Shoko Matsushita, Kentaro Suzuki, Yukiko Ogino, Shinjiro Hino, Tetsuya Sato, Mikita Suyama, Takahiro Matsumoto, Akiko Omori, Satoshi Inoue and Gen Yamada : Androgen Regulates Mafb Expression Through its 3'UTR During Mouse Urethral Masculinization., Endocrinology, Vol.157, No.2, 844-857, 2015.
(要約)
External genitalia are prominent organs showing hormone-dependent sexual differentiation. Androgen is an essential regulator of masculinization of the genital tubercle, which is the anlage of external genitalia. We have previously shown that v-maf avian musculoaponeurotic fibrosarcoma oncogene homolog B (MAFB) is an androgen-inducible regulator of embryonic urethral masculinization in mice. However, it remains unclear how androgen regulates Mafb expression. The current study suggests that the Mafb 3' untranslated region (UTR) is an essential region for its regulation by androgen. We identified 2 functional androgen response elements (AREs) in Mafb 3'UTR. Androgen receptor is bound to such AREs in 3'UTR during urethral masculinization. In addition to 3'UTR, Mafb 5'UTR also showed androgen responsiveness. Moreover, we also demonstrated that -catenin, one of genital tubercle masculinization factors, may be an additional regulator of Mafb expression during urethral masculinization. This study provides insights to elucidate mechanisms of gene regulation through AREs present in Mafb 3'UTR for a better understanding of the processes of urethral masculinization.
Tai-yong Yu, Takeshi Kondo, Takahiro Matsumoto, Yoshiaki Fujii-Kuriyama and Yuuki Imai : Aryl hydrocarbon receptor catabolic activity in bone metabolism is osteoclast dependent in vivo., Biochemical and Biophysical Research Communications, Vol.450, No.1, 416-422, 2014.
(要約)
Bone mass is regulated by various molecules including endogenous factors as well as exogenous factors, such as nutrients and pollutants. Aryl hydrocarbon receptor (AhR) is known as a dioxin receptor and is responsible for various pathological and physiological processes. However, the role of AhR in bone homeostasis remains elusive because the cell type specific direct function of AhR has never been explored in vivo. Here, we show the cell type specific function of AhR in vivo in bone homeostasis. Systemic AhR knockout (AhRKO) mice exhibit increased bone mass with decreased resorption and decreased formation. Meanwhile, osteoclast specific AhRKO (AhR(Oc/Oc)) mice have increased bone mass with reduced bone resorption, although the mice lacking AhR in osteoblasts have a normal bone phenotype. Even under pathological conditions, AhR(Oc/Oc) mice are resistant to sex hormone deficiency-induced bone loss resulting from increased bone resorption. Furthermore, 3-methylcholanthrene, an AhR agonist, induces low bone mass with increased bone resorption in control mice, but not in AhR(Oc/Oc) mice. Taken together, cell type specific in vivo evidence for AhR functions indicates that osteoclastic AhR plays a significant role in maintenance of bone homeostasis, suggesting that inhibition of AhR in osteoclasts can be beneficial in the treatment of osteoporosis.
(キーワード)
Animals / Bone Density / Bone Resorption / Female / Femur / Male / 骨ミネラル代謝 (bone and mineral metabolism) / ノックアウトマウス (knockout mice) / Mice, Knockout / Organ Size / Osteoclasts / Ovariectomy / Receptors, Aryl Hydrocarbon
Sumiko Yoshida, Ken-ichi Aihara, Yasumasa Ikeda, Yuka Sumitomo-Ueda, Ryoko Uemoto, Kazue Ishikawa, Takayuki Ise, Shusuke Yagi, Takashi Iwase, Yasuhiro Mouri, Matomo Sakari, Takahiro Matsumoto, Ken-ichi Takeyama, Masashi Akaike, Mitsuru Matsumoto, Masataka Sata, Kenneth Walsh, Shigeaki Kato and Toshio Matsumoto : Androgen receptor promotes sex-independent angiogenesis in response to ischemia and is required for activation of vascular endothelial growth factor receptor signaling., Circulation, Vol.128, No.1, 60-71, 2013.
(要約)
These results document a physiological role of AR in sex-independent angiogenic potency and provide evidence of novel cross-talk between the androgen/AR signaling and VEGF/kinase insert domain protein receptor signaling pathways.
Yosuke Okuno, Fumiaki Ohtake, Katsuhide Igarashi, Jun Kanno, Takahiro Matsumoto, Ichiro Takada, Shigeaki Kato and Yuuki Imai : Epigenetic regulation of adipogenesis by PHF2 histone demethylase., Diabetes, Vol.62, No.5, 1426-1434, 2013.
(要約)
PHF2 is a JmjC family histone demethylase that removes the methyl group from H3K9me2 and works as a coactivator for several metabolism-related transcription factors. In this study, we examined the in vivo role of PHF2 in mice. We generated Phf2 floxed mice, systemic Phf2 null mice by crossing Phf2 floxed mice with CMV-Cre transgenic mice, and tamoxifen-inducible Phf2 knockout mice by crossing Phf2 floxed mice with Cre-ERT2 transgenic mice. Systemic Phf2 null mice had partial neonatal death and growth retardation and exhibited less adipose tissue and reduced adipocyte numbers compared with control littermates. Tamoxifen-induced conditional knockout of PHF2 resulted in impaired adipogenesis in stromal vascular cells from the adipose tissue of tamoxifen-inducible Phf2 knockout mice as well as of Phf2 knocked-down 3T3-L1 cells. PHF2 interacts with CEBPA and demethylates H3K9me2 in the promoters of CEBPA-regulated adipogenic genes. These findings suggest that PHF2 histone demethylase potentiates adipogenesis through interaction with CEBPA in vivo. Taken together, PHF2 may be a novel therapeutic target in the treatment of obesity and the metabolic syndrome.
Takahiro Matsumoto, Matomo Sakari, Maiko Okada, Atsushi Yokoyama, Sayuri Takahashi, Alexander Kouzmenko and Shigeaki Kato : The androgen receptor in health and disease., Annual Review of Physiology, Vol.75, 201-224, 2012.
(要約)
Androgens play pivotal roles in the regulation of male development and physiological processes, particularly in the male reproductive system. Most biological effects of androgens are mediated by the action of nuclear androgen receptor (AR). AR acts as a master regulator of downstream androgen-dependent signaling pathway networks. This ligand-dependent transcriptional factor modulates gene expression through the recruitment of various coregulator complexes, the induction of chromatin reorganization, and epigenetic histone modifications at target genomic loci. Dysregulation of androgen/AR signaling perturbs normal reproductive development and accounts for a wide range of pathological conditions such as androgen-insensitive syndrome, prostate cancer, and spinal bulbar muscular atrophy. In this review we summarize recent advances in understanding of the epigenetic mechanisms of AR action as well as newly recognized aspects of AR-mediated androgen signaling in both men and women. In addition, we offer a perspective on the use of animal genetic model systems aimed at eventually developing novel therapeutic AR ligands.
Sayuri Takahashi, Tomoyuki Watanabe, Maiko Okada, Kazuki Inoue, Takashi Ueda, Ichiro Takada, Tetsuro Watabe, Yoko Yamamoto, Toru Fukuda, Takashi Nakamura, Chihiro Akimoto, Tetsuya Fujimura, Maiko Hoshino, Yuuki Imai, Daniel Metzger, Kohei Miyazono, Yasuhiro Minami, Pierre Chambon, Tadaichi Kitamura, Takahiro Matsumoto and Shigeaki Kato : Noncanonical Wnt signaling mediates androgen-dependent tumor growth in a mouse model of prostate cancer., Proceedings of the National Academy of Sciences of the United States of America, Vol.108, No.12, 4938-4943, 2011.
(要約)
Prostate cancer development is associated with hyperactive androgen signaling. However, the molecular link between androgen receptor (AR) function and humoral factors remains elusive. A prostate cancer mouse model was generated by selectively mutating the AR threonine 877 into alanine in prostatic epithelial cells through Cre-ERT2-mediated targeted somatic mutagenesis. Such AR point mutant mice (ARpe-T877A/Y) developed hypertrophic prostates with responses to both an androgen antagonist and estrogen, although no prostatic tumor was seen. In prostate cancer model transgenic mice, the onset of prostatic tumorigenesis as well as tumor growth was significantly potentiated by introduction of the AR T877A mutation into the prostate. Genetic screening of mice identified Wnt-5a as an activator. Enhanced Wnt-5a expression was detected in the malignant prostate tumors of patients, whereas in benign prostatic hyperplasia such aberrant up-regulation was not obvious. These findings suggest that a noncanonical Wnt signal stimulates development of prostatic tumors with AR hyperfunction.
Aki Murashima, Shinichi Miyagawa, Yukiko Ogino, Hisayo Nishida-Fukuda, Kimi Araki, Takahiro Matsumoto, Takehito Kaneko, Kazuya Yoshinaga, Ken-ichi Yamamura, Takeshi Kurita, Shigeaki Kato, M Anne Moon and Gen Yamada : Essential roles of androgen signaling in Wolffian duct stabilization and epididymal cell differentiation., Endocrinology, Vol.152, No.4, 1640-1651, 2011.
(要約)
The epididymis is a male accessory organ and functions for sperm maturation and storage under the control of androgen. The development of the epididymis is also androgen dependent. The Wolffian duct (WD), anlagen of the epididymis, is formed in both male and female embryos; however, it is stabilized only in male embryos by testicular androgen. Androgen drives subsequent differentiation of the WD into the epididymis. Although the essential roles of androgen in WD masculinization and epididymal function have been established, little is known about cellular events regulated precisely by androgen signaling during these processes. It is also unclear whether androgen signaling, especially in the epithelia, has further function for epididymal epithelial cell differentiation. In this study we examined the cellular death and proliferation controlled by androgen signaling via the androgen receptor (AR) in WD stabilization. Analyses using AR knockout mice revealed that androgen signaling inhibits epithelial cell death in this process. Analysis of AP2α-Cre;AR(flox/Y) mice, in which AR function is deleted in the WD epithelium, revealed that epithelial AR is not required for the WD stabilization but is required for epithelial cell differentiation in the epididymis. Specifically, loss of epithelial AR significantly reduced expression of p63 that is essential for differentiation of basal cells in the epididymal epithelium. We also interrogated the possibility of regulation of the p63 gene (Trp63) by AR in vitro and found that p63 is a likely direct target of AR regulation.
Shinichi Miyagawa, Yoshihiko Satoh, Ryuma Haraguchi, Kentaro Suzuki, Taisen Iguchi, M Makoto Taketo, Naomi Nakagata, Takahiro Matsumoto, Ken-ichi Takeyama, Shigeaki Kato and Gen Yamada : Genetic interactions of the androgen and Wnt/beta-catenin pathways for the masculinization of external genitalia., Molecular Endocrinology, Vol.23, No.6, 871-880, 2009.
(要約)
In most mammals, the sexually dimorphic development of embryos is typically achieved by the differentiation of the external genitalia. Hence, the sexual distinction of mammalian newborns is based on the external genital structure. Although it was shown in the 1940s and 1950s that androgen from the testes establishes the male sexual characteristics, the involvement of nongonadal and locally produced masculine effectors remains totally unknown. It is noteworthy that the disorders of fetal masculinization, including hypospadias, one of the most frequent birth defects, occur at a high frequency. Furthermore, their causative factors remain unclear. In this study, the involvement of the coordinated actions of androgen and the growth factor systems was genetically analyzed for the first time on mammalian reproductive organ formation. The results demonstrated that the Wnt/beta-catenin pathway is indispensable masculine factor for the external genital development. The bilateral mesenchymal region adjacent to the urethral plate epithelium displayed a sexually dimorphic activity of Wnt/beta-catenin signaling. Loss- and gain-of-function beta-catenin mutants displayed altered sexual development of the external genitalia. These results indicate the novel functions of the Wnt/beta-catenin pathway as a locally expressed masculine effector. This could be the first genetic study analyzing the roles of the genetic interactions between androgen and locally expressed growth factor signaling during the development of reproductive organs. These results also shed new insight on the reproductive genetics and the causative factors of genital disorders.
Chihiro Akimoto, Hirochika Kitagawa, Takahiro Matsumoto and Shigeaki Kato : Spermatogenesis-specific association of SMCY and MSH5., Genes to Cells, Vol.13, No.6, 623-633, 2008.
(要約)
The status of chromatin during spermatogenesis is dynamically regulated by specific histone codes or stage-specific histone changes. The functional links between such epigenetic regulation and proteins regulating meiosis are largely unknown. In mammals, genes encoded on the Y chromosome are thought to possess male-specific biological functions. While genes located within the azoospermia factor region (AZF) are known to be involved in spermatogenesis, the physiological function of individual genes is not known. SMCY is a gene mapped to the AZF, and in this report, we analyzed the function of SMCY protein during spermatogenesis. Biochemical identification of the proteins with which it interacted showed that SMCY formed a distinct complex with MSH5, a critical meiosis-regulatory protein in the human testicular germ cell line, NEC8. As anticipated, histone H3K4 demethylase activity was detected. Immunohistochemical analysis revealed the co-localization of SMCY with MSH5 at a specific stage of meiotic prophase progression during murine spermatogenesis. Our results suggest that SMCY may have a male-specific function as a histone H3K4 demethylase by recruiting a meiosis-regulatory protein to condensed DNA.
Shingo Sato, Reiko Hanada, Ayako Kimura, Tomomi Abe, Takahiro Matsumoto, Makiko Iwasaki, Hiroyuki Inose, Takanori Ida, Michihiro Mieda, Yasuhiro Takeuchi, Seiji Fukumoto, Toshiro Fujita, Shigeaki Kato, Kenji Kangawa, Masayasu Kojima, Ken-ichi Shinomiya and Shu Takeda : Central control of bone remodeling by neuromedin U., Nature Medicine, Vol.13, No.10, 1234-1240, 2007.
(要約)
Bone remodeling, the function affected in osteoporosis, the most common of bone diseases, comprises two phases: bone formation by matrix-producing osteoblasts and bone resorption by osteoclasts. The demonstration that the anorexigenic hormone leptin inhibits bone formation through a hypothalamic relay suggests that other molecules that affect energy metabolism in the hypothalamus could also modulate bone mass. Neuromedin U (NMU) is an anorexigenic neuropeptide that acts independently of leptin through poorly defined mechanisms. Here we show that Nmu-deficient (Nmu-/-) mice have high bone mass owing to an increase in bone formation; this is more prominent in male mice than female mice. Physiological and cell-based assays indicate that NMU acts in the central nervous system, rather than directly on bone cells, to regulate bone remodeling. Notably, leptin- or sympathetic nervous system-mediated inhibition of bone formation was abolished in Nmu-/- mice, which show an altered bone expression of molecular clock genes (mediators of the inhibition of bone formation by leptin). Moreover, treatment of wild-type mice with a natural agonist for the NMU receptor decreased bone mass. Collectively, these results suggest that NMU may be the first central mediator of leptin-dependent regulation of bone mass identified to date. Given the existence of inhibitors and activators of NMU action, our results may influence the treatment of diseases involving low bone mass, such as osteoporosis.
(キーワード)
Absorptiometry, Photon / Animals / Bone Density / Bone Remodeling / 細胞増殖·分化 (cell proliferation and differentiation) / Cells, Cultured / Dose-Response Relationship, Drug / Female / 免疫組織化学 (immunohistochemistry) / Leptin / Male / Mice / Mice, Inbred C57BL / ノックアウトマウス (knockout mice) / Mice, Mutant Strains / Models, Biological / Neuropeptides / Osteoblasts / Sex Factors / Sympathetic Nervous System / Tomography, X-Ray Computed
Takashi Nakamura, Yuuki Imai, Takahiro Matsumoto, Shingo Sato, Kazusane Takeuchi, Katsuhide Igarashi, Yoshifumi Harada, Yoshiaki Azuma, Andree Krust, Yoko Yamamoto, Hiroshi Nishina, Shu Takeda, Hiroshi Takayanagi, Daniel Metzger, Jun Kanno, Kunio Takaoka, John T Martin, Pierre Chambon and Shigeaki Kato : Estrogen prevents bone loss via estrogen receptor alpha and induction of Fas ligand in osteoclasts., Cell, Vol.130, No.5, 811-823, 2007.
(要約)
Estrogen prevents osteoporotic bone loss by attenuating bone resorption; however, the molecular basis for this is unknown. Here, we report a critical role for the osteoclastic estrogen receptor alpha (ERalpha) in mediating estrogen-dependent bone maintenance in female mice. We selectively ablated ERalpha in differentiated osteoclasts (ERalpha(DeltaOc/DeltaOc)) and found that ERalpha(DeltaOc/DeltaOc) females, but not males, exhibited trabecular bone loss, similar to the osteoporotic bone phenotype in postmenopausal women. Further, we show that estrogen induced apoptosis and upregulation of Fas ligand (FasL) expression in osteoclasts of the trabecular bones of WT but not ERalpha(DeltaOc/DeltaOc) mice. The expression of ERalpha was also required for the induction of apoptosis by tamoxifen and estrogen in cultured osteoclasts. Our results support a model in which estrogen regulates the life span of mature osteoclasts via the induction of the Fas/FasL system, thereby providing an explanation for the osteoprotective function of estrogen as well as SERMs.
Heparin cofactor II (HCII) specifically inhibits thrombin action at sites of injured arterial wall, and patients with HCII deficiency exhibit advanced atherosclerosis. However, the in vivo effects and the molecular mechanism underlying the action of HCII during vascular remodeling remain elusive. To clarify the role of HCII in vascular remodeling, we generated HCII-deficient mice by gene targeting. In contrast to a previous report, HCII(-/-) mice were embryonically lethal. In HCII(+/-) mice, prominent intimal hyperplasia with increased cellular proliferation was observed after tube cuff and wire vascular injury. The number of protease-activated receptor-1-positive (PAR-1-positive) cells was increased in the thickened vascular wall of HCII(+/-) mice, suggesting enhanced thrombin action in this region. Cuff injury also increased the expression levels of inflammatory cytokines and chemokines in the vascular wall of HCII(+/-) mice. The intimal hyperplasia in HCII(+/-) mice with vascular injury was abrogated by human HCII supplementation. Furthermore, HCII deficiency caused acceleration of aortic plaque formation with increased PAR-1 expression and oxidative stress in apoE-KO mice. These results demonstrate that HCII protects against thrombin-induced remodeling of an injured vascular wall by inhibiting thrombin action and suggest that HCII is potentially therapeutic against atherosclerosis without causing coagulatory disturbance.
(キーワード)
Animals / Base Sequence / Blood Vessels / DNA Primers / Embryo Loss / Female / Gene Targeting / Genes, Lethal / Genotype / Heparin Cofactor II / Heterozygote / Homozygote / Male / Mice / Mice, Inbred C57BL / ノックアウトマウス (knockout mice) / Pregnancy
Junko Miyamoto, Takahiro Matsumoto, Hiroko Shiina, Kazuki Inoue, Ichiro Takada, Saya Ito, Johbu Itoh, Takeo Minematsu, Takashi Sato, Toshihiko Yanase, Hajime Nawata, R Yoshiyuki Osamura and Shigeaki Kato : The pituitary function of androgen receptor constitutes a glucocorticoid production circuit., Molecular and Cellular Biology, Vol.27, No.13, 4807-4814, 2007.
(要約)
Androgen receptor (AR) mediates diverse androgen actions, particularly reproductive processes in males and females. AR-mediated androgen signaling is considered to also control metabolic processes; however, the molecular basis remains elusive. In the present study, we explored the molecular mechanism of late-onset obesity in male AR null mutant (ARKO) mice. We determined that the obesity was caused by a hypercorticoid state. The negative feedback system regulating glucocorticoid production was impaired in ARKO mice. Male and female ARKO mice exhibited hypertrophic adrenal glands and glucocorticoid overproduction, presumably due to high levels of adrenal corticotropic hormone. The pituitary glands of the ARKO males had increased expression of proopiomelanocortin and decreased expression of the glucocorticoid receptor (GR). There were no overt structural abnormalities and no alteration in the distribution of cell types in the pituitaries of male ARKO mice. Additionally, there was normal production of the other hormones within the glucocorticoid feedback system in both the pituitary and hypothalamus. In a cell line derived from pituitary glands, GR expression was under the positive control of the activated AR. Thus, this study suggests that the activated AR supports the negative feedback regulation of glucocorticoid production via up-regulation of GR expression in the pituitary gland.
Shuhei Kimura, Takahiro Matsumoto, Reiko Matsuyama, Hiroko Shiina, Takashi Sato, Ken-Ichi Takeyama and Shigeaki Kato : Androgen receptor function in folliculogenesis and its clinical implication in premature ovarian failure., Trends in Endocrinology and Metabolism, Vol.18, No.5, 183-189, 2007.
(要約)
The action of estrogen in the female reproductive organs is well known in terms of the expression pattern and gene regulation of the estrogen receptor (ER). The significance of ERs in female reproduction is undisputed. The role of the androgen receptor (AR) is less clear. Clinical hyperandrogenism, a typical feature of polycystic ovary syndrome (PCOS), highlights pathological androgen production by the ovary. By contrast, the physiological impact of androgen action in female reproductive organs remains elusive. Androgens affect folliculogenesis in a variety of experimental approaches and ARs are expressed in developing follicles. Recent observations have discovered that inactivation of ARs in female mice results in premature ovarian failure (POF), indicating that normal folliculogenesis requires AR-mediated androgen action. Moreover, these results imply that POF might be caused by impairment of AR-mediated androgen action.
Hiroko Shiina, Takahiro Matsumoto, Takashi Sato, Katsuhide Igarashi, Junko Miyamoto, Sayuri Takemasa, Matomo Sakari, Ichiro Takada, Takashi Nakamura, Daniel Metzger, Pierre Chambon, Jun Kanno, Hiroyuki Yoshikawa and Shigeaki Kato : Premature ovarian failure in androgen receptor-deficient mice., Proceedings of the National Academy of Sciences of the United States of America, Vol.103, No.1, 224-229, 2005.
(要約)
Premature ovarian failure (POF) syndrome, an early decline of ovarian function in women, is frequently associated with X chromosome abnormalities ranging from various Xq deletions to complete loss of one of the X chromosomes. However, the genetic locus responsible for the POF remains unknown, and no candidate gene has been identified. Using the Cre/LoxP system, we have disrupted the mouse X chromosome androgen receptor (Ar) gene. Female AR(-/-) mice appeared normal but developed the POF phenotype with aberrant ovarian gene expression. Eight-week-old female AR(-/-) mice are fertile, but they have lower follicle numbers and impaired mammary development, and they produce only half of the normal number of pups per litter. Forty-week-old AR(-/-) mice are infertile because of complete loss of follicles. Genome-wide microarray analysis of mRNA from AR(-/-) ovaries revealed that a number of major regulators of folliculogenesis were under transcriptional control by AR. Our findings suggest that AR function is required for normal female reproduction, particularly folliculogenesis, and that AR is a potential therapeutic target in POF syndrome.
Kimiko Yamamoto, Takaaki Sokabe, Takahiro Matsumoto, Kimihiro Yoshimura, Masahiro Shibata, Norihiko Ohura, Toru Fukuda, Takashi Sato, Keisuke Sekine, Shigeaki Kato, Masashi Isshiki, Toshiro Fujita, Mikio Kobayashi, Koichi Kawamura, Hirotake Masuda, Akira Kamiya and Joji Ando : Impaired flow-dependent control of vascular tone and remodeling in P2X4-deficient mice., Nature Medicine, Vol.12, No.1, 133-137, 2005.
(要約)
The structure and function of blood vessels adapt to environmental changes such as physical development and exercise. This phenomenon is based on the ability of the endothelial cells to sense and respond to blood flow; however, the underlying mechanisms remain unclear. Here we show that the ATP-gated P2X4 ion channel, expressed on endothelial cells and encoded by P2rx4 in mice, has a key role in the response of endothelial cells to changes in blood flow. P2rx4(-/-) mice do not have normal endothelial cell responses to flow, such as influx of Ca(2+) and subsequent production of the potent vasodilator nitric oxide (NO). Additionally, vessel dilation induced by acute increases in blood flow is markedly suppressed in P2rx4(-/-) mice. Furthermore, P2rx4(-/-) mice have higher blood pressure and excrete smaller amounts of NO products in their urine than do wild-type mice. Moreover, no adaptive vascular remodeling, that is, a decrease in vessel size in response to a chronic decrease in blood flow, was observed in P2rx4(-/-) mice. Thus, endothelial P2X4 channels are crucial to flow-sensitive mechanisms that regulate blood pressure and vascular remodeling.
Shigeaki Kato, Takashi Sato, Tomoyuki Watanabe, Sayuri Takemasa, Yoshikazu Masuhiro, Fumiaki Ohtake and Takahiro Matsumoto : Function of nuclear sex hormone receptors in gene regulation., Cancer Chemotherapy and Pharmacology, Vol.56 Suppl 1, 4-9, 2005.
(要約)
The development of reproductive organ tumors such as breast and prostate cancer often depends on the action of sex hormones. Nuclear sex hormone receptors are members of the nuclear hormone receptor superfamily and act as ligand-inducible transcription factors, controlling the expression of target genes. Nuclear receptors are considered to directly and indirectly interact with a number of nuclear co-regulatory complexes involved in chromatin remodeling and histone modification. Moreover, many intracellular signalings via cell membrane receptors are shown to modulate nuclear receptor-regulated transcription. We have shown that estrogen receptors (ER) associate with a number of nuclear complexes, one of which is a spliceosome complex. We recently found that this spliceosome complex interacts with phosphorylated ER by MAP kinase, generating a novel cross-talk of estrogen and growth factor signalings. We also observed that a dioxin receptor (AhR) is capable of associating with ER, resulting in modulation of ER transactivation function. From our findings we believe that development of estrogen-dependent breast cancer may be mediated through the other signaling pathways. To address the function of the androgen receptor (AR) in androgen-dependent prostate cancer, we established a transgenic mouse line expressing a human AR mutant that is found in androgen-independent prostate cancer patients. The hAR mutant mice, generated through a Cre-loxP system, developed hyperplasia in the prostates. Hypersensitivity of AR mutants to antagonists and endogenous steroid hormones may potentiate hormone-dependency in prostate cancer development.
Takahiro Matsumoto, Ken-ichi Takeyama, Takashi Sato and Shigeaki Kato : Study of androgen receptor functions by genetic models., The Journal of Biochemistry, Vol.138, No.2, 105-110, 2005.
(要約)
Androgens exert most of their biological activities through binding to the androgen receptor (AR). The AR belongs to the nuclear receptor superfamily and acts as a ligand-inducible transcriptional factor. AR dysfunction causes a diverse range of clinical conditions, such as testicular mutation (Tfm) syndrome, prostate cancer, and spinal and bulbar muscular atrophy (SBMA). However, the molecular basis of the AR function underlying these AR-related disorders remains largely unknown due to the lack of stable genetic models. Here we review recent results of our studies into genetic models of the loss of AR function in mice and the gain of AR function in Drosophila.
Androgen has anabolic effects on cardiac myocytes and has been shown to enhance left ventricular enlargement and function. However, the physiological and patho-physiological roles of androgen in cardiac growth and cardiac stress-induced remodeling remains unclear. We aimed to clarify whether the androgen-nuclear androgen receptor (AR) system contributes to the cardiac growth and angiotensin II (Ang II)-stimulated cardiac remodeling by using systemic AR-null male mice. AR knock-out (ARKO) male mice, at 25 weeks of age, and age-matched wild-type (WT) male mice were treated with or without Ang II stimulation (2.0 mg/kg/day) for 2 weeks. ARKO mice with or without Ang II stimulation showed a significant reduction in the heart-to-body weight ratio compared with those of WT mice. In addition, echocardiographic analysis demonstrated impairments of both the concentric hypertrophic response and left ventricular function in Ang II-stimulated ARKO mice. Western blot analysis of the myocardium revealed that activation of extracellular signal-regulated kinases (ERK) 1/2 and ERK5 by Ang II stimulation were lower in ARKO mice than those of WT mice. Ang II stimulation caused more prominent cardiac fibrosis in ARKO mice than in WT mice with enhanced expression of types I and III collagen and transforming growth factor-beta1 genes and with increased Smad2 activation. These results suggest that, in male mice, the androgen-AR system participates in normal cardiac growth and modulates cardiac adaptive hypertrophy and fibrosis during the process of cardiac remodeling under hypertrophic stress.
Takashi Yamada, Hirotaka Kawano, Keisuke Sekine, Takahiro Matsumoto, Toru Fukuda, Yoshiaki Azuma, Keiji Itaka, Ung-il Chung, Pierre Chambon, Kozo Nakamura, Shigeaki Kato and Hiroshi Kawaguchi : SRC-1 is necessary for skeletal responses to sex hormones in both males and females., Journal of Bone and Mineral Research, Vol.19, No.9, 1452-1461, 2004.
(要約)
Although SRC-1(-/-) mice showed no abnormality in growth or major organs, both males and females showed osteopenia with high bone turnover in the trabecular bones, but not in the cortical bones, compared with WT littermates. Their serum levels of sex hormones were upregulated, suggesting a compensatory reaction for the insensitivity to these hormones. Gonadectomies caused decreases in BMDs of SRC-1(-/-) and WT mice to the same levels; however, replacement with 5 alpha-dihydrotestosterone and 17 beta-estradiol in males and females, respectively, failed to restore the bone loss in SRC-1(-/-), whereas the WT bone volume was increased to the sham-operated levels. In contrast, bone loss by administered prednisolone was similarly seen in SRC-1(-/-) and WT mice. We conclude that SRC-1 is essential for the maintenance of bone mass by sex hormones, but not for the catabolic action of glucocorticoid, under both physiological and pathological conditions.
Shigeaki Kato, Takahiro Matsumoto, Hirotaka Kawano, Takashi Sato and Ken-ichi Takeyama : Function of androgen receptor in gene regulations., The Journal of Steroid Biochemistry and Molecular Biology, Vol.89-90, No.1-5, 627-633, 2004.
(要約)
Most of the androgen actions are considered to be mediated by the androgen receptor (AR) of the target genes. The AR is composed of a fairly large molecule because of the long A/B domains of its N-terminal. However, the independent roles of the AR as well as those of the estrogen receptors largely remained unknown mainly due to the lack of the AR knockout (ARKO) mice line. We have succeeded in generating the ARKO mouse by means of a conditional targeting using the Cre/loxP system. The ARKO males grew healthily although they showed a typical feature of the testicular feminization mutation (Tfm) and the hormonal assay revealed significantly lower serum androgen and higher LH levels in comparison with those of the wild type (WT) males. The serum estrogen levels were, however, comparable between both the ARKO and the WT. Another hallmark of the ARKO males was a state of high bone turnover osteopenia, in which the acceleration in the bone resorption clearly exceeded the bone formation. Male-typical behaviors were disrupted in male ARKO mice. Aiming at a quick differentiation of an androgen-dependent polyQ disease such as Kennedy's disease, the authors also developed the Drosophila fly-eye model in which the wild type and the polyQ-expanded human AR (hAR) was induced in the eyes of Drosophila. When androgen was administered to the flies induced with the polyQ-expanded hAR, their optical nerves were devastated.
Takashi Sato, Takahiro Matsumoto, Hirotaka Kawano, Tomoyuki Watanabe, Yoshikatsu Uematsu, Keisuke Sekine, Toru Fukuda, Ken-ichi Aihara, Andrée Krust, Takashi Yamada, Yuko Nakamichi, Yoko Yamamoto, Takashi Nakamura, Kimihiro Yoshimura, Tatsuya Yoshizawa, Daniel Metzger, Pierre Chambon and Shigeaki Kato : Brain masculinization requires androgen receptor function., Proceedings of the National Academy of Sciences of the United States of America, Vol.101, No.6, 1673-1678, 2004.
(要約)
Testicular testosterone produced during a critical perinatal period is thought to masculinize and defeminize the male brain from the inherent feminization program and induce male-typical behaviors in the adult. These actions of testosterone appear to be exerted not through its androgenic activity, but rather through its conversion by brain aromatase into estrogen, with the consequent activation of estrogen receptor (ER)-mediated signaling. Thus, the role of androgen receptor (AR) in perinatal brain masculinization underlying the expression of male-typical behaviors remains unclear because of the conversion of testosterone into estrogen in the brain. Here, we report a null AR mutation in mice generated by the Cre-loxP system. The AR-null mutation in males (AR(L-/Y)) resulted in the ablation of male-typical sexual and aggressive behaviors, whereas female AR-null homozygote (AR(L-/L-)) mice exhibited normal female sexual behaviors. Treatment with nonaromatizable androgen (5alpha-dihydrotestosterone, DHT) was ineffective in restoring the impaired male sexual behaviors, but it partially rescued impaired male aggressive behaviors in AR(L-/Y) mice. Impaired male-typical behaviors in ERalpha(-/-) mice were restored on DHT treatment. The role of AR function in brain masculinization at a limited perinatal stage was studied in AR(L-/L-) mice. Perinatal DHT treatment of females led to adult females sensitive to both 17beta-estradiol and DHT in the induction of male-typical behaviors. However, this female brain masculinization was abolished by AR inactivation. Our results suggested that perinatal brain masculinization requires AR function and that expression of male-typical behaviors in adults is mediated by both AR-dependent and -independent androgen signaling.
Hirotaka Kawano, Takashi Sato, Takashi Yamada, Takahiro Matsumoto, Keisuke Sekine, Tomoyuki Watanabe, Takashi Nakamura, Toru Fukuda, Kimihiro Yoshimura, Tatsuya Yoshizawa, Ken-ichi Aihara, Yoko Yamamoto, Yuko Nakamichi, Daniel Metzger, Pierre Chambon, Kozo Nakamura, Hiroshi Kawaguchi and Shigeaki Kato : Suppressive function of androgen receptor in bone resorption., Proceedings of the National Academy of Sciences of the United States of America, Vol.100, No.16, 9416-9421, 2003.
(要約)
As locally converted estrogen from testicular testosterone contributes to apparent androgen activity, the physiological significance of androgen receptor (AR) function in the beneficial effects of androgens on skeletal tissues has remained unclear. We show here that inactivation of AR in mice using a Cre-loxP system-mediated gene-targeting technique caused bone loss in males but not in females. Histomorphometric analyses of 8-week-old male AR knockout (ARKO) mice showed high bone turnover with increased bone resorption that resulted in reduced trabecular and cortical bone mass without affecting bone shape. Bone loss in orchidectomized male ARKO mice was only partially prevented by treatment with aromatizable testosterone. Analysis of primary osteoblasts and osteoclasts from ARKO mice revealed that AR function was required for the suppressive effects of androgens on osteoclastogenesis supporting activity of osteoblasts but not on osteoclasts. Furthermore, expression of the receptor activator of NF-kappaB ligand (RANKL) gene, which encodes a major osteoclastogenesis inducer, was found to be up-regulated in osteoblasts from AR-deficient mice. Our results indicate that AR function is indispensable for male-type bone formation and remodeling.
(キーワード)
Androgen-Insensitivity Syndrome / Animals / Bone Diseases, Metabolic / Bone Resorption / Carrier Proteins / Female / Male / Membrane Glycoproteins / Mice / ノックアウトマウス (knockout mice) / Mutation / Osteoblasts / Osteoclasts / RANK Ligand / Receptor Activator of Nuclear Factor-kappa B / Receptors, Androgen / Reverse Transcriptase Polymerase Chain Reaction / Sex Factors / Time Factors / Up-Regulation
Takahiro Matsumoto, Ken-ichi Takeyama, Takashi Sato and Shigeaki Kato : Androgen receptor functions from reverse genetic models., The Journal of Steroid Biochemistry and Molecular Biology, Vol.85, No.2-5, 95-99, 2003.
(要約)
The androgen receptor (AR) is a ligand-dependent transcription factor involved in the regulation of many different physiological processes. AR dysfunction causes a diverse range of clinical conditions, including testicular feminization mutation (Tfm) syndrome, prostate cancer, and motor neuron disease (Kennedy's disease). However, due to lack of genetic models, the molecular basis of the AR in these disorders remains largely unknown. Using a conditional targeting technique based on the Cre-loxP system, we successfully generated null AR mutant (ARKO) mice. ARKO males exhibited normal healthy growth, but showed typical Tfm abnormalities. Hormonal assay of ARKO males revealed that while serum androgen levels were very low, estrogen levels were normal. Another hallmark of ARKO males was late-onset obesity, with marked accumulation of white adipose tissue. To clarify the role of human AR (hAR) mutants with expanded polyQ stretches as observed in neurodegenerative disease, we also established a Drosophila model in which either wild-type or polyQ-expanded hAR were ectopically expressed. Although no overt phenotype was detected in adult fly-eye neurons expressing mutant hAR, the ingestion of androgen caused marked neurodegeneration.
Takashi Sato, Takahiro Matsumoto, Takashi Yamada, Tomoyuki Watanabe, Hirotaka Kawano and Shigeaki Kato : Late onset of obesity in male androgen receptor-deficient (AR KO) mice., Biochemical and Biophysical Research Communications, Vol.300, No.1, 167-171, 2003.
(要約)
An androgen receptor (AR) null mutant mice line was generated by means of a Cre-lox P system. The male (AR(L-/Y)) (KO) mice exhibited typical features of testicular feminization mutant (Tfm) disease in external reproductive organs with growth retardation. The growth curve of the male AR KO mice was similar to that of the wild-type female littermates until the 10th week of age, but thereafter a drastic increase in the growth was observed with development of obesity. Clear increase in the wet weights of white adipose tissues, but not of brown adipose tissue, was found in the 30-week-old male AR KO mice. However, no significant alteration in serum lipid parameters and food intake was observed. Thus, the present results suggest that AR may serve as a negative regulator of adipose development in adult males.
Takahiro Matsumoto, Kazuki Inoue, Takashi Sato and Shigeaki Kato : Genetic impact of both sex hormones in male-typical behaviors., Advances in Experimental Medicine and Biology, Vol.707, 125-126, 2011.
Yuuki Imai, Ming-Young Youn, Shino Kondoh, Takashi Nakamura, Alexander Kouzmenko, Takahiro Matsumoto, Ichiro Takada, Kunio Takaoka and Shigeaki Kato : Estrogens maintain bone mass by regulating expression of genes controlling function and life span in mature osteoclasts., Annals of the New York Academy of Sciences, Vol.1173 Suppl 1, E31-9, 2009.
(要約)
Estrogens play a key role in regulation of bone mass and strength by controlling activity of bone-forming osteoblasts and bone-resorbing osteoclasts. Cellular effects of estrogens are mediated predominantly by the action of estrogen receptor alpha (ERalpha). In earlier studies, ablation of the ERalpha gene in mice did not result in osteoporotic phenotypes due to systemic endocrine disturbance and compensatory effects of elevated levels of testosterone. Despite the relatively well-established effects in osteoblasts, little is known about the direct action of estrogen in osteoclasts. Development in the last decade of more sophisticated genetic manipulation approaches opened new possibilities to explore cell-specific roles of nuclear receptors in bone tissue. Recently, we have generated osteoclast-specific ERalpha gene knockout mice and shown that in vivo estrogens directly regulate the life span of mature osteoclasts by inducing the expression of pro-apoptotic Fas ligand (FasL). Inhibitory effects of estrogens on osteoclast function were further studied in vitro. We observed sufficiently detectable ERalpha expression in osteoclasts differentiating from primary bone marrow cells or RAW264 cells, although levels of ERalpha were decreasing during progression of the differentiation into mature osteoclasts. Treatment with estrogens led to reduction in expression of osteoclast-specific genes controlling bone resorption activity. However, estrogens did not affect the size of multinucleated osteoclasts or number of nuclei in a mature osteoclast. In conclusion, in osteoclasts, estrogens function to inhibit bone resorption activity and vitality rather than differentiation.
Takahiro Matsumoto, Hiroko Shiina, Hirotaka Kawano, Takashi Sato and Shigeaki Kato : Androgen receptor functions in male and female physiology., The Journal of Steroid Biochemistry and Molecular Biology, Vol.109, No.3-5, 236-241, 2008.
(要約)
The biological actions of androgens are mediated by transcriptional control of target genes through binding to the androgen receptor (AR). The AR belongs to the nuclear receptor superfamily and acts as a ligand-inducible transcriptional factor. Androgens/AR system is thought to be central to male physiology and behaviors to the development of clinical pathology. The physiological importance of AR function currently recognized in female reproduction also has been verified in mouse genetic model. Here we review functions of AR in male and female physiology as revealed by mice lacking AR.
(キーワード)
Animals / Bone Diseases, Metabolic / Female / Humans / Male / 肥満症 (obesity) / Phenotype / Protein Binding / Receptors, Androgen / Sex Characteristics
Yuuki Imai, Takashi Nakamura, Takahiro Matsumoto, Kunio Takaoka and Shigeaki Kato : Molecular mechanisms underlying the effects of sex steroids on bone and mineral metabolism., Journal of Bone and Mineral Metabolism, Vol.27, No.2, 127-130, Jan. 2009.
(要約)
The multifarious functions of sex steroid receptors include a role as regulatory factors for bone and mineral metabolism in vivo. These functions are more complex than originally assumed. The finding of nuclear receptors in osseous tissue alludes to the existence of novel indirect and direct functions of bone tissue beyond skeletal support, hematopoiesis, and calcium homeostasis. Cell-specific gene targeting approaches are an extremely important technology for future studies that will need to be conducted to fully understand the molecular mechanisms underlying bone formation and metabolism.
Takahiro Matsumoto, Masayoshi Yonezawa and Matomo Sakari : Male-specific histone acetylation through a cooperation between Y chromosomal protein CDY and androgen signaling, Nuclear Receptors: Biological Networks, Genome Dynamics and Disease, Keystone Symposia, Jan. 2014.
2.
Matomo Sakari and Takahiro Matsumoto : Y Chromosomal Azoospermic Factor RBMY and Estrogen Signaling Coordinately Regulate Male-specific Alternative Splicing, 15th International Congress on Hormonal Steroids and Hormones & Cancer, Kanazawa, Nov. 2012.
3.
Takahiro Matsumoto : Non-canonical Wnt signaling links AR mutation to prostate cancer, 15th International Congress on Hormonal Steroids and Hormones & Cancer, Kanazawa, Nov. 2012.
4.
Shigeaki Kato and Takahiro Matsumoto : A Molecular Link of Steroid Receptor Function to Cell Proliferation, 15th International Congress on Hormonal Steroids and Hormones & Cancer, Kanazawa, Nov. 2012.
5.
Matomo Sakari and Takahiro Matsumoto : Y chromosomal azoospermic factor RBMY and estrogen signaling coordinately regulate male-specific alternative splicing, The 58th/60th NIBB Conference Germline-Specification, Sex and Stem Cells-, Okazaki, Jul. 2012.
6.
Takahiro Matsumoto : Androgen receptor functions in male and female physiology & behavior, 6th JAPAN-ASEAN Conference on Men's Health & Aging, Jul. 2011.
松本 高広, 米澤 正祥, 井上 和樹, 盛 真友 : Male-specific histone acetylation trough a cooperation between Y chromosomal Protein CDY and androgen signaling, 第35回日本分子生物学会年会, 2012年12月.