventually identifying predictive markers and novel therapeutic target molecules for this type of cancer.
T Arimoto, Toyomasa Katagiri, K Oda, T Tsunoda, T Yasugi, Y Osuga, H Yoshikawa, O Nishii, T Yano, Y Taketani and Y Nakamura : Genome-wide cDNA microarray analysis of gene-expression profiles involved in ovarian endometriosis, International Journal of Oncology, Vol.22, No.3, 551-560, 2003.
(要約)
Using a cDNA microarray consisting of 23,040 genes, we analyzed gene-expression profiles of ovarian endometrial cysts from 23 patients in order to identify genes involved in endometriosis. By comparing expression patterns between endometriotic tissues and corresponding eutopic endometria, we identified 15 genes that were commonly up-regulated in the endometrial cysts during both proliferative and secretory phases of the menstrual cycle, 42 that were up-regulated only in the proliferative phase, and 40 that were up-regulated only in the secretory phase. The up-regulated elements included genes encoding some HLA antigens, complement factors, ribosomal proteins, and TGFBI. On the other hand, 337 genes were commonly down-regulated throughout the menstrual cycle, 144 only in the proliferative phase, and 835 only in the secretory phase. The down-regulated elements included the tumor suppressor TP53, genes related to apoptosis such as GADD34, GADD45A, GADD45B and PIG11, and the gene encoding OVGP1, a protein involved in maintenance of early pregnancy. Semi-quantitative RT-PCR experiments supported the results of our microarray analysis. These data should provide useful information for finding candidate genes whose products might serve as molecular targets for diagnosis or treatment of endometriosis.
K Ochi, Y Daigo, Toyomasa Katagiri, A Saito-Hisaminato, T Tsunoda, Y Toyama, H Matsumoto and Y Nakamura : Expression profiles of two types of human knee-joint cartilage, Journal of Human Genetics, Vol.48, No.4, 177-182, 2003.
(要約)
We have performed a comprehensive analysis of gene-expression profiles in human articular cartilage (hyaline cartilage) and meniscus (fibrocartilage) by means of a cDNA microarray consisting of 23,040 human genes. Comparing the profiles of the two types of cartilage with those of 29 other normal human tissues identified 24 genes that were specifically expressed in both cartilaginous tissues; these genes might be involved in maintaining phenotypes common to cartilage. We also compared the cartilage profiles with gene expression in human mesenchymal stem cells (hMSC), and detected 22 genes that were differentially expressed in cells representing the two cartilaginous lineages, 11 specific to each type, which could serve as markers for predicting the direction of chondrocyte differentiation. Our data should also provide useful information about regeneration of cartilage, especially in support of efforts to identify cartilage-specific molecules as potential agents for therapeutic approaches to joint repair.
Satoko Abe, Toyomasa Katagiri, Akihiko Saito-Hisaminato, Shin-ichi Usami, Yasuhiro Inoue, Tatsuhiko Tsunoda and Yusuke Nakamura : Identification of CRYM as a candidate responsible for nonsyndromic deafness, through cDNA microarray analysis of human cochlear and vestibular tissues., American Journal of Human Genetics, Vol.72, No.1, 73-82, 2003.
(要約)
Through cDNA microarray analysis of gene expression in human cochlea and vestibule, we detected strong expression of mu-crystallin (CRYM; also known as "NADP-regulated thyroid hormone-binding protein") only in these inner-ear tissues. In a subsequent search for mutations of CRYM, among 192 patients with nonsyndromic deafness, we identified two mutations at the C-terminus; one was a de novo change (X315Y) in a patient with unaffected parents, and the other was a missense mutation (K314T) that segregated dominantly in the proband's family. When the mutated proteins were expressed in COS-7 cells, their subcellular localizations were different from that of the normal protein: the X315Y mutant showed vacuolated distribution in the cytoplasm, and the K314T mutant localized in perinuclear areas, whereas normal protein was distributed homogeneously in the cytoplasm. Aberrant intracellular localization of the mutated proteins might cause dysfunction of the CRYM product and result in hearing impairment. In situ hybridization analysis using mouse tissues indicated its expression in the lateral region of the spiral ligament and the fibrocytes of the spiral limbus, implying its possible involvement in the potassium-ion recycling system. Our results strongly implicate CRYM in normal auditory function and identify it as one of the genes that can be responsible for nonsyndromic deafness.
(キーワード)
Amino Acid Sequence / Animals / Base Sequence / COS Cells / Cochlea / Crystallins / Cytoplasm / DNA Mutational Analysis / Deafness / Female / Gene Expression Profiling / Humans / In Situ Hybridization / Male / Molecular Sequence Data / Mutation / Oligonucleotide Array Sequence Analysis / Pedigree / RNA, Messenger / Reproducibility of Results / Vestibule, Labyrinth
S Hasegawa, Y Furukawa, M Li, S Satoh, T Kato, T Watanabe, Toyomasa Katagiri, T Tsunoda, Y Yamaoka and Y. Nakamura : Genome-wide analysis of gene expression in intestinal-type gastric cancers using a complementary DNA microarray representing 23,040 genes, Cancer Research, Vol.62, No.23, 7012-7017, 2002.
(要約)
To shed light on mechanisms that underlie development and/or progression of intestinal-type gastric cancer, we compared expression profiles of cancer cells obtained by laser-capture microdissection of 20 intestinal-type gastric tumors with expression of genes in corresponding noncancerous mucosae, by a cDNA microarray consisting of 23,040 genes. We identified 61 genes that were commonly up-regulated and 63 that were commonly down-regulated in the cancer tissues. Altered expression of 12 of those genes was associated with lymph node metastasis. A "predictive score," based on expression profiles of five of the genes that were able to distinguish tumors with metastasis from node-negative tumors in our panel, correctly diagnosed the lymph node status of nine additional gastric cancers. This genome-wide information contributes to an improved understanding of molecular changes during the development of intestinal-type gastric cancers. It may help clinicians predict metastasis to lymph nodes and assist researchers in identifying novel therapeutic targets for this type of cancer.
S Nagamaya, Toyomasa Katagiri, T Tsunoda, T Hosaka, Y Nakashima, N Araki, K Kusuzaki, T Nakayama, T Tsuboyama, T Nakamura, M Imamura, Y Nakamura and J. Toguchida : Genome-wide analysis of gene expression in synovial sarcomas using a cDNA microarray, Cancer Research, Vol.62, No.20, 5859-5866, 2002.
(要約)
Among a histologically heterogeneous group of soft tissue sarcomas, synovial sarcoma (SS) is regarded as a "miscellaneous" entity of uncertain origin. Although recent molecular analysis has disclosed involvement of a specific chromosomal translocation in the pathogenesis of SS, its genetic features remain largely unclear. In the work reported here we examined genome-wide gene expression profiles of 13 SS cases and 34 other spindle-cell sarcoma cases by cDNA microarray consisting of 23,040 genes. A hierarchical clustering analysis grouped SS and malignant peripheral nerve sheath tumor into the same category, and these two types of tumor shared expression patterns of numerous genes relating to neural differentiation. Several genes were up-regulated in almost all SS cases, and the presumed functions of known genes among them were related to migration or differentiation of neural crest cells, suggesting the possibility of neuroectodermal origin of SS. Moreover, we identified a set of genes that divided SS cases into two putative subclasses, a feature that may shed light on novel biological aspects of SS in addition to those having to do with epithelial differentiation. These data have provided clues for understanding the origin and tumorigenesis of SS.
M Brown, H Nicolai, K Howe, Toyomasa Katagiri, E Lalani, K Simpson, N Manning, A Deans, P Chen, KK Khanna, MR Wati, B Griffiths, C Xu, G Stamp and E. Solomon : Expression of a truncated Brca1 protein delays lactational mammary development in transgenic mice, Transgenic Research, Vol.11, No.5, 467-478, 2002.
(要約)
To address the hypothesis that certain disease-associated mutants of the breast-ovarian cancer susceptibility gene BRCA1 have biological activity in vivo, we have expressed a truncated Brca1 protein (trBrca1) in cell-lines and in the mammary gland of transgenic mice. Immunofluorescent analysis of transfected cell-lines indicates that trBRCA1 is a stable protein and that it is localized in the cell cytoplasm. Functional analysis of these cell-lines indicates that expression of trBRCA1 confers an increased radiosensitivity phenotype on mammary epithelial cells, consistent with abrogation of the BRCA1 pathway. MMTV-trBrca1 transgenic mice from two independent lines displayed a delay in lactational mammary gland development, as demonstrated by altered histological profiles of lobuloalveolar structures. Cellular and molecular analyses indicate that this phenotype results from a defect in differentiation, rather than altered rates of proliferation or apoptosis. The results presented in this paper are consistent with trBrca1 possessing dominant-negative activity and playing an important role in regulating normal mammary development. They may also have implications for germline carriers of BRCA1 mutations.
J Okutsu, T Tsunoda, Y Kaneta, Toyomasa Katagiri, O Kitahara, H Zembutsu, R Yanagawa, S Miyawaki, K Kuriyama, N Kubota, Y Kimura, K Kubo, F Yagasaki, T Higa, H Taguchi, T Tobita, H Akiyama, A Takeshita, Y Wang, T Motoji, R Ohno and Y Nakamura : Prediction of chemosensitivity for patients with acute myeloid leukemia, according to expression levels of 28 genes selected by genome-wide complementary DNA microarray analysis, Molecular Cancer Therapeutics, Vol.1, No.12, 1035-1042, 2002.
(要約)
To identify genes involved in the sensitivity of acute myeloid leukemia (AML) cells to chemotherapy, we monitored gene-expression profiles of cancer cells from 76 AML patients using a cDNA microarray consisting of 23,040 genes. We identified 63 genes that were commonly overexpressed and 372 genes suppressed in AML. Because these genes represent key molecules for disclosing the molecular mechanisms of AML, they may be potential targets for drug development. We also found 28 that revealed different expression levels between good and poor responders to chemotherapy and appeared to be associated with chemosensitivity. On that basis, we developed a "Drug Response Scoring" system that was correlated well with individual sensitivity to an anticancer drug regimen. Among the 44 cases with positive drug-response scores by our definition, 40 achieved complete remission after treatment, whereas the only 3 of the 20 cases with negative scores responded well to the treatment. An ability to predict chemosensitivity should eventually lead to achievement of our goal of "personalized therapy."
Y Kaneta, Y Kagami, Toyomasa Katagiri, T Tsunoda, I Jin-nai, H Taguchi, H Hirai, K Ohnishi, T Ueda, N Emi, A Tomida, T Tsuruo, Y Nakamura and R. Ohno : Prediction of sensitivity to STI571 among chronic myeroid leukemia patients by genome-wide cDNA microarray analysis, Japanese Journal of Cancer Research, Vol.93, No.8, 849-856, 2002.
(要約)
One of the most critical issues to be solved in regard to cancer chemotherapy is the establishment of ways to predict the efficacy of anti-cancer drugs for individual patients. To develop a prediction system based on expression of specific genes, we analyzed expression profiles of mononuclear cells from 18 chronic myeloid leukemia (CML) patients who were treated with the tyrosine kinase inhibitor STI571. cDNA microarrays representing 23 040 genes identified 79 genes that were expressed differentially between responders and non-responders to STI571. On the basis of the expression patterns of 15 or 30 of these genes among the patients, we developed a "Prediction Score" system that could clearly separate the responder group from the non-responder group. Verification of this system using four additional ("test") cases succeeded in predicting the response of each of those four patients to the drug. These results provide the first evidence that gene-expression profiles can predict sensitivity of CML cells to STI571, and may eventually lead to the achievement of "personalized therapy" for this disease.
K Miura, ED Bowman, R Simon, AC Peng, AI Robles, RT Jones, Toyomasa Katagiri, P He, H Mizukami, L Charboneau, T Kikuchi, LA Liotta, Y Nakamura and CC. Harris : Laser capture microdissection and microarray expression analysis of lung adenocarcinoma reveals tobacco smoking and prognosis-related molecular profiles., Cancer Research, Vol.62, No.11, 3244-3250, 2002.
(要約)
Recent expression profile analyses revealed that lung adenocarcinomas can be divided into several subgroups with diverse pathological features. Because cellular heterogeneity of tumors can confound these analyses, we used laser capture microdissection and microarray expression analysis to characterize the molecular profiles of lung adenocarcinomas. We found 45 genes delineating smokers and nonsmokers that were located at chromosomal loci frequently altered in non-small cell lung cancers, and 27 genes, which were differentially expressed between survivors and nonsurvivors 5 years after surgery. These results are consistent with the hypothesis that the abnormal expression of genes involved in maintaining the mitotic spindle checkpoint and genomic stability, e.g., hBUB3, hZW10, and APC2, contribute to the molecular pathogenesis and tumor progression of tobacco smoke-induced adenocarcinoma of the lung.
Osamu Kitahara, Toyomasa Katagiri, Tatsuhiko Tsunoda, Yoko Harima and Yusuke Nakamura : Classification of sensitivity or resistance of cervical cancers to ionizing radiation according to expression profiles of 62 genes selected by cDNA microarray analysis., Neoplasia, Vol.4, No.4, 295-303, 2002.
(要約)
To identify a set of genes related to radiosensitivity of cervical squamous cell carcinomas and to establish a predictive method, we compared expression profiles of 9 radiosensitive and 10 radioresistant tumors obtained by biopsy before treatment, on a cDNA microarray consisting of 23,040 human genes. We identified 121 genes whose expression was significantly greater in radiosensitive cells than in radioresistant cells, and 50 genes that showed higher levels of expression in radioresistant cells than in radiosensitive cells. Some of these genes had already known to be associated with the radiation response, such as aldehyde dehydrogenase 1 (ALDH1) and X-ray repair cross-complementing 5 (XRCC5) (P<.05, Mann-Whitney test). The validity of the total of 171 genes as radiosensitivity related genes were certified by permutation test (P<.05). Furthermore, we selected 62 genes on the basis of a clustering analysis, and confirmed the validity of these genes with cross-validation test. The cross-validation test also indicates the possibility of making prediction of radiosensitivity for discriminating radiation-sensitive from radiation resistant biopsy samples by predicting score (PS) values calculated from expression values of 62 genes in 19 samples, because the prediction successfully and unequivocally discriminated the radiosensitive phenotype from the radioresistant phenotype in our test panel of 19 cervical carcinomas. The extensive list of genes identified in these experiments provides a large body of potentially valuable information for studying the mechanism(s) of radiosensitivity, and selected 62 genes opens the possibility of providing appropriate and effective radiotherapy to cancer patients.
A Saito-Hisaminato, Toyomasa Katagiri, S Kakiuchi, T Nakamura, T Tsunoda and Y Nakamura : Genome-wide profiling of gene expression in 29 normal human tissues with a cDNA microarray., DNA Research, Vol.9, No.2, 35-45, 2002.
(要約)
We have performed a comprehensive analysis of the expression profiles in 25 adult and 4 fetal human tissues by means of a cDNA microarray consisting of 23,040 human genes. This study revealed a number of genes that were expressed specifically in each of those tissues. Among the 29 tissues examined, 4,080 genes were highly expressed (at least a five-fold expression ratio) in one or only a few tissues and 1,163 of those were expressed exclusively (more than a ten-fold higher expression ratio) in a particular tissue. Expression of some of the genes in the latter category was confirmed by northern analysis. A hierarchical clustering analysis of gene-expression profiles in nerve tissues (adult brain, fetal brain, and spinal cord), lymphoid tissues (bone marrow, thymus, spleen, and lymph node), muscle tissues (heart and skeletal muscle), or adipose tissues (mesenteric adipose and mammary gland) identified a set of genes that were commonly expressed among related tissues. These data should provide useful information for medical research, especially for efforts to identify tissue-specific molecules as potential targets of novel drugs to treat human diseases.
Shingo Dan, Tatsuhiko Tsunoda, Osamu Kitahara, Rempei Yanagawa, Hitoshi Zembutsu, Toyomasa Katagiri, Kanami Yamazaki, Yusuke Nakamura and Takao Yamori : An Integrated Database of Chemosensitivity to 55 Anticancer Drugs and Gene Expression Profiles of 39 Human Cancer Cell Lines, Cancer Research, Vol.62, No.4, 1139-47, 2002.
(要約)
To explore genes that determine the sensitivity of cancer cells to anticancer drugs, we investigated using cDNA microarrays the expression of 9216 genes in 39 human cancer cell lines pharmacologically characterized on treatment with various anticancer drugs. A bioinformatical approach was then exploited to identify genes related to anticancer drug sensitivity. An integrated database of gene expression and drug sensitivity profiles was constructed and used to identify genes with expression patterns that showed significant correlation to patterns of drug responsiveness. As a result, sets of genes were extracted for each of the 55 anticancer drugs examined. Whereas some genes commonly correlated with various classes of anticancer drugs, other genes correlated only with specific drugs with similar mechanisms of action. This latter group of genes may reflect the efficacy of each class of drugs. Therefore, the integrated database approach of gene expression and chemosensitivity profiles may be useful in the development of systems to predict drug efficacies of cancer cells by examining the expression levels of particular genes.
H Zembutsu, Y Ohnishi, T Tsunoda, Y Furukawa, Toyomasa Katagiri, Y Ueyama, N Tamaoki, T Nomura, O Kitahara, R Yanagawa, K Hirata and Y. Nakamura : Genome-wide cDNA microarray screening to correlate gene expression profiles with sensitivity of 85 human cancer xenografts to anticancer drugs., Cancer Research, Vol.62, No.2, 518-527, 2002.
(要約)
One of the most critical issues to be solved in regard to cancer chemotherapy is the need to establish a method for predicting efficacy or toxicity of anticancer drugs for individual patients. To identify genes that might be associated with chemosensitivity, we used a cDNA microarray representing 23,040 genes to analyze expression profiles in a panel of 85 cancer xenografts derived from nine human organs. The xenografts, implanted into nude mice, were examined for sensitivity to nine anticancer drugs (5-fluorouracil, 3-[(4-amino-2-methyl-5-pyrimidinyl)methyl]-1-(2-chloroethyl)-1-nitrosourea hydrochloride, adriamycin, cyclophosphamide, cisplatin, mitomycin C, methotrexate, vincristine, and vinblastine). Comparison of the gene expression profiles of the tumors with sensitivities to each drug identified 1,578 genes whose expression levels correlated significantly with chemosensitivity; 333 of those genes showed significant correlation with two or more drugs, and 32 correlated with six or seven drugs. These data should contribute useful information for identifying predictive markers for drug sensitivity that may eventually provide "personalized chemotherapy" for individual patients, as well as for development of novel drugs to overcome acquired resistance of tumor cells to chemical agents.
(キーワード)
Animals / Antineoplastic Agents / Cluster Analysis / Gene Expression Profiling / Humans / Mice / Mice, Inbred BALB C / Mice, Nude / Neoplasms / Oligonucleotide Array Sequence Analysis / Predictive Value of Tests / Xenograft Model Antitumor Assays
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 11809704
K Maeda, S Matsuhashi, K Tabuchi, T Watanabe, Toyomasa Katagiri, M Oyasu, N Saito and S. Kuroda : Brain specific human genes, NELL1 and NELL2, are predominantly expressed in neuroblastoma and other embryonal neuroepithelial tumors, Neurologia Medico-Chirurgica, Vol.41, No.12, 582-589, 2001.
(要約)
NELL1 and NELL2 encode cysteine-rich amino acid sequences including six epidermal growth factor-like motifs, which contain signal peptides at the N-terminals. The deduced amino acid sequences of both genes are 55% identical and their cysteine stretch structures are conserved. NELL1 is expressed in the brain and kidney, whereas NELL2 is expressed specifically in the brain. The cell lineage expressing NELLs in the nervous system was investigated in established cell lines and central nervous system tumor tissues obtained from patients by Northern blot and reverse transcriptase-polymerase chain reaction analyses. NELL1 and NELL2 were predominantly expressed in neuroblastoma cell lines and little expressed in glioblastoma cell lines. NELL1 and NELL2 were also expressed in central neurocytoma, medulloblastoma, and some astrocytic tumors. Immunohistochemical analysis revealed that NELL2 protein was localized in the cytoplasm of neurons. These results suggest that NELL2 is predominantly expressed in the neuronal cell lineage in the human nervous system. NELL1 is expressed mainly in tumors in the neuronal cell lineage.
M Futamura, H Arakawa, K Matsuda, Toyomasa Katagiri, S Saji, Y Miki and Y. Nakamura : Potential role of BRCA2 in a mitotic checkpoint after phosphorylation by hBUBR1, Cancer Research, Vol.60, No.6, 1531-1535, 2000.
(要約)
BRCA2, a gene responsible for inherited susceptibility to breast cancer in a number of families, is thought to be critical for replication and repair of DNA during S-phase. To elucidate the physiological functions of BRCA2, we used a yeast two-hybrid system to screen for proteins that could associate with BRCA2. Here we report interaction of BRCA2 with a mitotic checkpoint protein, hBUBR1, and its phosphorylation by hBUBR1 in vitro. After cotransfection of BRCA2 and hBUBR1 expression vectors into the COS7 cell line, both proteins were stained together in the nuclei of cells whose spindle fibers were disrupted, but not in undamaged cells. Treatment with vincristine, which disrupts microtubules, significantly increased expression of both hBUBR1 and BRCA2 in the MCF7 cells. The results suggest that BRCA2 protein might be involved in a mitotic checkpoint in vivo after it has been phosphorylated by hBUBR1.
M Kato, K Yano, F Matsuo, H Saito, Toyomasa Katagiri, H Kurumizaka, M Yoshimoto, F Kasumi, F Akiyama, G Sakamoto, H Nagawa, Y Nakamura and Y Miki : Identification of Rad51 alteration in patients with bilateral breast cancer, Journal of Human Genetics, Vol.45, No.3, 133-137, 2000.
(要約)
The human Rad51 gene, HsRAD51, is a homolog of RecA of Escherichia coli and functions in recombination and DNA repair. BRCA1 and BRCA2 proteins form a complex with Rad51, and these genes are thought to participate in a common DNA damage response pathway associated with the activation of homologous recombination and double-strand break repair. Additionally, we have shown that the pattern of northern blot analysis of the RadS gene is closely similar to those of the BRCA1 and BRCA2 genes. It is therefore possible that alterations of the Rad51 gene may be involved in the development of hereditary breast cancer. To investigate this possibility, we screened Japanese patients with hereditary breast cancer for Rad51 mutations and found a single alteration in exon 6. This was determined to be present in the germline in two patients with bilateral breast cancer, one with synchronous bilateral breast cancer and the other with synchronous bilateral multiple breast cancer. In both patients, blood DNAs showed a G-to-A transition in the second nucleotide of codon 150, which results in the substitution of glutamine for arginine. As this alteration was not present in any patients with breast or colon cancer examined, we assume that this missense alteration is likely to be a disease-causing mutation.
(キーワード)
Blotting, Northern / Breast Neoplasms / DNA Mutational Analysis / DNA-Binding Proteins / Exons / Family Health / Female / Humans / Japan / Point Mutation / Polymerase Chain Reaction / Polymorphism, Single-Stranded Conformational / Rad51 Recombinase
M Emi, M Yoshimoto, T Sato, S Matsumoto, Y Utada, I Ito, K Minobe, T Iwase, Toyomasa Katagiri, K Bando, F Akiyama, Y Harada, K Fukino, G Sakamoto, M Matsushima, A Iida, T Tada, H Saito, Y Miki, F Kasumi and Y Nakamura : Allelic loss at 1p34, 13q12, 17p13.3, and 17q21.1 correlates with poor postoperative prognosis in breast cancer., Genes, Chromosomes & Cancer, Vol.26, No.2, 134-141, 1999.
(要約)
Allelic losses of tumor suppressor genes (TSGs), or the chromosomal regions harboring them, in tumor DNA may become useful postoperative prognostic indicators. To examine whether specific allelic losses might correlate with postoperative survival in a 5-year prospective follow-up, we tested tumors from a cohort of 264 breast cancer patients for allelic losses of 18 microsatellite markers representing either a known TSG or a region where genetic alterations are frequent in breast tumors. Patients whose tumors had lost an allele at 1p34, 13q12, 17p13.3, or 17q21.1 had significantly higher risks of postoperative mortality than those whose tumors retained both alleles at those loci (at 1p34, a 5-year mortality rate of 29% among patients with losses vs. 7% with retentions, P = 0. 0008; at 13q12, 31% vs. 10%, P = 0.0062; at 17p13.3, 24% vs. 13%, P = 0.026; and at 17q21.1, 31% vs. 13%, P = 0.0047). Furthermore, combined losses at 13q12 and 17p13.3 increased the predicted postoperative mortality risks by a factor of 9.6 (5-year mortality rate of 42% vs. 5% with retentions, P = 0.0001), and combined losses at 1p34 and 17p13.3 raised the predicted postoperative mortality risks by a factor of 8.6 (27% vs. 3%, P = 0.0064). We conclude that allelic losses at these loci can serve as negative prognostic indicators to guide postoperative management of patients. Genes Chromosomes Cancer 26:134-141, 1999.
(キーワード)
Adult / Aged / Aged, 80 and over / Breast Neoplasms / Chromosomes, Human / Chromosomes, Human, Pair 1 / Chromosomes, Human, Pair 13 / Chromosomes, Human, Pair 17 / Female / Humans / Loss of Heterozygosity / Middle Aged / Postoperative Period / Prognosis / Survival Rate
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 10469451
Toyomasa Katagiri, M Futamura and Y. Nakamura : A Gln/Arg polymorphism at codon 349 of the hBUBR1 gene., Journal of Human Genetics, Vol.44, No.2, 131-2, 1999.
(要約)
We found a glutamine/arginine polymorphism at codon 349 of the hBUBR1 gene, encoding a protein kinase required for spindle assembly checkpoint function. The observed heterozygosity was estimated to be 45% in the Japanese population. This polymorphism may be helpful for genetic studies of many cancer types in which chromosomal instability is observed.
M Emi, M Matsushima, Toyomasa Katagiri, T Yokota, T Nakata, F Kasumi, Y Miki, MH Skolnick and Y. Nakamura : Multiplex mutation screening of the BRCA1 gene in 1000 Japanese breast cancers: 2-3% of Japanese breast cancer is attributable to BRCA1 mutations., Japanese Journal of Cancer Research, Vol.89, No.1, 12-16, 1998.
(要約)
To detect BRCA1 mutations in Japanese breast cancer patients, we screened 1,000 unselected primary cancers for mutations in exon 11, which accounts for 61% of the entire BRCA1 coding sequence. Using a method based on multiplex single-strand conformational polymorphism (SSCP) analysis of multiple restriction fragments generated by restriction-enzyme digestion of amplified DNA, we identified eight mutations. All eight were germline mutations; four of them were non-sense mutations or small deletions resulting in premature stop codons, and the other four were missense mutations. The Japanese carriers of these mutant BRCA1 alleles had developed breast cancers at ages ranging from 45 to 62, five of them bilaterally.
Toyomasa Katagiri, F Kasumi, M Yoshimoto, K Asaishi, R Abe, A Tsuchiya, M Sugano, T Nomizu, S Takai, M Yoneda, K Nanba, M Makita, H Okazaki, K Hirata, M Okazaki, Y Furutsuma, Y Morishita, Y Iino, T Karino, T Fukutomi, H Ayabe, S Hara, T Kajiwara, S Houga, T Shimizu, M Toda, Y Yamasaki, T Uchida, K Kunitomo, H Sonoo, J Kurebayashi, K Shimotuma, Y Nakamura and Y. Miki : High proportion of missense mutations of the BRCA1 and BRCA2 genes in Japanese breast cancer families., Journal of Human Genetics, Vol.43, No.1, 42-48, 1998.
(要約)
Mutations in either of two recently identified genes, BRCA1 and BRCA2, are thought to be responsible for approximately two-thirds of all cases of autosomal-dominantly inherited breast cancer. To examine the nature and frequency of BRCA1 and BRCA2 mutations in Japanese families exhibiting a high incidence of breast cancer, we screened 78 unrelated families in this category for mutations of these two genes. Examining the entire coding sequences as well as exon-intron boundaries of both genes by polymerase chain reaction (PCR) single-strand conformation polymorphism (SSCP) and multiplex-SSCP analysis, we identified possible disease-causing alterations in BRCA1 among affected members of 15 families and in BRCA2 in another 14 families. In 15 of those 29 families, the affected individuals carried missense mutations, although most germline mutations reported worldwide have been deletions or nonsense mutations. Our results, indicating that missense mutations of BRCA1 and BRCA2 tend to predominate over frameshifts or nonsense mutations in Japanese breast cancer families, will contribute significantly to an understanding of mammary tumorigenesis in Japan, and will be of vital importance for future genetic testing.
(キーワード)
BRCA2 Protein / Base Sequence / Breast Neoplasms / DNA, Neoplasm / Female / Genes, BRCA1 / Humans / Japan / Mutation / Neoplasm Proteins / Polymerase Chain Reaction / Polymorphism, Single-Stranded Conformational / Transcription Factors
Toyomasa Katagiri, H Saito, A Shinohara, H Ogawa, N Kamada, Y Nakamura and Y. Miki : Multiple possible sites of BRCA2 interacting with DNA repair protein Rad51., Genes Chromosomes Cancer., Vol.21, 217-22, 1998.
149.
O Maruyama, H Nishimori, Toyomasa Katagiri, Y Miki, A Ueno and Y. Nakamura : Cloning of TCFL5 encoding a novel human basic helix-loop-helix motif protein that is specifically expressed in primary spermatocytes at the pachytene stage, Cytogenetics and Cell Genetics, Vol.82, No.1-2, 41-5, 1998.
(要約)
We have isolated a novel human gene that is expressed specifically in primary spermatocytes in the testis. The cDNA contains an open reading frame of 1356 bp, encoding a 452-amino-acid protein that includes a basic Helix-Loop-Helix (bHLH) motif. The gene, which was mapped to chromosome region 20q13.3-->qter by fluorescence in situ hybridization, consists of six exons and spans approximately 24 kb of genomic DNA. Immunohistochemical staining located the gene product exclusively in cell nuclei of primary spermatocytes at the pachytene stage, but not in those at the leptonema stage. We named this gene TCFL5 (transcription factor-like 5, basic helix-loop-helix). The cell-type and stage-specific expression of TCFL5 indicates that this protein may function in a crucial role in spermatogenesis as a transcription factor by regulating cell proliferation or differentiation of cells through binding to a specific DNA sequence like other bHLH molecules.
Y Miki, Toyomasa Katagiri and Y. Nakamura : Infrequent mutation of the H-Cadherin gene on chromosome 16q24 in human breast cancers., Japanese Journal of Cancer Research, Vol.88, No.8, 701-704, 1997.
(要約)
To investigate the molecular basis of altered expression of the H-cadherin gene, we used polymerase chain reaction-single strand conformation polymorphism and DNA sequencing to examine the H-cadherin gene in 48 primary breast cancers in which loss of the long arm of chromosome 16 had been detected. We identified no mutations other than somatic 5-bp deletion within the coding region in a single tumor. The very low frequency of mutation found in these experiments suggests that H-cadherin is usually not a primary target for carcinogenesis in human breast cancers, and that reduction of its expression is likely to be a consequence of some other genetic event(s).
S Takeda, T Fujiwara, F Shimizu, A Kawai, K Shinomiya, S Okuno, K Ozaki, Toyomasa Katagiri, Y Shimada, M Nagata, T Watanabe, A Takaichi, Y Kuga, M Suzuki, H Hishigaki, E Takahashi, S Shin, Y Nakamura and Y Hirai : Isolation and mapping of karyopherin alpha 3 (KPNA3), a human gene that is highly homologous to genes encoding Xenopus importin, yeast SRP1 and human RCH1., Cytogenetics and Cell Genetics, Vol.76, No.1-2, 87-93, 1997.
(要約)
From a human fetal-brain cDNA library, we isolated and characterized a novel gene (KPNA3) encoding a protein highly homologous to certain nuclear transport proteins of Xenopus and human. The complete cDNA clone, designated karyopherin alpha 3, contained an open reading frame of 1,563 nucleotides encoding 521 amino acids. The predicted amino acid sequence showed 48%, 45% and 48% identity with Xenopus importin, yeast SRP1 and human RCH1, respectively. The similarities among these proteins suggest that karyopherin alpha 3 may be involved in the nuclear transport system. Eight repeats of the arm motif were well conserved among these proteins. The N-terminal region of the predicted karyopherin alpha 3 product was highly basic and the C-terminal region was strongly acidic. A 4.3-kb transcript was expressed in all adult human tissues examined by Northern blotting. The cDNA clone was assigned to chromosome band 13q14.3 by fluorescence in situ hybridization.
F Shimizu, Toyomasa Katagiri, M Suzuki, K T Watanabe, S Okuno, Y Kuga, M Nagata, T Fujiwara, Y Nakamura and E Takahashi : Cloning and chromosome assignment to 1q32 of a human cDNA (RAB7L1) encoding a small GTP-binding protein, a member of the RAS superfamily., Cytogenetics and Cell Genetics, Vol.77, No.3-4, 261-263, 1997.
(要約)
A full-length cDNA homologous to RAB7, a member of the RAB-related GTP-binding protein subfamily, was isolated from a human placenta cDNA library. This cDNA, designated RAB7L1, has an open reading frame of 609 nucleotides encoding 203 amino acids. Northern analysis showed that the mRNA is ubiquitously expressed in human tissues, although signal intensities were different among the various organs examined. This gene was located on chromosome band 1q32 by fluorescence in situ hybridization.
Toyomasa Katagiri, Y Nakamura and Y Miki : Mutations in the BRCA2 gene in hepatocellular carcinomas., Cancer Research, Vol.56, No.20, 4575-4577, 1996.
(要約)
To investigate whether the BRCA2 gene plays a role in carcinogenesis of hepatocellular carcinomas or pancreatic cancers in view of frequent losses of heterozygosity on chromosome 13q12-13 in those tumors, we screened the entire coding region of this gene for mutations in 60 hepatocellular carcinomas and 36 pancreatic cancers. No alteration was found in any of the pancreatic cancers examined, but three mutations were identified in hepatocellular carcinomas; one was a 6-bp somatic deletion within intron 6. The other two mutations we identified in hepatocellular carcinomas were missense mutations in the germ line, although all BRCA2 mutations thus far detected in patients with familial breast cancers likewise have been deletions. None of 194 other patients with cancers or 44 normal controls exhibited either mutation. Combined with our demonstration of BRCA2 expression in adult liver tissue, the evidence implies that inactivation of BRCA2 may play some role in development or progression of hepatocellular carcinoma and might predispose carriers of mutant alleles to liver malignancies.
Y Miki, Toyomasa Katagiri, F Kasumi, T Yoshimoto and Y. Nakamura : Mutation analysis in the BRCA2 gene in primary breast cancers, Nature Genetics, Vol.13, No.2, 245-247, 1996.
(要約)
Breast cancer, one of the most common and deleterious of all diseases affecting women, occurs in hereditary and sporadic forms. Hereditary breast cancers are genetically heterogeneous; susceptibility is variously attributable to germline mutations in the BRCA1 (ref. 1), BRCA2 (ref. 2), TP53 (ref. 3) or ataxia telangiectasia (ATM) genes, each of which is considered to be a tumour suppressor. Recently a number of germline mutations in the BRCA2 gene have been identified in families prone to breast cancer. We screened 100 primary breast cancers from Japanese patients for BRCA2 mutations, using PCR-SSCP. We found two germline mutations and one somatic mutation in our patient group. One of the germline mutations was an insertion of an Alu element into exon 22, which resulted in alternative splicing that skipped exon 22. The presence of a 64-bp polyadenylate tract and evidence for an 8-bp target-site duplication of the inserted DNA implied that the retrotransposal insertion of a transcriptionally active Alu element caused this event. Our results indicate that somatic BRCA2 mutations, like somatic mutations in the BRCA1 gene, are very rare in primary breast cancers.
(キーワード)
BRCA2 Protein / Base Sequence / Breast Neoplasms / DNA Mutational Analysis / DNA Transposable Elements / Female / Germ-Line Mutation / Humans / Molecular Sequence Data / Mutation / Neoplasm Proteins / Repetitive Sequences, Nucleic Acid / Transcription Factors
Toyomasa Katagiri, M Emi, I Ito, K Kobayashi, M Yoshimoto, T Iwase, F Kasumi, Y Miki, H M Skolnick and Y Nakamura : Mutations in the BRCA1 gene in Japanese breast cancer patients., Human Mutation, Vol.7, No.4, 334-339, 1996.
(要約)
Predisposing germline mutations in the BRCA1 gene were identified recently in families with 17 q-linked breast and ovarian cancers. Using single-strand conformation polymorphism (SSCP) analysis, we examined primary breast cancers for mutations in coding exons of BRCA1 in a panel of 103 patients, of whom all either represented early-onset cases (< 35 of age), were members of multiply-affected families, and/or had developed bilateral breast cancers. Mutations were detected in tumors from four patients, all of whom had developed breast cancers bilaterally: a frame-shift due to a 2-bp deletion at codon 797; a nonsense mutation at codon 1214; and two missense mutations, one at codon 271 leading to Val-->Met substitution, and the other at codon 1150 leading to Pro-->Ser substitution. In each case the same mutation was present in constitutional DNA. The mean age of onset was 49 years among the Japanese carriers of BRCA1 mutations identified in this study, in contrast to the mean age of 35 observed among carriers of BRCA1 mutations in a similar U.S. study (Futreal et al., 1994). The evidence reported here supports a rather limited role of BRCA1 in breast carcinogenesis.
(キーワード)
Adult / BRCA1 Protein / Base Sequence / Breast Neoplasms / DNA Primers / DNA, Neoplasm / Genetic Carrier Screening / Humans / Japan / Middle Aged / Molecular Sequence Data / Mutation / Neoplasm Proteins / Polymorphism, Single-Stranded Conformational / Transcription Factors
Toyomasa Katagiri, K Ozaki, T Fujiwara, F Shimizu, A Kawai, S Okuno, M Suzuki, Y Nakamura, E Takahashi and Y. Hirai : Cloning, expression, and chromosome mapping of adducin-(alpha) like 1 (ADDL1), a human cDNA highly homologous to human erythrocyte adducin, Cytogenet Cell Genet., Vol.74, 90-5, 1996.
157.
TK* Watanabe, Toyomasa Katagiri, M Suzuki, F Shimizu, T Fujiwara, N Kanemoto, Y Nakamura, Y Hirai, H Maekawa and E. Takahashi : Cloning and characterization of two novel human cDNAs (NELL1 and NELL2) encoding proteins with six EGF-like repeats, Genomics, Vol.38, 273-6, 1996.
158.
I Ito, M Yoshimoto, T Iwase, S Watanabe, Toyomasa Katagiri, Y Harada, F Kasumi, S Yasuda, T Mitomi and M Emi : Association of genetic alterations on chromosome 17 and loss of hormone receptors in breast cancer., British Journal of Cancer, Vol.71, No.3, 438-441, 1995.
(要約)
To investigate possible relationships between genetic alterations and hormonal deregulation during breast cancer development and/or progression, we examined 616 primary breast cancers for loss of heterozygosity (LOH) at chromosomal regions 16q24, 17p13.3 and 17q21, and for amplifications of the ERBB2 and c-MYC loci. A comparison of oestrogen receptor (ER) and progesterone receptor (PgR) status in tumour cells with data concerning these genetic alterations revealed that LOH at 17q21 was significantly correlated with absence of oestrogen receptors (ER) (P < 0.0003) or progesterone receptors (PgR) (P < 0.0001), and with the absence of both (P < 0.0001). Similarly, a significant association was observed between amplification of ERBB2 and the absence of either ER or PgR. LOH at 17p13.3 was associated with the absence of PgR (P < 0.01). These data suggest a possible relationship between specific genetic changes on chromosome 17 and hormonal deregulation in the progression of breast cancer.
Toyomasa Katagiri, Y Harada, M Emi and Y Nakamura : Human metalloprotease/disintegrin-like (MDC) gene: exon-intron organization and alternative splicing., Cytogenetics and Cell Genetics, Vol.68, No.1-2, 39-44, 1995.
(要約)
A recently identified gene encoding a metalloprotease-like, disintegrin-like, cysteine-rich protein (MDC) represents a candidate tumor suppressor gene for human breast cancer based on its location within a minimal region of chromosome 17q21 previously defined by tumor deletion mapping. The work reported here has shown that the MDC gene consists of 28 exons interrupted by relatively short introns, most of them 67 bp to 5 kb in length. We have identified two forms of transcripts generated by alternative splicing. The more abundant form encodes a protein of 769 amino acids; the other, a previously described cDNA, encodes 524 amino acids. Exons 1a, 1b, 1c, 1d, and 2-7 encode a proprotein domain; exons 7-13, a metalloprotease-like domain; exons 14-17, a disintegrin domain; exons 18-22, a cysteine-rich domain, including an epidermal growth factor (EGF)-like repeat domain within exons 21 and 22; exon 23, a transmembrane domain; and exons 24 and 25, a short cytoplasmic domain. These results show that human MDC contains a mosaic of exons capable of encoding several functional domains.
M Ichii, Toyomasa Katagiri, H Hasegawa, O Yatou, DW Kirk and JL. Wray : Biochemical characterization of rice (Oryza sativa L.) mutants defective or low in nitrate reduction, Tech Bull Fac Agric Kagawa University, Vol.47, 1-6, 1995.
161.
M Isomura, A Tanigami, H Saito, Y Harada, Toyomasa Katagiri, J Inazawa, H D Ledbetter and Y Nakamura : Detailed analysis of loss of heterozygosity on chromosome band 17p13 in breast carcinoma on the basis of a high-resolution physical map with 29 markers., Genes, Chromosomes & Cancer, Vol.9, No.3, 173-179, 1994.
(要約)
We have constructed a physical map of chromosome band 17p13, using 29 markers that had been localized to 17p13 by means of fluorescence in situ hybridization (FISH) and analysis by pulsed-field gel electrophoresis (PFGE). The map spans nearly 8 Mb of genomic DNA, and the estimated average distance between each marker is roughly 290 kb. The p13 band of chromosome 17 is thought to contain a putative tumor suppressor gene in addition and distal to TP53. Deletion mapping in a large number of breast carcinomas indicated that the tumor suppressor gene lies between the loci defined by cC117-708 (D17S878) and p144D6 (D17S34), which are an estimated 7 Mb apart. Our results should contribute to construction of a contig map of chromosome band 17p13 with cosmid and/or YAC (yeast artificial chromosome) clones, and to isolation of the putative tumor suppressor gene.
M Isomura, A Tanigami, H Saito, Y Harada, Toyomasa Katagiri, J Inazawa, DH Ledbetter and Y. Nakamura : Detailed analysis of heterozygosity on chromosome band 17p13 in breast carcinoma on the basis of a high-resolution physical map with 29 markers, Genes Chromosomes Cancer, Vol.9, 173-9, 1994.
163.
Y; Harada, Toyomasa Katagiri, I Ito, F Akiyama, G Sakamoto, F Kasumi, Y Nakamura and M. Emi : Genetic studies of 457 breast cancers. Clinicopathologic parameters compared with genetic alterations., Cancer., Vol.74, 2281-6, 1994.
164.
M Emi, Toyomasa Katagiri, Y Harada, H Saito, J Inazawa, I Ito, F Kasumi and Y. Nakamura : A novel metalloprotease/ disintegrin-like gene at 17q21.3 is somatically rearranged in two primary breast cancers, Nature Genetics, Vol.5, No.2, 151-157, 1993.
(要約)
From chromosomal region 17q21.3, where a tumour suppressor gene(s) for breast and ovarian cancers is thought to be present, we have isolated a novel gene from a cosmid clone that revealed somatic rearrangements in two breast cancers. The gene (MDC) encodes a 524-amino acid metalloprotease-like, disintegrin-like and cysteine-rich protein with sequence similarity to members of the snake-venom metalloprotease/disintegrin family and guinea-pig sperm-surface protein PH-30. These proteins have been implicated in cell-cell or cell-extracellular matrix interactions. Rearrangements in both tumours involve multiple exons and disrupt the coding region of the new MDC.
M Ichii, Toyomasa Katagiri and H Hasegawa : Mutants with low nitrate reductase activity selected from seedlings expressing nitrogen deficiency symptoms in rice (Oryza sativa L.), Jpn J Breed., Vol.43, 123-7, 1993.
166.
H Hasegawa, Toyomasa Katagiri, S Ida, O Yatou and M. Ichii : Characterization of a rice (Oryza sativa L.) mutant deficient in the heme domain of nitrate reductase, Theor Appl Genet., Vol.84, 6-9, 1992.
167.
H Hasegawa, O Yatou, Toyomasa Katagiri and M. Ichii : Screening for nitrate reductase deficient mutants in rice (Oryza sativa L.), Jpn J Breed., Vol.41, 95-101, 1991.
学術論文(紀要・その他):
1.
Daishiroh Kobayashi, Masaya Denda, JUNYA Hayashi, Kohta Hidaka, Yutaka Kohmura, Takaaki Tsunematsu, Kohei Nishino, Harunori Yoshikawa, OHKAWACHI Kento, Kiyomi Nigorikawa, Tetsuro Yoshimaru, Naozumi Ishimaru, Nomura Wataru, Toyomasa Katagiri, Hidetaka Kosako and Akira Otaka : Sulfoxide-mediated Cys-Trp-selective bioconjugation that enables protein labeling and peptide heterodimerization, ChemRxiv, 2024.
C Fukukawa, Yusuke Nakamura and Toyomasa Katagiri : Molecular target therapy for synovial sarcoma., Future Oncology, Vol.1, No.6, 805-812, Dec. 2005.
(要約)
Although the specific chromosomal translocation and fusion gene SYT-SSX in synovial sarcoma (SS) has been identified, the molecular mechanism of its tumorigenesis is largely unknown. Recent gene-expression profiles of soft-tissue tumors using cDNA microarray demonstrated that SS has the distinct gene-expression pattern from other sarcomas and has a similar pattern to that of malignant peripheral nerve sheath tumors, indicating that the origin of SS is likely to be the neural crest cells. Through this analysis, several genes were found to be specifically upregulated in SS and considered to play an important role in the proliferation of SS cells. Among them, Frizzled homolog 10 was identified as a good candidate molecule for the development of novel therapies to treat SS patients.
Chiho Shinozaki, Yutaka Kohmura, Tetsuro Yoshimaru, Tsuyoshi Tahara, Masaya Denda, Hidefumi Mukai, Kohta Mohri, Yi Long Chen, Toyomasa Katagiri and Akira Otaka : Study on a lipidated anti-cancer peptide allowing long-lasting duration in mice model, AIMECS 2023, Seoul, Jun. 2023.
2.
Takeshi Harada, Asuka Oda, Yosuke Matsushita, Ryohei Sumitani, Yusuke Inoue, Tomoyo Hara, Masahiro Oura, Kimiko Sogabe, Tomoko Maruhashi, Mamiko Takahashi, Kiyoe Kurahashi, Shiroh Fujii, Shingen Nakamura, Hirokazu Miki, Masahiro Hiasa, Jumpei Teramachi, Toyomasa Katagiri and Masahiro Abe : ADAR1-dsRNA metabolism in myeloma cells with 1q amplification: a novel therapeutic target, 19th International Myeloma Society Annual Meeting, Aug. 2022.
3.
Toyomasa Katagiri and Yosuke Matsushita : Genetic and epigenetic alterations of SALL3 contributes to chemoresistance in triple-negative breast cancer, the 5th Annual Meeting of the International Society of Precision Cancer Medicine (ISPCM), virtual meeting, Online, Sep. 2021.
4.
Tetsuro Yoshimaru, Yosuke Matsushita, Sasa Mitsunori, Miyoshi Yasuo and Toyomasa Katagiri : BIG3 phosphatase inactivates tumor suppressor PHB2 via tis dephosphorylation to contribute to the breast carcinogenesis, The 14th International Conference on Protein Phosphatase, Online, Dec. 2020.
5.
Toyomasa Katagiri : Targeting BIG3-PHB2 interaction to overcome endocrine-resistant breast cancer, Satellite Meeting of International Society of Precision Cancer Medicine in Korea, Yeosu, Nov. 2019.
6.
Yosuke Matsushita, Masato Komatsu, Kazuma Kiyotani, Tetsuro Yoshimaru, Suzuki Hiromu, Miyoshi Yasuo, Sasa Mitsunori and Toyomasa Katagiri : Frequent downregulation of SALL3 by genetic and epigenetic alterations is involved in progression and chemoresistance of triple negative breast cancers, American Association for Cancer Research ANNUAL MEETING 2019, Atlanta, Apr. 2019.
7.
Tetsuro Yoshimaru, Yosuke Matsushita, Sasa Mitsunori, Miyoshi Yasuo and Toyomasa Katagiri : PHB2 inactivation by AKAP-BIG3 is required for progression of HER2-overexpressing breast cancer, American Association for Cancer Research ANNUAL MEETING 2019, Atlanta, Apr. 2019.
8.
Toyomasa Katagiri : Targeting RHBDL2-SLC1A5 axis to overcome chemoresistance and progression intriple negative breast cancer, International Society of Precision Cancer Medicine (ISPCM) Annual Meeting 2019, Seoul, Mar. 2019.
9.
Keitaroh Anyohji, Keisuke Aihara, Tetsuro Yoshimaru, Akira Shigenaga, Toyomasa Katagiri and Akira Otaka : Development of anti-cancer peptide based on prohibitin 2, 10th International Peptide Symposium, Kyoto, Dec. 2018.
10.
Toyomasa Katagiri : Novel therapeutic strategy for breast cancer utilizing activation of tumor suppressor PHB2, The 2nd International Symposium on Radiation Therapeutics and Biology The 34th Radiation Biology Center Internationl Symposium, Kyoto, Nov. 2018.
11.
Toyomasa Katagiri : Comprehensive molecular features of triple negative breast cancers, The 13th International Symposium of the Institute Network for Biomedical Sciences joint with the 3rd Symposium of the Inter-University Research Network for Trans-Omics Medicine and the 28th Hot Spring Harbor Symposium, Fukuoka, Oct. 2018.
12.
Yosuke Matsushita, Masato Komatsu, Kazuma Kiyotani, Tetsuro Yoshimaru, Suzuki Hiromu, Miyoshi Yasuo, Sasa Mitsunori and Toyomasa Katagiri : Frequent downregulation of SALL3 by recurrent genetic and epigenetic alterations is involved in triple-negative breast cancers, American Association for Cancer Research ANNUAL MEETING 2018, Vol.78, No.13, Chicago, Apr. 2018.
Tetsuro Yoshimaru, Yosuke Matsushita, Sasa Mitsunori, Miyoshi Yasuo and Toyomasa Katagiri : Overcoming trastuzumab resistance in HER2-overexpressing breast cancer by utilizing PHB2, a tumor suppressor of multiple resistance pathways, American Association for Cancer Research ANNUAL MEETING 2018, Chicago, Apr. 2018.
14.
Toyomasa Katagiri, Kei Daizumoto, Tetsuro Yoshimaru, Yosuke Matsushita, Tomoya Fukawa, Ono Masaya and Hiro-omi Kanayama : DDX31 cooperates with mutant p53 and EGFR to promote the multistep progression of invasive bladder cancer, American Association For Cancer Research ANNUAL MEETING 2018, Chicago, Apr. 2018.
15.
Toyomasa Katagiri : Stapled BIG3 helical peptide ERAP potentiates anti-tumor activity for breast cancer therapeutics, International Society of Precision Cancer Medicine Annual Meeting 2018, Busan, Mar. 2018.
16.
Akira Otaka, Keisuke Aihara, Tetsuro Yoshimaru, Akira Shigenaga and Toyomasa Katagiri : Development of long-lasting stapled peptides targeting BIG3-PHB2 interaction in breast cancer cells, The 6th Pharmaceutical Sciences World Congress 2017, Stockholm, May 2017.
17.
Toyomasa Katagiri : Development of chemically modified peptide inhibitor ERAP targeting BIG3-PHB2 complex on hormone-resistant breast tumor, American Association For Cancer Research ANNUAL MEETING 2017, Washington, D.C., Apr. 2017.
18.
Tetsuro Yoshimaru and Toyomasa Katagiri : Development of chemically modified peptide inhibitor ERAP targeting BIG3-PHB2 complex on hormone-resistant breast cancer, 2nd International Symposium of Molecular Medicine in Tokushima University, Tokushima, Nov. 2016.
19.
Tetsuro Yoshimaru, Ono Masaya, Mizuguchi Kenji, Miyoshi Yasuo, Mitsunori Sasa and Toyomasa Katagiri : A novel A-kinase anchoring protein BIG3, coordinates estrogen signalling in breast cancer cells, The 12th International Conference on Protein Phosphatase, Oct. 2016.
20.
Toyomasa Katagiri : Novel targeting therapeutic strategy for treatment of endocrine resistant breast cancer, Tne 34th Sapporo International Cancer Symposlum, Sapporo, Jun. 2015.
21.
Toyomasa Katagiri : Xanthohumol suppresses estrogen-signaling in endocrine resistant breast cancer through the specific inhibition of BIG3-PHB2 interactions, American Association for Cancer Research (AACR) Annual Meeting 2015, Pennsylvania, Apr. 2015.
22.
Toyomasa Katagiri : Targeting BIG3-PHB2 interaction to overcome endocrine resistance in breast cancer cells, 2015 SNUCRI & SNUCH CANCER SYMPOSIUM, Hwasun, Korea, Apr. 2015.
23.
Toyomasa Katagiri : A novel AKAP protein, BIG3 coodinates estrogen signaling pathways in breast cancer cells., 11th International Conference on Protein Phosphatase, Sendai, Nov. 2014.
24.
Toyomasa Katagiri, Tetsuro Yoshimaru, Masato Komatsu and Taisuke Matsuo : BIG3-PHB2 Interaction is a key therapeutic target in Iuminal-type of breast cancer., Amerian Association for Cancer Research (AACR) Annal Meeting 2014, San Diego, Apr. 2014.
25.
Toyomasa Katagiri : Overcoming Endocrine Resistance in Breast Cancer by Reactivation of Tumor Suppressor Protein, 2013 SNUCRI & SNUCH CANCER SYMPOSIUM, JEJU, May 2013.
26.
Toyomasa Katagiri, Tomoya Fukawa, Taisuke Matsuo and Hiro-omi Kanayama : DDX31 regulates p53 tumor suppressive activity in renal cell carcinomas, Amerian Association for Cancer Research (AACR) Annal Meeting 2013, Washington, D.C., Apr. 2013.
27.
Toyomasa Katagiri : Regulation of estrogen-signaling in ER-positive breast cancer by tumor suppressor REA via ERAP1, 15th International Congress on Hormonal Steroids and Hormones & Cancer, Kanazawa, Nov. 2012.
28.
Taisuke Matsuo, Ono Masaya, Tetsuro Yoshimaru, Miyoshi Yasuo, Sasa Mitsunori, Nakamura Yusuke and Toyomasa Katagiri : Functional roles of a novel glycosyltrasnferase BCGT1 in breast cancer cells, 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
29.
Toyomasa Katagiri : Novel targeting therapeutic strategy for treatment of estrogen-dependent breast cancer, 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
30.
Furu Moritoshi, Kajita Yoichiro, Nagayama Satoshi, Toyomasa Katagiri, Nakamura Yusuke and Toguchida Junya : Identifiication of AFAP1L1 as a prognostic marker for spindle cell sarcomas, 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
31.
Elgazzar Seham, Zembutsu Hitoshi, Takahashi Atsushi, Nakashima Mitsuko, Kubo Michiaki, Toyomasa Katagiri, Miki Yoshio, Kamatani Naoyuki and Nakamura Yusuke : A genome-wide association study identifies a locus associated with breast cancer in Japanese., Oct. 2011.
32.
Yanai Ayako, Ito Takashi, Hirota Seiichi, Toyomasa Katagiri, Sasa Mitsunori, Tomita Naohiro and Miyoshi Yasuo : Estrogen receptors may be down-regulated in pre-,and rare in postmemopausal breast cancers with low proliferation, 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
33.
Tomoya Fukawa, Ono Masaya, Hisanori Uehara, Taisuke Matsuo, Nakamura Yusuke, Hiro-omi Kanayama and Toyomasa Katagiri : Identification and characterization of RCCDH as a novel molecular target for clear renal cell carcinoma., 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
34.
Tetsuro Yoshimaru, Komatsu Masato, Taisuke Matsuo, Miyoshi Yasuo, Sasa Mitsunori, Nakamura Yusuke and Toyomasa Katagiri : Novel modeling of ER signaling regulation in E2-dependent breast cancer - New therapeutic target for ERAP1-REA complex, 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
Park Jae-Hyun, Fukukawa Chikako, Toyomasa Katagiri and Nakamura Yusuke : Identification of an O-type glycosyltransferase(GALNT6)as a nobel molecular target for breast cancer therapy, 69th Annual Meeting of the Japanese Cancer Association, Sep. 2010.
Fukukawa Chikako, Nishidate Toshihiko, Nakamura Yusuke and Toyomasa Katagiri : Analysis of cell-growth promoting role of BGPRP in breast cancer, 69th Annual Meeting of the Japanese Cancer Assosiation, Sep. 2010.
Toyomasa Katagiri, Tetsuro Yoshimaru, Taisuke Matsuo, Miyoshi Yasuo, Sasa Mitsunori and Nakamura Yusuke : Critical role of transactivation of ERAP1,estrogen receptor activity-regulated protein 1, in estrogen-dependent breast cancer cell growth, The 6th International Symposium on Hormonal Carcinogenesisi in Japan, Sep. 2010.
42.
Wataru Obara, Mitsugu Kanehira, Ryo Takata, Takuya Tsunoda, Koji Yoshida, Toyomasa Katagiri, Yusuke Nakamura and Tomoaki Fujioka : Phase I clinical trial of novel HLA-A24 restricted DEPDC1 and MPHOSPH1 peptide vaccine for bladder cancer., 8th Joint Conference of the American Association for Cancer Research and the Japanese Cancer Association, Waikoloa, Feb. 2010.
43.
Harada Yosuke, Toyomasa Katagiri, Kanehira Mitsugu, Takata Ryo, Fujioka Tomoaki and Nakamura Yusuke : Investigation of a novel potential therapeutic modality targeting to DEPDC1 for bladder cancer., 8th Joint Conference of the American Association for Cancer Research and the Japanese Cancer Association, Waikoloa, Feb. 2010.
44.
Ajiro Masahiko, Toyomasa Katagiri, Ueda Koji, Nakagawa Hidewaki, Nishidate Toshihiko, Fukukawa Chikako, Daigo Yataro and nakamura Yusuke : Involvement of RQCD1 overexpression, a novel cancer-testis antigen, in breast cancer carcinogenesis though the regulation fo Akt activity, 8th Joint Conference of the American Association for Cancer Research and the Japanese Cancer Association, Waikoloa, Feb. 2010.
45.
Toyomasa Katagiri and Yusuke Nakamura : Critical role of transactivation of ERAP1, estrogen receptor activity-regulated protein 1, in estrogen-dependent breast cancer cells., 8th Joint Conference of the American Association for Cancer Research and the Japanese Cancer Association, Waikoloa, Feb. 2010.
46.
Nagayama Satoshi, Takahashi Ryo, Imoto Seiya, Furu Moritoshi, Toyomasa Katagiri, Nakamura Yusuke, Toguchida Junya and Sakai Yoshiharu : Functional and immunohistochemical analyses of a novel protein upregulated in colorectal cancers., 8th Joint Conference of the American Association for Cancer Research and the Japanese Cancer Association, Waikoloa, Feb. 2010.
47.
Kim Jung-Won, Toyomasa Katagiri, Fukukawa Chikako, Daigo Yataro, Nishidate Toshihiko and Nakamura Yusuke : Identification and characterization of BCUP2 as a novel molecular target for breast cancer therapy., 8th Joint Conference of the American Association for Cancer Research and the Japanese Cancer Association, Waikoloa, Feb. 2010.
48.
Fukukawa Chikako, Toyomasa Katagiri, Hanaoka Hirofumi, Nagayama Satoshi, Toguchida Junya, Yoshioka Hiroki, Endo Keigo and Nakamura Yusuke : Development of antibody therapy against synovial sarcoma-targeting novel antigenidentified through analysis of genome-wide gene expression profile, 8th Joint Conference of the American Association for Cancer Research and the Japanese Cancer Association, Waikoloa, Feb. 2010.
49.
Toyomasa Katagiri and Nakamura Yusuke : Strategy for development of novel molecular-targeting drugs for breast cancer through gene-expression profile analysis, Workshop Mexico-Japan "Genomic Medicine", Tokyo, Nov. 2009.
50.
Nagayama Satioshi, Furu Moritoshi, Imoto Seiya, Takahashi Ryo, Kubo Hajime, Toyomasa Katagiri, Nakamura Yusuke, Toguchida Junya and Sakai Yoshiharu : A novel recurrence predictor upregulated in colorectal cancers, 68th Annual Meeting of the Japanese Cancer Association, Yokohama, Oct. 2009.
51.
Toyomasa Katagiri and Nakamura Yusuke : Strategy for development of novel molecular-targeting drugs for breast cancer through gene-expression profile analysis, 68th Annual Meeting of the Japanese Cancer Association, Yokohama, Oct. 2009.
52.
Park Jae-Hyun, Fukukawa Chikako, Nakamura Yusuke and Toyomasa Katagiri : Identification and characterization of a novel therapeutic target with glycosylation activity in mammary carcinogenesis, 68th Annual Meeting of the Japanese cancer Association, Yokohama, Oct. 2009.
53.
Harada Yosuke, Toyomasa Katagiri, Kanehira Mitsugu, Takata Ryo, Fujioka Tomoaki and Nakamura Yusuke : Investigation of a novel potential therapeutic modality targeting to DEPDC1 for bladder cancer, 68th Annual Meeting of the Japanese cancer Association, Yokohama, Oct. 2009.
54.
Ajiro Masahiko, Toyomasa Katagiri, Nishidate Toshihiko, Fukukawa Chikako, Daigo Yatoro and Nakamura Yusuke : Characterazation of a novel therapeutic molecule, BCUP1 involved in mammary carcinogenesis, 68th Annual Meeting of the Japanese Cancer Association, Yokohama, Oct. 2009.
55.
Kim Jung-Won, Toyomasa Katagiri, Fukukawa Chikako, Daigo Yatoro, Nishidate Toshihiko and Nakamura Yusuke : Identification and characterization of BCUP2 as a novel molecular target for breast cancer therapy, 68th Annual Meeting of the Japanese Cancer Association, Yokohama, Oct. 2009.
56.
Toyomasa Katagiri : Identification of PBK/TOPK, Ser/Thr protein kinase as a druggable-target though analysis of genome-wide gene expression profile, The 7th Sino-Japan Joint Conference for Cancer Research, Dec. 2008.
57.
片桐 豊雅, 中村 祐輔 :
(International symposium, oral). Second JCA-AACR Special Joint Conference. The Latest Advances in Breast Cancer Research: From Basic Science to Therapeutics. Identification of PBK/TOPK, Ser/Thr protein kinase, as a druggable-target for breast cancer therapy though analysis of genome-wide gene expression profile. (International symposium, oral), Awaji-island, 2008年7月.
58.
C Fukukawa, Toyomasa Katagiri, H Hanaoka, S Nagayama, J Toguchida, K Endo and Y Nakamura : Radioimmunotherapy for human synovial sarcoma xenografts with Yttrium 90-labeled anti-FZD10 monoclonal antibody, Proceedings of the 99th Annual Meeting of the American Association for Cancer Research, San Diego, CA. Philadelphia (PA), Apr. 2008.
59.
S Nagayama, M Furu, T Aoyama, H Kubo, G Watanabe, Toyomasa Katagiri and Y Nakamura : Toguchida J, Sakai Y. Immunohistochemical analysis of a novel protein associated with the development of colorectal cancers, Proceedings of the 99th Annual Meeting of the American Association for Cancer Research, San Diego, CA. Philadelphia (PA), Apr. 2008.
60.
H Zembutsu, Y Suzuki, A Sasaki, T Tsunoda, T Hasegawa, DN Nikolova, LS Kee, Toyomasa Katagiri, K Hirata and Y Nakamura : Predicting response to docetaxel neoadjuvant chemotherapy for advanced breast cancers through genome-wide gene expression profiling, Proceedings of the 99th Annual Meeting of the American Association for Cancer Research, San Diego, CA. Philadelphia (PA), Apr. 2008.
61.
K Ueda, Y Daigo, Y Fukase, Toyomasa Katagiri, N Ishikawa, S Irie, T Sato, N Kohno, M Shiwa and Y Nakamura : A novel glycoproteomic approach for the discovery of carbohydrate-targeting serum tumor markers for lung cancer using Lectin-coupled ProteinChip system, Proceedings of the 99th Annual Meeting of the American Association for Cancer Research, San Diego, CA. Philadelphia (PA), Apr. 2008.
62.
J Park, Toyomasa Katagiri and Y Nakamura : PBK/TOPK, a mitotic Ser/Thr kinase, is a novel druggable target for breast cancer therapy, Proceedings of the 99th Annual Meeting of the American Association for Cancer Research, San Diego, CA. Philadelphia (PA), Apr. 2008.
63.
Toyomasa Katagiri, M Lin, J Park and Y Nakamura : Identification of MELK, Ser/Thr protein kinase, as a druggable-target for breast cancer therapy though analysis of genome-wide gene expression profile, Proceedings of the 99th Annual Meeting of the American Association for Cancer Research, San Diego, CA. Philadelphia (PA), Apr. 2008.
64.
S Dobashi, Toyomasa Katagiri, E Hirota, T Shuin, T Fujioka, T Miki and Y Nakamura : Identification and characterization of RAPTM as a novel therapeutic target for renal cell carcinoma, Proceedings of the 99th Annual Meeting of the American Association for Cancer Research, San Diego, CA. Philadelphia (PA), Apr. 2008.
65.
J Kim, M Akiyama, Toyomasa Katagiri, T Nishidate and Y Nakamura : Identification and characterization of ERAP1 as a novel molecular target for breast cancer therapy, Proceedings of the 99th Annual Meeting of the American Association for Cancer Research, San Diego, CA. Philadelphia (PA), Apr. 2008.
66.
Toyomasa Katagiri and Nakamura Yusuke : Development of prediction system of response to M-VAC neoadjuvant chemotherapy for bladder cancer toward personalized medicine, The 2007 EAUHGS symposium & the 7th EAUHGS Annual meeting, Changsha, Funan, China, Dec. 2007.
67.
Toyomasa Katagiri and Nakamura Yusuke : Development of prediction system of response to M-VAC neoadjuvant chemotherapy for bladder cancer, 7th Joint conference of the American association for cancer research and the Japanese cancer association, Jan. 2007.
Yosuke Matsushita, Kazumasa Okumura, Masato Komatsu, Tetsuro Yoshimaru, Ono Masaya, Akira Tangoku, Miyoshi Yasuo, Sasa Mitsunori and Toyomasa Katagiri : RHBDL2 has essential roles for glutaminolysis and chemoresistance in triple negative breast cancer, The 17th International Symposium of the Institute Network for Biomedical Sciences International Symposium on Tumor Biology in Kanazawa 2022, Oct. 2022.
13.
Tetsuro Yoshimaru, Yosuke Matsushita, Hidetaka Kosako, Sasa Mitsunorii, Miyoshi Yasuo and Toyomasa Katagiri : The plasma membrane BIG3-PHB2 complex contributes to the acquisition of trastuzumab-resistance in HER2-positive breast cancer, The 17th International Symposium of the Institute Network for Biomedical Sciences International Symposium on Tumor Biology in Kanazawa 2022, Oct. 2022.
Abdullah S. Ili, Tetsuro Yoshimaru, Yosuke Matsushita, Masato Komatsu, Miyoshi Yasuo, Honda Junko, Ohsumi Shozo, Sasa Mitsunori and Toyomasa Katagiri : Whole-exome sequencing for the identification of Japanese familial breast cancer susceptibility genes, The 81st Annual Meeting of the Japanese Cancer Association, Oct. 2022.
Abdullah S. Ili, Yosuke Matsushita, Yasuko Takahashi, Masato Komatsu, Kazuma Kiyotani, Yasuo Miyoshi, Junko Honda, Shozo Ohsumi, Mitsunori Sasa and Toyomasa Katagiri : Identification and characterization of novel susceptibility genes in hereditary Japanese familial breast cancer, 第80回日本癌学会学術総会, Sep. 2021.
Boya Deng, Yunus Tarhan Emre, Lili Ren, Ueda Koji, Toyomasa Katagiri, Jae-Hyun Park and Nakamura Yusuke : Critical role of O-glycosylation of estrogen receptor alpha by GALNT6 in breast cancer cells, The 77th Annual Meeting of the Japanese Cancer Association, Sep. 2018.
82.
K.Li Huizi, Kanda Hiroaki, Nagayama Satoshi, Toyomasa Katagiri, Nakamura Yusuke and Hasegawa Sumitaka : A novel therapeutic option for synovial sarcoma using alpha-radiolabeled FZD10 antibody, The 77th Annual Meeting of the Japanese Cancer Association, Sep. 2018.
Toyomasa Katagiri : Regulation of estrogen/ estrogen receptor signallings in breast cancer cells, 11th International Symposium of The Institute Network ``Frontiers in Biomedical Sciences'', Jan. 2017.
103.
粟飯原 圭佑, 吉丸 哲郎, 小松 正人, 重永 章, 片桐 豊雅, 大髙 章 : Development of stapled peptides targeting BIG3-PHB2 interaction in breast cancer cells, 第53回ペプチド討論会, 2016年10月.
Jiaying Lin, Jae-Hyun Park, Suyoun Chung, Koji Ueda, Toyomasa Katagiri, Koichi Matsuda and Yusuke Nakamura : GALNT6 stabillizes GRP78 protein by O-type glycosylation and enhances its activity to suppress apoptosis under stress, The 73rd Annual Meeting of the Japanese Cancer Association, Sep. 2014.
Nakamura Toru, Toyomasa Katagiri, Nakagawa Hidewaki, Tsuchikawa Takahiro, Hirano Satoshi and Nakamura Yusuke : Isolation and characterization of a novel gene as a therapeutic target for pancreatic cancer, 72nd Annual Meeting of the Japanese Cancer Association72, Oct. 2013.
141.
Lo Hau Yi Paulisally, tanikawa Chizu, Toyomasa Katagiri, Nakamura Yusuke and Matsuda Koichi : Identification of candidate tumor suppressor gene LBC1 in breast cancer, 72nd Annual Meeting of the Japanese Cancer Association, Oct. 2013.
Toyomasa Katagiri : New insight into therapeutic strategies for acquired endocrine resistant breast cancer, 72nd Annual Meeting of the Japanese Cancer Association, Oct. 2013.
Nagayama Satoshi, Takahashi Ryo, Imoto Seiya, Furu Moritoshi, Kajita Yoichiro, Toyomasa Katagiri, Nakamura Yusuke, Sakai Yoshiharu and Toguchida Junya : Cytoskeleton-related protein C7059 is involved in the progression of colorectal cancer via regulation of cell migration, 72nd Annual Meeting of the Japanese Cancer Association, Oct. 2013.
147.
Siew-Kee Low, Takahashi Atsushi, Kubo Michiaki, Toyomasa Katagiri and Nakamura Yusuke : Genome-wide association study and pathway analysis for the genetic risk of breast cancer in japanese population, 72nd Annual Meeting of the Japanese Cancer Association, Oct. 2013.
148.
Masato Komatsu, Tetsuro Yoshimaru, Taisuke Matsuo, Kazuma Kiyotani, Miyoshi Yasuo, Sasa Mitsunori and Toyomasa Katagiri : Nuclear-19S proteasome associated gene 1 contributes to the aggressiveness of triple negative breast cancer cells, 72nd Annual Meeting of the Japanese Cancer Association, Oct. 2013.
ELGAZZAR Seham, Takahashi Norihiko, Yamaguchi Kiyoshi, Zembutsu Hitoshi, Toyomasa Katagiri, Ikenoue Tsuneo, Nakamura Yusuke and Furukawa Yoichi : Identification of a single nucleotide polymorphism associated with SIAH2 Expression, 72nd Annual Meeting of the Japanese Cancer Association, Oct. 2013.
Siew-Kee Low, Takahashi Atsushi, Seham Elgazzar, Kugo Michiaki, Toyomasa Katagiri and nakamura Yusuke : Genome-wide association study of breast cancer in Japanese population, The 71th Annual Meeting of the Japanese Cancer Association, Sep. 2012.
ELGAZZAR SEHAM, Zembutsu Hitoshi, Takahashi Atsushi, Kubo Michiaki, Aki Fuminori, Hirata Koichi, Takatsuka Yuichi, Okazaki Minoru, Ohsumi Shozo, Miki Yoshio, Sasa Mitsunori, Toyomasa Katagiri and Nakamura Yusuke : A genome-wide association study identifies a locus associated with hormonal receptor positive breast cancer in Japanese, The 71th Annual Meeting of the Japanese Cancer Association, Sep. 2012.
Deng Zhenzhong, Lin Jiaying, Tanikawa Chizu, Lo Hau Yi Paulisally, Toyomasa Katagiri, Daigo Yataro, Furukawa Hoichi, Nakamura Yusuke and Matsuda Koichi : Screenting of expression profiles consisting of 783 cancer tissues identified TSRCC2 as a putative tumor suppressor gene, The 71th Annual Meeting of the Japanese Cancer Association, Sep. 2012.
166.
Lin Jiaying, Deng Zhenzhong, Lo Hau Yi Paulisally, Tanikawa Chizu, Toyomasa Katagiri, Nakamura Yusuke and Matsuda Koichi : Genome-wide gene expression analyses identified TSRCC1 as a potential tumor suppressor gene of renall cell carcinoma, The 71th Annual Meeting of the Japanese Cancer Association, Sep. 2012.
Le T. Dat, Taisuke Matsuo, Tetsuro Yoshimaru, Souji Kakiuchi, Hisatsugu Goto, Kuramoto Takuya, Mitsuhashi Atsushi, Saburo Sone, Yasuhiko Nishioka and Toyomasa Katagiri : LBMTF a novel transcriptional factor involved in lung cancer bone metastases, The 71th Annual Meeting of the Japanese Cancer Association, Sep. 2012.
169.
小松 正人, 吉丸 哲郎, 松尾 泰佑, 清谷 一馬, 三好 康雄, 島田 光生, 中村 祐輔, 笹 三徳, 片桐 豊雅 : Involvement of proteasome-associated gene 1 (PAG1) in proliferation of triple negative breast cancer. (トリプルネガティブ乳癌におけるプロテアソーム関連遺伝子PAG1を介した発癌・増殖機構の関与), 第71回 日本癌学会学術総会, 2012年9月.
Le T. Dat, Taisuke Matsuo, Tetsuro Yoshimaru, Takuya Kuramoto, Masaki Hanibuchi, Hisatsugu Goto, Souji Kakiuchi, Yasuhiko Nishioka, Saburo Sone and Toyomasa Katagiri : Genome-wide gene expression profiling analysis of bone metastases of human non-smal cell lung cancer (NSCLC) in mice, 70th Annual Meeting of the Japanese Cancer Association, Oct. 2011.
172.
Komatsu Masato, Tetsuro Yoshimaru, Taisuke Matsuo, Miyoshi Yasuo, Mitsuo Shimada, Nakamura Yusuke, Sasa Mitsunori, Miyano Satoru and Toyomasa Katagiri : Novel therapeutic strategy for Triple Negative Breast Cancer, Proceedings of the Japanese Cancer Association, 237, Oct. 2011.
Dat LeTan, Taisuke Matsuo, Hisatsugu Goto, Souji Kakiuchi, Yasuhiko Nishioka, Saburo Sone and Toyomasa Katagiri : Identification and characterization of a novel transcription factor invoved lung cancer bone metastases, 第15回日本がん分子標的治療学会学術集会, Jun. 2011.
松尾 泰佑, 尾野 雅哉, 吉丸 哲郎, 三好 康雄, 笹 三徳, 中村 祐輔, 片桐 豊雅 : Identification of characterization of a novel glycosyltrasnferase BCGT1 involved in breast cancer cells, 平成23年度がん若手研究者ワークショップ, 2011年9月.