Master regulatory factors for regeneration and EMT of kidney tubular cells

• Project/Area Number: 15K09273
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Kidney internal medicine
• Research Institution: Keio University
• Principal Investigator: Monkawa Toshiaki 慶應義塾大学, 医学部(信濃町), 教授 (80286484)
• Co-Investigator(Renkei-kenkyūsha): Ko Minoru 慶應義塾大学, 医学部, 教授 (50631199)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ES細胞 / iPS細胞 / 腎臓尿細管 / miRNA / 転写因子 / micro RNA / 腎線維化 / 尿細管 / 再生 / 腎臓 / 尿細管細胞 / EMT

Outline of Final Research Achievements

Based on in-silico analysis, we have identified the first set of four transcription factors to enhance the differentiation from hPSCs into SIX2+SALL1+ nephron progenitor cells (NPCs) with 92% efficiency within 2 days of reprogramming. The second set of four transcription factors converted SIX2+ NPCs into kidney organoids which contain multi-segmented nephron structures with characteristics of podocytes, proximal and distal tubules on day 14. This novel method using synthetic mRNAs might provide safe cellular sources without genome modifications for kidney regenerative therapies as wells as for disease modeling and drug screening in vitro.
Overexpression of miR-363 induced mesenchymal phenotypes with loss of epithelial phenotypes in human kidney tubular HKC-8 cells. We identified TWIST/canonical WNT pathway as the downstream effecter of miR-363, and inhibition of canonical WNT by small molecule, IWR-1, attenuated EMT induced by miR-363.

Research Products

• [Journal Article] miR-363 induces transdifferentiation of human kidney tubular cells to mesenchymal phenotype (2016)
  • Author(s): Morizane R, Fujii S, Monkawa T, Hiratsuka K, Yamaguchi S, Homma K, Itoh H.
  • Journal Title: Clin Exp Nephrol Volume: 20 Issue: 3 Pages: 394-401
  • DOI: 10.1007/s10157-015-1167-2
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Multi-segmented kidney organoids derived from human ES cells by stepwise transcription factor administration (2017)
  • Author(s): Ken Hiratsuka, Toshiaki Monkawa, Shintaro Yamaguchi, Ryuji Morizane, Shigeru B.H. Ko, Hiroshi Itoh, Minoru S.H. Ko
  • Organizer: Kidney Week 2017(ASN)
  • Int'l Joint Research
• [Presentation] A novel method to differentiate human ES cells into renal tubule-like cells by a combination of transcription factors administration (2016)
  • Author(s): Ken Hiratsuka, Toshiaki Monkawa, Shintaro Yamaguchi, Ryuji Morizane, Shigeru B.H. Ko, Hiroshi Itoh, Minoru S.H. Ko
  • Organizer: Kidney Week2016(ASN)
  • Place of Presentation: シカゴ（米国）
  • Year and Date: 2016-11-15
  • Int'l Joint Research
• [Presentation] ヒト尿細管マスター制御因子の検索と、転写因子をコードする修飾合成mRNAを用いたヒト多能性幹細胞から尿細管様細胞への分化誘導方法の開発 (2016)
  • Author(s): 平塚 健, 門川俊明, 秋山智彦, 中武悠樹, Sravan Kumar Goparaju, 小林紗恵子, 納富奈々, 木村寛美, 山口慎太郎, 森實隆司, 鈴木さゆり, 洪 繁, 洪 実, 伊藤 裕
  • Organizer: 第7回分子腎臓フォーラム
  • Place of Presentation: ベルサール八重洲（東京都中央区）
• [Presentation] The Direct Differentiation Method of Renal Tubular Cells by Synthetic mRNAs of Transcription Factors Identi ed from TF-Inducible Human ES Bank (2015)
  • Author(s): Ken Hiratsuka, Toshiaki Monkawa, Shintaro Yamaguchi, Ryuji Morizane, Shigeru B.h. Ko, Minoru S.h. Ko, Hiroshi Itoh
  • Organizer: Kidney Week 2015
  • Place of Presentation: San Diego, CA, USA
  • Year and Date: 2015-11-06
  • Int'l Joint Research