Screening of novel podocyte protective reagents using nephrin reporter mice

• Project/Area Number: 16K15469
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Kidney internal medicine
• Research Institution: University of Shizuoka
• Principal Investigator: Mori Kiyoshi 静岡県立大学, 薬学部, 特任教授 (60343232)
• Co-Investigator(Kenkyū-buntansha): 松阪 泰二 東海大学, 付置研究所, 助教授 (50317749)
• Co-Investigator(Renkei-kenkyūsha): OHTSUKA Masato 東海大学, 医学部, 准教授 (90372945)
• Research Collaborator: TSUCHIDA Junichi 京都大学, 医学部, 研究員 (10643570)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 腎臓内科学 / 糸球体 / ポドサイト / 化合物ライブラリー / スクリーニング / レポーターアッセイ / 腎臓学 / ネフリン / レポーター

Outline of Final Research Achievements

Nephrin is a critical component of glomerular filtration barrier. Downregulation of nephrin expression is commonly observed at early stage of glomerular disorders, suggesting that methods to increase nephrin expression in podocytes may have therapeutic utility. Here, we generated a knockin mouse line carrying single copy of 5.5 kb nephrin promoter controlling expression of enhanced green fluorescent protein (EGFP) at Rosa26 genomic locus (Nephrin-EGFP mouse). Next, we isolated and cultivated glomeruli from these mice, and developed a protocol to automatically quantitate EGFP expression in cultured glomeruli. EGFP signal was markedly reduced after 5 days of culture but reduction was inhibited by vitamin D treatment. Thus, we generated a mouse line converting nephrin promoter activity into fluorescence, which can be used to screen compounds having activity to enhance nephrin gene expression.

Research Products

• [Journal Article] Increase of Total Nephron Albumin Filtration and Reabsorption in Diabetic Nephropathy. (2017)
  • Author(s): Mori KP, Yokoi H, Kasahara M, Imamaki H, Ishii A, Kuwabara T, Koga K, Kato Y, Toda N, Ohno S, Kuwahara K, Endo T, Nakao K, Yanagita M, Mukoyama M, Mori K.
  • Journal Title: J Am Soc Nephrol. Volume: 28 Issue: 1 Pages: 278-289
  • DOI: 10.1681/asn.2015101168
  • NAID: 120005820888
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Obesity-promoting and anti-thermogenic effects of neutrophil gelatinase-associated lipocalin in mice (2017)
  • Author(s): Ishii Akira、Katsuura Goro、Imamaki Hirotaka、Kimura Hiroyuki、Mori Keita P.、Kuwabara Takashige、Kasahara Masato、Yokoi Hideki、Ohinata Kousaku、Kawanishi Tomoko、Tsuchida Junichi、Nakamoto Yuji、Nakao Kazuwa、Yanagita Motoko、Mukoyama Masashi、Mori Kiyoshi
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 15501-15501
  • DOI: 10.1038/s41598-017-15825-4
  • Peer Reviewed / Open Access
• [Journal Article] Crucial role of mesangial cell-derived connective tissue growth factor in a mouse model of anti-glomerular basement membrane glomerulonephritis. (2017)
  • Author(s): Toda N, Mori K, Kasahara M, Ishii A, Koga K, Ohno S, Mori KP. Kato Y, Osaki K, Kuwabara T, Kojima K, Taura D, Sone M, Matsusaka T, Nakao K, Mukoyama M, Yanagita M, Yokoi H.
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 42114-42114
  • DOI: 10.1038/srep42114
  • NAID: 120005997757
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Glomerulosclerosis Induced by Deficiency of Membrane-Associated Guanylate Kinase Inverted 2 in Kidney Podocytes (2017)
  • Author(s): Shirata Naritoshi、Ihara Kan-ichiro、Yamamoto-Nonaka Kanae、Seki Takuto、Makino Shin-ichi、Oliva Trejo Juan Alejandro、Miyake Takafumi、Yamada Hiroyuki、Campbell Kirk Nicholas、Nakagawa Takahiko、Mori Kiyoshi、Yanagita Motoko、Mundel Peter、Nishimori Katsuhiko、Asanuma Katsuhiko
  • Journal Title: Journal of the American Society of Nephrology Volume: 28 Issue: 9 Pages: 2654-2669
  • DOI: 10.1681/asn.2016121356
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Natriuretic peptide receptor guanylyl cyclase-A pathway counteracts glomerular injury evoked by aldosterone through p38 mitogen-activated protein kinase inhibition. (2017)
  • Author(s): Y. Kato, K. Mori, M. Kasahara, K. Osaki, A. Ishii, K.P. Mori, N. Toda, S. Ohno, T. Kuwabara, T. Tokudome, I. Kishimoto, M.A. Saleem, T. Matsusaka, K. Nakao, M. Mukoyama, M. Yanagita, H. Yokoi
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 46624-46624
  • DOI: 10.1038/srep46624
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] 腎臓病ガイドライン総まとめ：低用量カルペリチド (2017)
  • Author(s): 森 潔、山田博之
  • Journal Title: 月刊薬事 . 株式会社じほう (東京) Volume: 59 Pages: 40-43
• [Journal Article] Ablation of the N-type calcium channel ameliorates diabetic nephropathy with improved glycemic control and reduced blood pressure. (2016)
  • Author(s): Ohno S, Yokoi H, Mori K, Kasahara M, Kuwahara K, Fujikura J, Naito M, Kuwabara T, Imamaki H, Ishii A, Saleem MA, Numata T, Mori Y, Nakao K, Yanagita M, Mukoyama M
  • Journal Title: Sci Rep Volume: 6 Issue: 1 Pages: 27192-27192
  • DOI: 10.1038/srep27192
  • NAID: 120007100128
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Lipocalin 2 prevents intestinal inflammation by enhancing phagocytic bacterial clearance in macrophages (2016)
  • Author(s): Toyonaga T, Matsuura M, Mori K, Honzawa Y, Minami N, Yamada S, Kobayashi T, Hibi T, Nakase H
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 35104-35104
  • DOI: 10.1038/srep35014
  • NAID: 120006318859
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] NEW CONCEPT CONCERNING THE MECHANISM OF ALBUMINURIA IN THE EARLY STAGE OF DIABETIC NEPHROPATHY: INCREASED GLOMERULAR FILTRATION WITH PARTIAL COMPENSATION BY INCREASED TUBULAR REABSORPTION (2017)
  • Author(s): Keita P. Mori, Hideki Yokoi, Masato Kasahara, Takashige Kuwabara, Hirotaka Imamaki, Akira Ishii, Kazuwa Nakao, Motoko Yanagita, Masashi Mukoyama, and Kiyoshi Mori
  • Organizer: 54TH ERA-EDTA CONGRESS
  • Int'l Joint Research
• [Presentation] Crucial Role of Mesangial Cell-derived Connective Tissue Growth Factor in Anti-Glomerular Basement Membrane Glomerulonephritis (2017)
  • Author(s): Hideki Yokoi, Naohiro Toda, Kiyoshi Mori, Masato Kasahara, Akira Ishii, Keita P. Mori, Takashige Kuwabara, Taiji Matsusaka, Masashi Mukoyama, Motoko Yanagita
  • Organizer: 54TH ERA-EDTA CONGRESS
  • Int'l Joint Research
• [Presentation] 2017年尿中NGALの保険適応により、日本のAKI診療は変わるのか (2017)
  • Author(s): 森 潔
  • Organizer: 第60回日本腎臓学会学術総会
• [Remarks] 静岡県立大学教員データベース・教員情報詳細・森 潔
  • URL: http://db.u-shizuoka-ken.ac.jp/show/prof656.html