Renal tubular cells derived from human ES cells and hepatocytes by transcription factor administration

• Project/Area Number: 16H07177
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Kidney internal medicine
• Research Institution: Keio University
• Principal Investigator: Hiratsuka Ken 慶應義塾大学, 医学部(信濃町), 助教 (80594481)
• Research Collaborator: KO Minoru 慶應義塾大学, 医学部, 教授 (50631199)
• Project Period (FY): 2016-08-26 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: ヒトES細胞 / 転写調節因子 / リプログラミング / 腎臓再生 / 多能制幹細胞 / 転写因子 / 再生医療 / 腎尿細管細胞 / ヒト多能性幹細胞 / 肝細胞 / 尿細管細胞

Outline of Final Research Achievements

We utilized the transcription factor (TF)-inducible human ES lines, analyzed correlation of gene expression response to the induction of human TFs with tissue-specific gene expression in silico, and identified candidate TFs for differentiating towards renal lineages. Based on in-silico analysis, we have identified a set of four TFs that induce nephron progenitor cells (NPCs) and another set of four TFs that help differentiate toward pretubular aggregate-like cells. Two days after the transfection of the first set of four TFs together into hESCs, NPCs were induced efficiently (~92%). Subsequent administration of the second set of four TFs transfection induced differentiation into pretubular aggregate-like cells. On day 14, epithelial characteristic changes and multi-segmented kidney structures containing podocyte, proximal tubules, and distal tubules were observed in 3D cultures. RNA-Seq analysis also showed the global expression profile closely resembled human kidney profile.

Research Products

• [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(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: Chicago, IL
  • 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
  • Year and Date: 2016-09-03