Tatsuya Tominaga, Hideharu Abe, Akira Mima, Kojiro Nagai, Matsubara Takeshi and Toshio Doi : Role of Smad 1 Signaling in Pathogenesis of Diabetic Nephropathy, Nova Science Publishers, Sep. 2012.
2.
冨永 辰也, 土井 俊夫 : 糖尿病性腎症のバイオマーカー開発と臨床への展望, 2011年8月.
3.
冨永 辰也, 土井 俊夫 : 糖尿病関連遺伝子改変マウス, 2010年8月.
学術論文(審査論文):
1.
Kenji Nishimura, Taichi Murakami, Toshihiro Sakurai, Masashi Miyoshi, Kiyoe Kurahashi, Seiji Kishi, Masanori Tamaki, Tatsuya Tominaga, Sumiko Yoshida, Kojiro Nagai, Hideharu Abe, Shu-Ping Hui, Kazuhiko Kotani and Toshio Doi : Circulating Apolipoprotein L1 is associated with insulin resistance-induced abnormal lipid metabolism., Scientific Reports, Vol.9, No.1, 14869, 2019.
(要約)
Circulating ApolipoproteinL1 (ApoL1) is a component of pre-β-high-density lipoprotein (HDL), however little is known about the relationship of ApoL1 with cardiometabolic factors. Considering previous studies reporting the correlation of ApoL1 to triglyceride, we have hypothesized that ApoL1 associates with insulin-related metabolism. The current study examined their associations in 126 non-diabetic subjects and 36 patients with type 2 diabetes (T2DM). Non-diabetic subjects demonstrated triglyceride (standardized coefficients [s.c.] = 0.204, p < 0.05), body mass index (s.c. =0.232, p < 0.05) and HDL cholesterol (s.c. = -0.203, p < 0.05) as independent determinant of ApoL1 levels, and the significant elevation of ApoL1 in metabolic syndrome. Lipoprotein fractionation analysis revealed the predominant distribution of ApoL1 in large HDL fraction, and the significant increase of ApoL1 in large LDL fraction in high ApoL1 samples with insulin resistance. In T2DM, ApoL1 was higher in T2DM with metabolic syndrome, however ApoL1 was lower with β cell dysfunction. Insulin significantly promotes ApoL1 synthesis and secretion in HepG2 cells. In conclusion, circulating ApoL1 may be associated with abnormal HDL metabolism in insulin resistant status. This may suggest a regulation of insulin signal on the ApoL1 level, leading to offer a novel insight to the ApoL1 biology.
Akiko Sakurai, Hiroyuki Ono, Arisa Ochi, Motokazu Matsuura, Sakiya Yoshimoto, Seiji Kishi, Taichi Murakami, Tatsuya Tominaga, Kojiro Nagai, Hideharu Abe and Toshio Doi : Involvement of Elf3 on Smad3 activation-dependent injuries in podocytes and excretion of urinary exosome in diabetic nephropathy., PLoS ONE, Vol.14, No.5, 2019.
(要約)
Diabetic nephropathy (DN) is among the most serious complications of diabetes mellitus, and often leads to end-stage renal disease ultimately requiring dialysis or renal transplantation. The loss of podocytes has been reported to have a role in the onset and progression of DN. Here, we addressed the activation mechanism of Smad3 signaling in podocytes. Expression of RII and activation of Smad3 were induced by AGE exposure (P<0.05). Reduction of the activation of RII-Smad3 signaling ameliorated podocyte injuries in Smad3-knockout diabetic mice. The bone morphogenetic protein 4 (BMP4) significantly regulated activation of RII-Smad3 signalings (P<0.05). Moreover, the epithelium-specific transcription factor, Elf3was induced by AGE stimulation and, subsequently, upregulated RII expression in cultured podocytes. Induction of Elf3 and activation of RII-Smad3 signaling, leading to a decrease in WT1 expression, were observed in podocytes in diabetic human kidneys. Moreover, AGE treatment induced the secretion of Elf3-containing exosomes from cultured podocytes, which was dependent on the activation of the TGF-β-Smad3 signaling pathway. In addition, exosomal Elf3 protein in urine could be measured only in urinary exosomes from patients with DN. The appearance of urinary exosomal Elf3 protein in patients with DN suggested the existence of irreversible injuries in podocytes. The rate of decline in the estimated Glomerular Filtration Rate (eGFR) after measurement of urinary exosomal Elf3 protein levels in patients with DN (R2 = 0.7259) might be useful as an early non-invasive marker for podocyte injuries in DN.
藤田 結衣, 冨永 辰也, 寒川 裕未, 長井 幸二郎, 安部 秀斉, 土井 俊夫 : BMP4 regulates both podocyte injury and mesangial expansion in the diabetic nephropathy, 四国医学雑誌, Vol.75, No.1-2, 55-62, 2019年.
(要約)
Podocyte injury and loss have been indicated as constituting the crucial pathogenesis of glomerular injury ; however, it remains necessary to elucidate the detailed molecular mechanisms and cell-to-cell response because multiple factors may cause podocyte injury. In the glomerulus, three kinds of cells (endothelial, mesangial, and parietal epithelial) react to podocyte injury. Endothelial and mesangial cells are connected with podocyte cells across the glomerular basement membrane. However, the detailed mechanisms regarding the interaction of the mesangium and podocyte injury are unclear. Diabetic nephropathy is characterized by mesangial matrix expansion caused by an excessive deposition of extracellular matrix proteins in the mesangial area, which can be observed through the increased expression of type IV collagen. We have shown that bone morphogenetic protein 4 (BMP4) signaling leads to the glomerular changes characteristic of this disorder. To analyze the effect of BMP4 was investigated in vitro and in vivo using streptozotocin (STZ)-induced and Bmp4 heterozygous knockout (Bmp4+/-) diabetic mice or podocyte-specific Bmp4 knockout mice, and Bmp4-induced or podocyte-specific transgenic mice. BMP4 positive area and mesangial area fraction showed positively correlation. Furthermore, mesangial area fraction was significantly and negatively correlated with,WT1-positive cell number, and nephrin-positive area. We also demonstrated that the induction of podocyte apoptosis by BMP4 may be mediated by p38 activation and that of caspase 3 through cleavage. In mesangial cells, BMP4 stimulation also induced phosphorylation of p38 and Smad1 and increased cleaved caspase 3, with similar significant inhibition of Smad1 activation and decreased cleaved caspase 3 mediated by dorsomorphin. These data suggest that the BMP4 signaling pathway plays important roles for the development of both podocyte injury and mesangial expansion in diabetic nephropathy.
Masanori Tamaki, Tatsuya Tominaga, Yui Fujita, Yasuhiko Koezuka, Go Ichien, Taichi Murakami, Seiji Kishi, Keiichi Yamamoto, Hideharu Abe, Kojiro Nagai and Toshio Doi : All-trans retinoic acid suppresses bone morphogenetic protein 4 in mouse diabetic nephropathy through a unique retinoic acid response element., American Journal of Physiology, Endocrinology and Metabolism, Vol.316, No.3, E418-E431, 2019.
(要約)
Diabetic nephropathy (DN) causes mesangial matrix expansion, which results in glomerulosclerosis and renal failure. Collagen IV (COL4) is a major component of the mesangial matrix that is positively regulated by bone morphogenetic protein 4 (BMP4)/suppressor of mothers against decapentaplegic (Smad1) signaling. Because previous studies showed that retinoids treatment had a beneficial effect on kidney disease, we investigated the therapeutic potential of retinoids in DN, focusing especially on the regulatory mechanism of BMP4. Diabetes was induced with streptozotocin in 12-wk-old male Crl:CD1(ICR) mice, and, 1 mo later, we initiated intraperitoneal injection of all-trans retinoic acid (ATRA) three times weekly. Glomerular matrix expansion, which was associated with increased BMP4, phosphorylated Smad1, and COL4 expression, worsened in diabetic mice at 24 wk of age. ATRA administration alleviated DN and downregulated BMP4, phosopho-Smad1, and COL4. In cultured mouse mesangial cells, treatment with ATRA or a retinoic acid receptor-α (RARα) agonist significantly decreased BMP4 and COL4 expression. Genomic analysis suggested two putative retinoic acid response elements (RAREs) for the mouse Bmp4 gene. Chromatin immunoprecipitation analysis and reporter assays indicated a putative RARE of the Bmp4 gene, located 11,488-11,501 bp upstream of exon 1A and bound to RARα and retinoid X receptor (RXR), which suppressed BMP4 expression after ATRA addition. ATRA suppressed BMP4 via binding of a RARα/RXR heterodimer to a unique RARE, alleviating glomerular matrix expansion in diabetic mice. These findings provide a novel regulatory mechanism for treatment of DN.
Tatsuya Tominaga, Isha Sharma, Yui Fujita, Toshio Doi, K Aryana Wallner and S Yashpal Kanwar : Myo-inositol oxygenase accentuates renal tubular injury initiated by endoplasmic reticulum stress., American Journal of Physiology, Renal Physiology, Vol.316, No.2, F301-F315, 2018.
(要約)
Besides oxidant stress, endoplasmic reticulum (ER) stress has been implicated in the pathogenesis of various metabolic disorders affecting the kidney. These two forms of stresses are not mutually exclusive to each other and may operate by a feedback loop in worsening the cellular injury. To attest to this contention, studies were performed to assess whether in such a setting, there is worsening of tubulointerstitial injury. We employed tunicamycin as a model of ER stress and used tubular cells and mice overexpressing myo-inositol oxygenase (MIOX), an enzyme involved in glycolytic events with excessive generation of ROS. Concomitant treatment of tunicamycin and transfection of cells with MIOX-pcDNA led to a marked generation of ROS, which was reduced by MIOX-siRNA. Likewise, an accentuated expression of ER stress sensors, GRP78, XBP1, and CHOP, was observed, which was reduced with MIOX-siRNA. These sensors were markedly elevated in MIOX-TG mice compared with WT treated with tunicamycin. This was accompanied with marked deterioration of tubular morphology, along with impairment of renal functions. Interestingly, minimal damage and elevation of ER stressors was observed in MIOX-KO mice. Downstream events that were more adversely affected in MIOX-TG mice included accentuated expression of proapoptogenic proteins, proinflammatory cytokines, and extracellular matrix constituents, although expression of these molecules was unaffected in MIOX-KO mice. Also, their tunicamycin-induced accentuated expression in tubular cells was notably reduced with MIOX-siRNA. These studies suggest that the biology of MIOX-induced oxidant stress and tunicamycin-induced ER stress are interlinked, and both of the events may feed into each other to amplify the tubulointerstitial injury.
Yui Fujita, Tatsuya Tominaga, Hideharu Abe, Yumi Kangawa, Naoshi Fukushima, Otoya Ueda, Kou-Ichi Jishage, Seiji Kishi, Taichi Murakami, Yumiko Saga, S Yashpal Kanwar, Kojiro Nagai and Toshio Doi : An adjustment in BMP4 function represents a treatment for diabetic nephropathy and podocyte injury., Scientific Reports, Vol.8, No.1, 2018.
(要約)
Podocyte injury has been proposed to play an important role in diabetic nephropathy; however, its pathological mechanism remains unclear. We have shown that bone morphogenetic protein 4 (BMP4) signaling leads to the glomerular changes characteristic of this disorder. To analyze the molecular mechanism of podocyte injury, the effect of BMP4 was investigated using streptozotocin (STZ)-induced, Bmp4 heterozygous knockout (Bmp4+/-) and podocyte-specific Bmp4 knockout mice. Mice with STZ-induced diabetes exhibited glomerular matrix hyperplasia and decreased numbers of podocyte nucleus-specific WT1-positive cells. The number of podocytes and proteinuria were improved in both diabetic Bmp4 knockout mouse models compared to the effects observed in the control mice. The effect of BMP4 overexpression on Bmp4-induced or podocyte-specific transgenic mice was examined. Tamoxifen-induced Bmp4-overexpressing mice exhibited mesangial matrix expansion and decreased numbers of WT1-positive cells. Podocyte-specific Bmp4-overexpressing mice displayed increased kidney BMP4 expression and mesangial matrix expansion but decreased nephrin expression and numbers of WT1-positive cells. Both lines of Bmp4-overexpressing mice exhibited increased albuminuria. In cultured podocytes, BMP4 increased phospho-p38 levels. BMP4 decreased nephrin expression but increased cleaved caspase-3 levels. p38 suppression inhibited caspase-3 activation. Apoptosis was confirmed in STZ-diabetic glomeruli and Bmp4-overexpressing mice. Bmp4 +/- mice with diabetes displayed reduced apoptosis. Based on these data, the BMP4 signaling pathway plays important roles in the development of both podocyte injury and mesangial matrix expansion in diabetic nephropathy.
Hiroyuki Ono, Hideharu Abe, Akiko Sakurai, Arisa Ochi, Tatsuya Tominaga, Masanori Tamaki, Seiji Kishi, Taichi Murakami, Kojiro Nagai, Masayuki Kohashi and Toshio Doi : Novel Interplay Between Smad1 and Smad3 Phosphorylation via AGE Regulates the Progression of Diabetic Nephropathy., Scientific Reports, Vol.8, No.1, 2018.
(要約)
; db/db mice and probucol-treated db/db mice, which was consistent with the attenuation of ECM overproduction. These results indicate that Smad3 expression and activation or probucol treatment alters Smad1 phosphorylation, thus suggesting new molecular mechanisms underlying DN development and progression.
Fumi Kishi, Kojiro Nagai, Norimichi Takamatsu, Tatsuya Tominaga, Masanori Tamaki, E. Shibata, Taichi Murakami, Seiji Kishi, Hideharu Abe, Y Koezuka, N Minagawa, G Ichien and Toshio Doi : Urinary type IV collagen excretion is involved in the decline in estimated glomerular filtration rate in the Japanese general population without diabetes: A 5-year observational study., PLoS ONE, Vol.13, No.4, e0195523, 2018.
(要約)
Urinary type IV collagen (U-Col4) and albumin excretion is evaluated to monitor the development of diabetic kidney disease. However, U-Col4 excretion in the general population without diabetes has not yet been fully elucidated. In this study, 1067 participants without diabetes and with urinary albumin-creatinine ratio <300 mg/gCr (normo- or microalbuminuria) who underwent an annual health examination in 2004 were enrolled and observed for 5 years. They were divided according to the amount of U-Col4 or urinary albumin excreted. The decline in estimated glomerular filtration rate (eGFR) was calculated. In participants with eGFR ≥80 mL/min, abnormal U-Col4 excretion was indicated as a significant independent risk factor for 10% eGFR change per year, which is one of the prognostic factors for the development of end-stage kidney disease. Moreover, in contrast to urinary albumin excretion, U-Col4 excretion was not related to age or kidney function, suggesting that some individuals with abnormal U-Col4 excretion can have an independent hidden risk for the development of kidney dysfunction. In conclusion, it is important to measure U-Col4 excretion in the general population without diabetes to determine changes in renal features in every individual and help detect future complications such as diabetic kidney disease. If U-Col4 excretion is abnormal, kidney manifestation should be carefully followed up, even if the kidney function and urinalysis findings are normal.
(キーワード)
Adult / Aged / Aged, 80 and over / Albuminuria / Asian Continental Ancestry Group / Collagen Type IV / Diabetic Nephropathies / Disease Progression / Female / Glomerular Filtration Rate / Humans / Japan / Kaplan-Meier Estimate / Male / Middle Aged / Prognosis / Renal Insufficiency / Risk Factors / Young Adult
Toshio Doi, Hideharu Abe, Seiji Kishi, Taichi Murakami, Kojiro Nagai and Tatsuya Tominaga : Urinary IgG4 and Smad1 are Specific Biomarkers for Renal Structural and Functional Changes in Early Stages of Diabetic Nephropathy., Diabetes, Vol.67, No.5, 986-993, 2018.
(要約)
Diabetic nephropathy (DN) is the major cause of end-stage kidney disease, but early biomarkers of DN risk are limited. Herein we examine urinary IgG4 and Smad1 as additional early DN biomarkers. We recruited 815 patients with type 2 diabetes; 554 patients fulfilled the criteria of an estimated glomerular filtration rate (eGFR) >60 mL/min and no macroalbuminuria at baseline, with follow-up for 5 years. Patients without macroalbuminuria were also recruited for renal biopsies. Urinary IgG4 and Smad1 were determined by enzyme-linked immunoassays using specific antibodies. The specificity, sensitivity, and reproducibility were confirmed for each assay. Increased urinary IgG4 was significantly associated with lower eGFR. The level of urinary IgG4 also significantly correlated with surface density of peripheral glomerular basement membrane (Sv PGBM/Glom), whereas Smad1 was associated with the degree of mesangial expansion-both classic pathological findings in DN. Baseline eGFR did not differ between any groups; however, increases in both urinary IgG4 and Smad1 levels at baseline significantly predicted later development of eGFR decline in patients without macroalbuminuria. These data suggest that urinary IgG4 and Smad1 at relatively early stages of DN reflect underlying DN lesions and are relevant to later clinical outcomes.
(キーワード)
Adult / Biomarkers / Diabetes Mellitus, Type 2 / Diabetic Nephropathies / Early Diagnosis / Enzyme-Linked Immunosorbent Assay / Female / Glomerular Basement Membrane / Glomerular Filtration Rate / Humans / Immunoglobulin G / Kidney / Male / Mesangial Cells / Microscopy, Electron / Middle Aged / Reproducibility of Results / Sensitivity and Specificity / Smad1 Protein
Hideharu Abe and Tatsuya Tominaga : New insights into the molecular pathology and the development of predictive biomarkers in diabetic mephropathy., Trends in Cell & Molecular Biology, Vol.13, No.4, 19-26, 2018.
11.
Kojiro Nagai, Tatsuya Tominaga, Sayo Ueda, Eriko Shibata, Masanori Tamaki, Motokazu Matsuura, Seiji Kishi, Taichi Murakami, Tatsumi Moriya, Hideharu Abe and Toshio Doi : Mesangial Cell Mammalian Target of Rapamycin Complex 1 Activation Results in Mesangial Expansion., Journal of the American Society of Nephrology, Vol.28, No.10, 2879-2885, 2017.
(要約)
Human glomerular diseases can be caused by several different diseases, many of which include mesangial expansion and/or proliferation followed by glomerulosclerosis. However, molecular mechanisms underlying the pathologic mesangial changes remain poorly understood. Here, we investigated the role of the mammalian target of rapamycin complex 1 (mTORC1)-S6 kinase pathway in mesangial expansion and/or proliferation by ablating an upstream negative regulator, tuberous sclerosis complex 1 (TSC1), using tamoxifen-induced Foxd1-Cre mice [Foxd1ER(+) TSC1 mice]. Foxd1ER(+) TSC1 mice showed mesangial expansion with increased production of collagen IV, collagen I, and -smooth muscle actin in glomeruli, but did not exhibit significant mesangial proliferation or albuminuria. Furthermore, rapamycin treatment of Foxd1ER(+) TSC1 mice suppressed mesangial expansion. Among biopsy specimens from patients with glomerular diseases, analysis of phosphorylated ribosomal protein S6 revealed mesangial cell mTORC1 activation in IgA nephropathy and in lupus mesangial proliferative nephritis but not in the early phase of diabetic nephropathy. In summary, mesangial cell mTORC1 activation can cause mesangial expansion and has clinical relevance for human glomerular diseases. This report also confirms that the tamoxifen-induced mesangium-specific Cre-loxP system is useful for studies designed to clarify the role of the mesangium in glomerular diseases in adults.
Motokazu Matsuura, Hideharu Abe, Tatsuya Tominaga, Akiko Sakurai, Taichi Murakami, Seiji Kishi, Yoshimi Bando, Jun Minakuchi, Kojiro Nagai and Toshio Doi : A Novel Method of DAPI Staining for Differential Diagnosis of Renal Amyloidosis., The Journal of Medical Investigation : JMI, Vol.64, No.3.4, 217-221, 2017.
(要約)
Amyloidosis is often overlooked because its clinical manifestations can mimic those of more-common diseases. It is important to get a precise diagnosis as early as possible for the prevention of further organ damages. Amyloidosis is a disorder caused by deposition of insoluble abnormal amyloid. The kidney is a frequent site of amyloid deposition. The amyloid fibrils have a characteristic appearance and generate birefringence under polarized light when stained with the Congo red dye. Classification of amyloidosis is based on the precursor protein that forms the amyloid fibrils and the distribution of amyloid deposits as either systemic or localized. Involvement of amyloid fibrils in kidneys mainly occurs as amyloid light-chain (AL) or amyloid A (AA) amyloidosis. The potassium permanganate method with Congo red staining was once used widely to discriminate AL and AA amyloidoses, but this method has a problem of false positive results. We found that extracellular and cytoplasmic glomerular 4', 6-diamidino-2-phenylindole (DAPI)-positive areas were clearly consistent with amyloid deposition in AL amyloidosis. In contrast, the overlapping staining was not seen in AA amyloidosis. Therefore, we propose that DAPI staining readily distinguishes AL renal amyloidosis from AA renal amyloidosis as a simple and reproducible histochemical method. J. Med. Invest. 64: 217-221, August, 2017.
(キーワード)
Amyloidosis / Kidney Diseases / Serum Amyloid A Protein / Staining and Labeling
Tatsuya Tominaga, K Rajesh Dutta, Darukeshwara Joladarashi, Toshio Doi, K Janardan Reddy and S Yashpal Kanwar : Transcriptional and Translational Modulation of myo-Inositol Oxygenase (Miox) by Fatty Acids: IMPLICATIONS IN RENAL TUBULAR INJURY INDUCED IN OBESITY AND DIABETES., The Journal of Biological Chemistry, Vol.291, No.3, 1348-1367, 2015.
(要約)
The kidney is one of the target organs for various metabolic diseases, including diabetes, metabolic syndrome, and obesity. Most of the metabolic studies underscore glomerular pathobiology, although the tubulo-interstitial compartment has been underemphasized. This study highlights mechanisms concerning the pathobiology of tubular injury in the context of myo-inositol oxygenase (Miox), a tubular enzyme. The kidneys of mice fed a high fat diet (HFD) had increased Miox expression and activity, and the latter was related to phosphorylation of serine/threonine residues. Also, expression of sterol regulatory element-binding protein1 (Srebp1) and markers of cellular/nuclear damage was increased along with accentuated apoptosis and loss of tubular brush border. Similar results were observed in cells treated with palmitate/BSA. Multiple sterol-response elements and E-box motifs were found in the miox promoter, and its activity was modulated by palmitate/BSA. Electrophoretic mobility and ChIP assays confirmed binding of Srebp to consensus sequences of the miox promoter. Exposure of palmitate/BSA-treated cells to rapamycin normalized Miox expression and prevented Srebp1 nuclear translocation. In addition, rapamycin treatment reduced p53 expression and apoptosis. Like rapamycin, srebp siRNA reduced Miox expression. Increased expression of Miox was associated with the generation of reactive oxygen species (ROS) in kidney tubules of mice fed an HFD and cell exposed to palmitate/BSA. Both miox and srebp1 siRNAs reduced generation of ROS. Collectively, these findings suggest that HFD or fatty acids modulate transcriptional, translational, and post-translational regulation of Miox expression/activity and underscore Miox being a novel target of the transcription factor Srebp1. Conceivably, activation of the mTORC1/Srebp1/Miox pathway leads to the generation of ROS culminating into tubulo-interstitial injury in states of obesity.
Takahiro Hirano, Taichi Murakami, Hiroyuki Ono, Akiko Sakurai, Tatsuya Tominaga, Toshikazu Takahashi, Kojiro Nagai, Toshio Doi and Hideharu Abe : A Novel Interaction between FLICE-Associated Huge Protein (FLASH) and E2A Regulates Cell Proliferation and Cellular Senescence via Tumor Necrosis Factor (TNF)-Alpha-p21WAF1/CIP1 Axis., PLoS ONE, Vol.10, No.7, e0133205, 2015.
(要約)
Dysregulation of the cell proliferation has been implicated in the pathophysiology of a number of diseases. Cellular senescence limits proliferation of cancer cells, preventing tumorigenesis and restricting tissue damage. However, the role of cellular senescence in proliferative nephritis has not been determined. The proliferative peak in experimental rat nephritis coincided with a peak in E2A expression in the glomeruli. Meanwhile, E12 (an E2A-encoded transcription factor) did not promote proliferation of Mesangial cells (MCs) by itself. We identified caspase-8-binding protein FLICE-associated huge protein (FLASH) as a novel E2A-binding partner by using a yeast two-hybrid screening. Knockdown of FLASH suppressed proliferation of MCs. This inhibitory effect was partially reversed by the knockdown of E2A. In addition, the knockdown of FLASH induced cyclin-dependent kinase inhibitor p21WAF1/CIP1 (p21) expression, but did not affect p53 expression. Furthermore, overexpression of E12 and E47 induced p21, but not p53 in MCs, in the absence of FLASH. We also demonstrated that E2A and p21 expression at the peak of proliferation was followed by significant induction of FLASH in mesangial areas in rat proliferative glomerulonephritis. Moreover, we revealed that FLASH negatively regulates cellular senescence via the interaction with E12. We also demonstrated that FLASH is involved in the TNF--induced p21 expressions. These results suggest that the functional interaction of E2A and FLASH play an important role in cell proliferation and cellular senescence via regulation of p21 expression in experimental glomerulonephritis.
Takeshi Matsubara, Makoto Araki, Hideharu Abe, Otoya Ueda, Kou-Ichi Jishage, Akira Mima, Chisato Goto, Tatsuya Tominaga, Masahiko Kinosaki, Seiji Kishi, Kojiro Nagai, Noriyuki Iehara, Naoshi Fukushima, Toru Kita, Hidenori Arai and Toshio Doi : Bone Morphogenetic Protein 4 and Smad1 Mediate Extracellular Matrix Production in the Development of Diabetic Nephropathy., Diabetes, Vol.64, No.8, 2978-2990, 2015.
(要約)
Diabetic nephropathy is the leading cause of end-stage renal disease. It is pathologically characterized by the accumulation of extracellular matrix in the mesangium, of which the main component is 1/2 type IV collagen (Col4a1/a2). Recently, we identified Smad1 as a direct regulator of Col4a1/a2 under diabetic conditions in vitro. Here, we demonstrate that Smad1 plays a key role in diabetic nephropathy through bone morphogenetic protein 4 (BMP4) in vivo. Smad1-overexpressing mice (Smad1-Tg) were established, and diabetes was induced by streptozotocin. Nondiabetic Smad1-Tg did not exhibit histological changes in the kidney; however, the induction of diabetes resulted in an 1.5-fold greater mesangial expansion, consistent with an increase in glomerular phosphorylated Smad1. To address regulatory factors of Smad1, we determined that BMP4 and its receptor are increased in diabetic glomeruli and that diabetic Smad1-Tg and wild-type mice treated with a BMP4-neutralizing antibody exhibit decreased Smad1 phosphorylation and 40% less mesangial expansion than those treated with control IgG. Furthermore, heterozygous Smad1 knockout mice exhibit attenuated mesangial expansion in the diabetic condition. The data indicate that BMP4/Smad1 signaling is a critical cascade for the progression of mesangial expansion and that blocking this signal could be a novel therapeutic strategy for diabetic nephropathy.
Kazuhiro Yoshikawa, Hideharu Abe, Tatsuya Tominaga, Masayuki Nakamura, Seiji Kishi, Motokazu Matsuura, Kojiro Nagai, Kenji Tsuchida, Jun Minakuchi and Toshio Doi : Polymorphism in the human matrix Gla protein gene is associated with the progression of vascular calcification in maintenance hemodialysis patients, Clinical and Experimental Nephrology, Vol.17, No.6, 882-889, 2013.
(要約)
Matrix Gla protein (MGP) is one of the important proteins inhibiting vascular calcification (VC). Single nucleotide polymorphisms (SNPs) located in the promoter and coding regions of the MGP gene affect the transcriptional activity. In this study, we investigated the relationship between the SNPs and progression of VC in patients undergoing maintenance hemodialysis (MHD). This was a retrospective, longitudinal cohort study of 134 MHD patients whose VC could be followed by multi-detector computed tomography (MDCT) examinations. MGP-SNPs (T-138C, rs1800802 and G-7A, rs1800801) were determined. The progression speed of VC was examined by plotting the abdominal aortic calcium volume scores. The progression speed of VC of patients with the CC genotype of T-138C was significantly slower than that of patients with the CT or TT genotype. Multiple regression analysis showed that CT/TT genotype, greater age at the beginning of MHD, male sex, high levels of calcium × phosphate, low levels of high-density lipoprotein cholesterol, high levels of low-density lipoprotein cholesterol, low levels of ferritin and non-use of angiotensin II receptor blockers were significantly associated with progression of VC. The MGP-138CC genotype may be associated with slower progression of VC in MHD patients. The genotype of the MGP gene will be a genomic biomarker that is predictive of VC progression.
Kojiro Nagai, Masashi Miyoshi, Takei Kake, Naoshi Fukushima, Motokazu Matsuura, Eriko Shibata, Satoshi Yamada, Kazuhiro Yoshikawa, Hiro-omi Kanayama, Tomoya Fukawa, Kunihisa Yamaguchi, Hirofumi Izaki, Akira Mima, Naoko Abe, Toshikazu Araoka, Taichi Murakami, Fumi Kishi, Seiji Kishi, Tatsuya Tominaga, Tatsumi Moriya, Hideharu Abe and Toshio Doi : Dual Involvement of Growth Arrest-specific Gene 6 in the Early Phase of Human IgA Nephropathy, PLoS ONE, Vol.8, No.6, e66759, 2013.
(要約)
Gas6 is a growth factor that causes proliferation of mesangial cells in the development of glomerulonephritis. Gas6 can bind to three kinds of receptors; Axl, Dtk, and Mer. However, their expression and functions are not entirely clear in the different glomerular cell types. Meanwhile, representative cell cycle regulatory protein p27 has been reported to be expressed in podocytes in normal glomeruli with decreased expression in proliferating glomeruli, which inversely correlated with mesangial proliferation in human IgA nephropathy (IgAN). The aim of this study is to clarify Gas6 involvement in the progression of IgAN. Expression of Gas6/Axl/Dtk was examined in 31 biopsy proven IgAN cases. We compared the expression levels with histological severity or clinical data. Moreover, we investigated the expression of Gas6 and its receptors in cultured podocytes. In 28 of 31 cases, Gas6 was upregulated mainly in podocytes. In the other 3 cases, Gas6 expression was induced in endothelial and mesangial cells, which was similar to animal nephritis models. Among 28 podocyte type cases, the expression level of Gas6 correlated with the mesangial hypercellularity score of IgAN Oxford classification and urine protein excretion. It also inversely correlated with p27 expression in glomeruli. As for the receptors, Axl was mainly expressed in endothelial and mesangial cells, while Dtk was expressed in podocytes. In vitro, Dtk was expressed in cultured murine podocytes, and the expression of p27 was decreased by Gas6 stimulation. Gas6 was uniquely upregulated in either endothelial/mesangial cells or podocytes in IgAN. The expression pattern can be used as a marker to classify IgAN. Gas6 has a possibility to be involved in not only mesangial proliferation via Axl, but also podocyte injury via Dtk in IgAN.
Hideharu Abe, Tatsuya Tominaga, Takeshi Matsubara, Naoko Abe, Seiji Kishi, Kojiro Nagai, Taichi Murakami, Toshikazu Araoka and Toshio Doi : Scleraxis modulates bone morphogenetic protein 4 (BMP4)-Smad1 protein-smooth muscle -actin (SMA) signal transduction in diabetic nephropathy., The Journal of Biological Chemistry, Vol.287, No.24, 20430-20442, 2012.
(要約)
Activation of mesangial cells (MCs), which is characterized by induction of smooth muscle -actin (SMA) expression, contributes to a key event in various renal diseases; however, the mechanisms controlling MC differentiation are still largely undefined. Activated Smad1 induced SMA in a dose-dependent manner in MCs. As a direct regulating molecule for SMA, we identified and characterized scleraxis (Scx) as a new phenotype modulator in advanced glycation end product (AGE)-exposed MCs. Scx physically associated with E12 and bound the E-box in the promoter of SMA and negatively regulated the AGE-induced SMA expression. Scx induced expression and secretion of bone morphogenetic protein 4 (BMP4), thereby controlling the Smad1 activation in AGE-treated MCs. In diabetic mice, Scx was concomitantly expressed with SMA in the glomeruli. Inhibitor of differentiation 1 (Id1) was further induced by extended treatment with AGE, thereby dislodging Scx from the SMA promoter. These data suggest that Scx and Id1 are involved in the BMP4-Smad1-SMA signal transduction pathway besides the TGF1-Smad1-SMA signaling pathway and modulate phenotypic changes in MCs in diabetic nephropathy.
Seiji Kishi, Hideharu Abe, Haruhiko Akiyama, Tatsuya Tominaga, Taichi Murakami, Akira Mima, Kojiro Nagai, Fumi Kishi, Motokazu Matsuura, Takeshi Matsubara, Noriyuki Iehara, Otoya Ueda, Naoshi Fukushima, Kou-Ichi Jishage and Toshio Doi : SOX9 protein induces a chondrogenic phenotype of mesangial cells and contributes to advanced diabetic nephropathy., The Journal of Biological Chemistry, Vol.286, No.37, 32162-32169, 2011.
(要約)
Diabetic nephropathy (DN) is the most important chronic kidney disease. We previously reported that Smad1 transcriptionally regulates the expression of extracellular matrix in DN. Phenotypic change in mesangial cells (MCs) is a key pathologic event in the progression of DN. The aim of this study is to investigate a novel mechanism underlying chondrogenic phenotypic change in MCs that results in the development of DN. MCs showed chondrogenic potential in a micromass culture, and BMP4 induced the expression of chondrocyte markers (SRY-related HMG Box 9 (SOX9) and type II collagen (COL2)). Advanced glycation end products induced the expression of chondrocyte marker proteins downstream from the BMP4-Smad1 signaling pathway in MCs. In addition, hypoxia also induced the expression of BMP4, hypoxia-inducible factor-1 (HIF-1), and chondrocyte markers. Overexpression of SOX9 caused ectopic expression of proteoglycans and COL2 in MCs. Furthermore, forced expression of Smad1 induced chondrocyte markers as well. Dorsomorphin inhibited these inductions. Glomerular expressions of HIF-1, BMP4, and chondrocyte markers were observed in diabetic nephropathy mice. These positive stainings were observed in mesangial sclerotic lesions. SOX9 was partially colocalized with HIF-1 and BMP4 in diabetic glomeruli. BMP4 knock-in transgenic mice showed not only similar pathological lesions to DN, but also the induction of chondrocyte markers in the sclerotic lesions. Here we demonstrate that HIF-1 and BMP4 induce SOX9 expression and subsequent chondrogenic phenotype change in DN. The results suggested that the transdifferentiation of MCs into chondrocyte-like cells in chronic hypoxic stress may result in irreversible structural change in DN.
(キーワード)
Animals / Antigens, Differentiation / Bone Morphogenetic Protein 4 / Cell Line / Cell Transdifferentiation / Chondrocytes / Collagen Type II / Diabetic Nephropathies / Gene Expression Regulation / Glomerular Mesangium / Glycosylation End Products, Advanced / Hypoxia-Inducible Factor 1, alpha Subunit / Mice / Mice, Transgenic / SOX9 Transcription Factor / Smad1 Protein
Tatsuya Tominaga, Hideharu Abe, Otoya Ueda, Chisato Goto, Kunihiko Nakahara, Taichi Murakami, Takeshi Matsubara, Akira Mima, Kojiro Nagai, Toshikazu Araoka, Seiji Kishi, Naoshi Fukushima, Kou-ichi Jishage and Toshio Doi : Activation of bone morphogenetic protein 4 signaling leads to glomerulosclerosis that mimics diabetic nephropathy., The Journal of Biological Chemistry, Vol.286, No.22, 20109-20116, 2011.
(要約)
Diabetic nephropathy (DN) is the most common cause of chronic kidney disease. We have previously reported that Smad1 transcriptionally regulates the expression of extracellular matrix (ECM) proteins in DN. However, little is known about the regulatory mechanisms that induce and activate Smad1. Here, bone morphogenetic protein 4 (Bmp4) was found to up-regulate the expression of Smad1 in mesangial cells and subsequently to phosphorylate Smad1 downstream of the advanced glycation end product-receptor for advanced glycation end product signaling pathway. Moreover, Bmp4 utilized Alk3 and affected the activation of Smad1 and Col4 expressions in mesangial cells. In the diabetic mouse, Bmp4 was remarkably activated in the glomeruli, and the mesangial area was expanded. To elucidate the direct function of Bmp4 action in the kidneys, we generated transgenic mice inducible for the expression of Bmp4. Tamoxifen treatment dramatically induced the expression of Bmp4, especially in the glomeruli of the mice. Notably, in the nondiabetic condition, the mice exhibited not only an expansion of the mesangial area and thickening of the basement membrane but also remarkable albuminuria, which are consistent with the distinct glomerular injuries in DN. ECM protein overexpression and activation of Smad1 in the glomeruli were also observed in the mice. The mesangial expansion in the mice was significantly correlated with albuminuria. Furthermore, the heterozygous Bmp4 knock-out mice inhibited the glomerular injuries compared with wild type mice in diabetic conditions. Here, we show that BMP4 may act as an upstream regulatory molecule for the process of ECM accumulation in DN and thereby reveals a new aspect of the molecular mechanisms involved in DN.
(キーワード)
Albuminuria / Animals / Bone Morphogenetic Protein 4 / Diabetic Nephropathies / 細胞外マトリックス (extracellular matrix) / Glomerulosclerosis, Focal Segmental / Glycosylation End Products, Advanced / Mesangial Cells / Mice / ノックアウトマウス (knockout mice) / Smad1 Protein / Up-Regulation
Akira Mima, Hideharu Abe, Kojiro Nagai, Hidenori Arai, Takeshi Matsubara, Makoto Araki, Kazuo Torikoshi, Tatsuya Tominaga, Noriyuki Iehara, Atsushi Fukatsu, Toru Kita and Toshio Doi : Activation of Src mediates PDGF-induced Smad1 phosphorylation and contributes to the progression of glomerulosclerosis in glomerulonephritis., PLoS ONE, Vol.6, No.3, e17929, 2011.
(要約)
Platelet-derived growth factor (PDGF) plays critical roles in mesangial cell (MC) proliferation in mesangial proliferative glomerulonephritis. We showed previously that Smad1 contributes to PDGF-dependent proliferation of MCs, but the mechanism by which Smad1 is activated by PDGF is not precisely known. Here we examined the role of c-Src tyrosine kinase in the proliferative change of MCs. Experimental mesangial proliferative glomerulonephritis (Thy1 GN) was induced by a single intravenous injection of anti-rat Thy-1.1 monoclonal antibody. In Thy1 GN, MC proliferation and type IV collagen (Col4) expression peaked on day 6. Immunohistochemical staining for the expression of phospho-Src (pSrc), phospho-Smad1 (pSmad1), Col4, and smooth muscle -actin (SMA) revealed that the activation of c-Src and Smad1 signals in glomeruli peaked on day 6, consistent with the peak of mesangial proliferation. When treated with PP2, a Src inhibitor, both mesangial proliferation and sclerosis were significantly reduced. PP2 administration also significantly reduced pSmad1, Col4, and SMA expression. PDGF induced Col4 synthesis in association with increased expression of pSrc and pSmad1 in cultured MCs. In addition, PP2 reduced Col4 synthesis along with decreased pSrc and pSmad1 protein expression in vitro. Moreover, the addition of siRNA against c-Src significantly reduced the phosphorylation of Smad1 and the overproduction of Col4. These results provide new evidence that the activation of Src/Smad1 signaling pathway plays a key role in the development of glomerulosclerosis in experimental glomerulonephritis.
Taichi Murakami, Hideharu Abe, Kojiro Nagai, Tatsuya Tominaga, Norimichi Takamatsu, Toshikazu Araoka, Seiji Kishi, Toshikazu Takahashi, Akira Mima, Yoshimi Takai, Jeffrey B. Kopp and Toshio Doi : Trophoblast glycoprotein: possible candidate mediating podocyte injuries in glomerulonephritis., American Journal of Nephrology, Vol.32, No.6, 505-521, 2010.
(要約)
trophoblast glycoprotein (Tpbg), a 72-kDa transmembrane glycoprotein, is known to regulate the phenotypes of epithelial cells by modifying actin organization and cell motility. Recently, a microarray study showed that Tpbg is upregulated in Thy1 glomerulonephritis (Thy1 GN). We hypothesized that Tpbg regulates cytoskeletal rearrangement and modulates phenotypic alteration in podocytes under pathological conditions.
Norimichi Takamatsu, Hideharu Abe, Tatsuya Tominaga, Kunihiko Nakahara, Yumi Ito, Yoko Okumoto, Jiyoong Kim, Masafumi Kitakaze and Toshio Doi : Risk factors for chronic kidney disease in Japan: a community-based study., BMC Nephrology, Vol.10, 34, 2009.
(要約)
Chronic kidney disease (CKD) is increasingly being recognized as a predictor for both end-stage renal disease and cardiovascular disease. The present study, conducted on individuals from a community in Arita, Japan, was designed to evaluate biomarkers that can be used to determine the associated factors for CKD.
(キーワード)
Adult / Aged / Aged, 80 and over / Cross-Sectional Studies / Female / Glomerular Filtration Rate / Humans / 日本 (Japan) / Kidney Failure, Chronic / Male / Middle Aged / Residence Characteristics / Risk Factors / Young Adult
Toshio Doi, Akira Mima, Takeshi Matsubara, Tatsuya Tominaga, Hidenori Arai and Hideharu Abe : The current clinical problems for early phase of diabetic nephropathy and approach for pathogenesis of diabetic nephropathy., Diabetes Research and Clinical Practice, Vol.82, No.Suppl 1, S21-4, 2008.
(要約)
The important clinical problems of diabetic nephropathy are both proteinuria and decrease of renal function. Pathological analysis showed decrease of GFR was correlated to degree of mesangial expansion but not thickening of GBM nor the other findings in human type 1 diabetic nephropathy. From the perspective in renal dysfunction, mesangial matrix expansion was crucial for diabetic nephropathy. However, there was no difference of mesangial expansion between normal and microalbuminuria stage in type 1 and 2 diabetes mellitus (DM). On the other hand, microalbuminuria definitely shows a key related factor for cardiovascular events, but it does not indicate a clear interaction for glomerulosclerosis. We need to search a new clinical marker for renal injury. We have first shown that Smad1 is a transcription factor for alpha1 and 2 of type 4 collagen (Col4), which is a major component of glomerulosclerosis. We have also identified Smad1 is a critical responsible molecule for developing glomerulosclerosis in rat diabetic nephropathy. We have found the good correlation between glomerulosclerosis and urinary Smad1 but not between glomerulosclerosis and urine albumin. These data suggests that urine Smad1 is a promising clinical marker for underlying glomerular damages in early stage diabetic nephropathy. The study also implicates that angiotensin II (AngII)-Src-Smad1 signaling pathway has played a key role for development of diabetic nephropathy. These suggest that it is necessary to clarify the whole mechanism related to Smad1 to identify the pathogenesis of diabetic nephropathy.
Akira Mima, Hidenori Arai, Takeshi Matsubara, Hideharu Abe, Kojiro Nagai, Yukinori Tamura, Kazuo Torikoshi, Makoto Araki, Hiroshi Kanamori, Toshikazu Takahashi, Tatsuya Tominaga, Motokazu Matsuura, Noriyuki Iehara, Atsushi Fukatsu, Toru Kita and Toshio Doi : Urinary Smad1 is a novel marker to predict later onset of mesangial matrix expansion in diabetic nephropathy., Diabetes, Vol.57, No.6, 1712-1722, 2008.
(要約)
We reported that Smad1 is a key transcriptional factor for mesangial matrix expansion in diabetic nephropathy. In this study, we examined whether urinary Smad1 in an early phase of diabetes can predict later development of glomerulosclerosis in diabetic nephropathy and how an angiotensin II type 1 receptor blocker (ARB) can modulate structural changes and urinary markers.
Masanori Tamaki, Tatsuya Tominaga, Yui Fujita, Taichi Murakami, Seiji Kishi, Hideharu Abe, Kojiro Nagai and Toshio Doi : A Novel All-Trans Retinoic Acid Therapy Directly Suppresses Bone Morphogenetic Protein 4 in Mouse Diabetic Nephropathy, KIDNEY WEEK 2019 American Society of Nephrology, Nov. 2019.
2.
Masanori Tamaki, Tatsuya Tominaga, Yui Fujita, Seiji Kishi, Taichi Murakami, Kojiro Nagai, Hideharu Abe and Toshio Doi : Mesangial Matrix Expansion Attenuated by All-Trans Retinoic Acid Through Direct Suppression of Bone Morphogenetic Protein 4 in Mouse Diabetic Nephropathy, KIDNEY WEEK 2018 American Society of Nephrology, Oct. 2018.
3.
Yui Fujita, Tatsuya Tominaga, Masanori Tamaki, Taichi Murakami, Seiji Kishi, Kojiro Nagai, Hideharu Abe and Toshio Doi : Palovarotene, Selective Retinoic Receptor- Agonist, Inhibited Both BMP4 and TGF- Signaling Pathways in Diabetic Nephropathy, KIDNEY WEEK 2017 American Society of Nephrology, Nov. 2017.
4.
Masanori Tamaki, Tatsuya Tominaga, Yui Fujita, Seiji Kishi, Taichi Murakami, Kojiro Nagai, Hideharu Abe and Toshio Doi : Glomerulosclerosis Attenuated by Retinoic Acid through Bone Morphogenetic Protein 4 Suppression in Mice with Streptozotocin-Induced Diabetes, KIDNEY WEEK 2017 American Society of Nephrology, Nov. 2017.
5.
Kojiro Nagai, Tatsuya Tominaga, Taichi Murakami, Eriko Shibata, Seiji Kishi, Motokazu Matsuura, Hideharu Abe and Toshio Doi : In vivo evidence of mTORC1-S6 kinase pathway involvement in the development of glomerulosclerosis in human IgA nephropathy, KIDNEY WEEK 2016 American Society of Nephrology, Nov. 2016.
6.
Yui Fujita, Tatsuya Tominaga, Taichi Murakami, Seiji Kishi, Kojiro Nagai, Hideharu Abe and Toshio Doi : The Role of BMP4 Signal Pathway on the Podocyte Injury in Diabetic Early Stage, ASN Kidney week 2016, Nov. 2016.
7.
Tatsuya Tominaga, Toshio Doi and Kanwar S Yashpal : Myo-inositol oxygenase (MIOX) contributes to renal tubular injury through ER stress and a hyaluronic acid (HA) production, American Society of Nephrology Kidney Week 2015, Nov. 2015.
8.
Tupe Santosh Rashmi, Tatsuya Tominaga and Kanwar S. Yashpal : Advanced Glycation End Products Induces Myo-Inositol Oxygenase via Activation of PI3K Pathwayin Proximal Tubular Epithelial Cells, American Society of Nephrology Kidney Week 2014, Nov. 2014.
9.
Zhan Ming, Tatsuya Tominaga, Sun Lin and Kanwar S Yashpal : Myo-Inositol Oxygenase (MIOX) Modulates Mitochondrial Dynamics and Autophagy in Diabetic Tubulopathy, American Society of Nephrology Kidney Week 2013, Nov. 2013.
10.
Tatsuya Tominaga, Zhan Ming and Kanwar S Yashpal : Relevance of Myo-Inositol Oxigenase (MIOX) in Renal Tubular Injury in Obesity/Metabolic Syndrome, American Society of Nephrology Kidney Week 2013, Nov. 2013.
11.
Tatsuya Tominaga, Hideharu Abe, Kojiro Nagai, Akira Mima, Seiji Kishi, K Jishage, N Fukushima and Toshio Doi : Role of BMP4 for regulating podocyte injury in the diabetic nephropathy., American Society for Cell biology, San Francisco, Dec. 2012.
12.
Masashi Miyoshi, Kojiro Nagai, Takei Kake, Naoshi Fukushima, Motokazu Matsuura, Eriko Shibata, Satoshi Yamada, Kazuhiro Yoshikawa, Akira Mima, Fumi Kishi, Seiji Kishi, Tatsuya Tominaga, Hideharu Abe and Toshio Doi : Expression of Gas6 and Axl in Human IgA Nephropathy. ~A Possible Involvement of Gas6 in Podocyte Injury~, American Society of Nephrology Kidney Week 2012, San Diego, Nov. 2012.
13.
Tatsuya Tominaga, Hideharu Abe, Kojiro Nagai, Seiji Kishi, Akira Mima and Toshio Doi : BMP4 regulates podocyte injury in the diabetic nephropathy, Diabetes, Philadelphia, May 2012.
14.
Tatsuya Tominaga, Hideharu Abe, Kojiro Nagai, Seiji Kishi and Toshio Doi : BMP4 regulates podocyte injury in the diabetic nephropathy, The 1st Asia-Pacific Vascular Biology Meeting, Dec. 2011.
15.
Tatsuya Tominaga, Hideharu Abe, Kojiro Nagai, Seiji Kishi, Taichi Murakami and Toshio Doi : BMP4 regulates podocyte injury in the diabetic nephropathy, Journal of the American Society of Nephrology, Nov. 2011.
16.
Akira Mima, H Arai, T Matsubara, Hideharu Abe, H Kanamori, E Sumi, Tatsuya Tominaga, T Takahashi, Motokazu Matsuura, N Iehara, A Fukatsu, T Kita and Toshio Doi : Angiotensin II type I receptor blocker decreases urinary Smad1, a marker for mesangial matrix expansion in diabetic nephropathy, 2007 World Congress of Nephrology, リオデジャネイロ, Apr. 2007.