2. Academic Validation
  3. CDK8/19 inhibition plays an important role in pancreatic β-cell induction from human iPSCs

CDK8/19 inhibition plays an important role in pancreatic β-cell induction from human iPSCs

  • Stem Cell Res Ther. 2023 Jan 5;14(1):1. doi: 10.1186/s13287-022-03220-4.
Kensuke Sakuma 1 2,  Noriko Tsubooka-Yamazoe 3 4,  Kiyohiro Hashimoto 5,  Nozomu Sakai 6,  Shinya Asano 7,  Saori Watanabe-Matsumoto 4 8,  Takeshi Watanabe 5,  Bunnai Saito 6,  Hirokazu Matsumoto 4 9,  Hikaru Ueno 3 4,  Ryo Ito 3 4,  Taro Toyoda 10 11
Affiliations

Affiliations

  • 1 iPSC-Derived Pancreatic Islet Cell (iPIC) Therapy Department, Orizuru Therapeutics Inc., Fujisawa, Kanagawa, 251-8555, Japan. [email protected].
  • 2 Takeda-CiRA Joint Program for iPS Cell Applications (T-CiRA), Fujisawa, Kanagawa, 251-8555, Japan. [email protected].
  • 3 iPSC-Derived Pancreatic Islet Cell (iPIC) Therapy Department, Orizuru Therapeutics Inc., Fujisawa, Kanagawa, 251-8555, Japan.
  • 4 Takeda-CiRA Joint Program for iPS Cell Applications (T-CiRA), Fujisawa, Kanagawa, 251-8555, Japan.
  • 5 Drug Safety Research and Evaluation Group, Takeda Pharmaceutical Company Limited, Kanagawa, 251-8555, Japan.
  • 6 Drug Discovery Sciences, Takeda Pharmaceutical Company Limited, Kanagawa, 251-8555, Japan.
  • 7 Integrated & Translational Science, Axcelead Drug Discovery Partners, Inc., Fujisawa, Kanagawa, 251-8555, Japan.
  • 8 Department of Life Science Frontiers, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, 606-8507, Japan.
  • 9 T-CiRA Discovery and Innovation, Takeda Pharmaceutical Company Limited, Kanagawa, 251-8555, Japan.
  • 10 Takeda-CiRA Joint Program for iPS Cell Applications (T-CiRA), Fujisawa, Kanagawa, 251-8555, Japan. [email protected].
  • 11 Department of Life Science Frontiers, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, 606-8507, Japan. [email protected].
PMID: 36600289 DOI: 10.1186/s13287-022-03220-4
Abstract

Background: Transplantation of differentiated cells from human-induced pluripotent stem cells (hiPSCs) holds great promise for clinical treatments. Eliminating the risk factor of malignant cell transformation is essential for ensuring the safety of such cells. This study was aimed at assessing and mitigating mutagenicity that may arise during the Cell Culture process in the protocol of pancreatic islet cell (iPIC) differentiation from hiPSCs.

Methods: We evaluated the mutagenicity of differentiation factors used for hiPSC-derived pancreatic islet-like cells (iPICs). We employed Ames mutagenicity assay, flow cytometry analysis, immunostaining, time-resolved fluorescence resonance energy transfer-based (TR-FRET) cell-free dose-response assays, single-cell RNA-sequencing and in vivo efficacy study.

Results: We observed a mutagenic effect of activin receptor-like kinase 5 inhibitor II (ALK5iII). ALK5iII is a widely used β-cell inducer but no other tested ALK5 inhibitors induced β-cells. We obtained kinase inhibition profiles and found that only ALK5iII inhibited cyclin-dependent kinases 8 and 19 (CDK8/19) among all ALK5 inhibitors tested. Consistently, CDK8/19 inhibitors efficiently induced β-cells in the absence of ALK5iII. A combination treatment with non-mutagenic ALK5 Inhibitor SB431542 and CDK8/19 inhibitor senexin B afforded generation of iPICs with in vitro cellular composition and in vivo efficacy comparable to those observed with ALK5iII.

Conclusion: Our findings suggest a new risk mitigation approach for cell therapy and advance our understanding of the β-cell differentiation mechanism.

Keywords

Activin receptor-like kinase 5 inhibitor II; CDK8/19 inhibitors; Human-induced pluripotent stem cells; Mutagenicity; Pancreatic islet cell.

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