2. Academic Validation
  3. Nicotinamide promotes pancreatic differentiation through the dual inhibition of CK1 and ROCK kinases in human embryonic stem cells

Nicotinamide promotes pancreatic differentiation through the dual inhibition of CK1 and ROCK kinases in human embryonic stem cells

  • Stem Cell Res Ther. 2021 Jun 25;12(1):362. doi: 10.1186/s13287-021-02426-2.
Yumeng Zhang 1 2,  Jiaqi Xu 1 2,  Zhili Ren 1 2,  Ya Meng 1 2 3,  Weiwei Liu 1 4,  Ligong Lu 3,  Zhou Zhou 5,  Guokai Chen 6 7 8
Affiliations

Affiliations

  • 1 Centre of Reproduction, Development and Aging, Faculty of Health Sciences, University of Macau, Macau SAR, China.
  • 2 Institute of Translational Medicine, Faculty of Health Sciences, University of Macau, Macau SAR, China.
  • 3 Zhuhai Precision Medical Center, Zhuhai People's Hospital, Jinan University, Zhuhai, Guangdong, China.
  • 4 Bioimaging and Stem Cell Core Facility, Faculty of Health Sciences, University of Macau, Macau SAR, China.
  • 5 State Key Laboratory of Cardiovascular Disease, Beijing Key Laboratory for Molecular Diagnostics of Cardiovascular Diseases, Diagnostic Laboratory Service, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
  • 6 Centre of Reproduction, Development and Aging, Faculty of Health Sciences, University of Macau, Macau SAR, China. [email protected].
  • 7 Institute of Translational Medicine, Faculty of Health Sciences, University of Macau, Macau SAR, China. [email protected].
  • 8 MoE Frontiers Science Center for Precision Oncology, University of Macau, Macau SAR, China. [email protected].
PMID: 34172095 DOI: 10.1186/s13287-021-02426-2
Abstract

Background: Vitamin B3 (nicotinamide) plays important roles in metabolism as well as in SIRT and PARP pathways. It is also recently reported as a novel kinase inhibitor with multiple targets. Nicotinamide promotes pancreatic cell differentiation from human embryonic stem cells (hESCs). However, its molecular mechanism is still unclear. In order to understand the molecular mechanism involved in pancreatic cell fate determination, we analyzed the downstream pathways of nicotinamide in the derivation of NKX6.1+ pancreatic progenitors from hESCs.

Methods: We applied downstream modulators of nicotinamide during the induction from posterior foregut to pancreatic progenitors, including niacin, PARP Inhibitor, SIRT inhibitor, CK1 Inhibitor and ROCK Inhibitor. The impact of those treatments was evaluated by quantitative Real-Time PCR, flow cytometry and immunostaining of pancreatic markers. Furthermore, CK1 isoforms were knocked down to validate CK1 function in the induction of pancreatic progenitors. Finally, RNA-seq was used to demonstrate pancreatic induction on the transcriptomic level.

Results: First, we demonstrated that nicotinamide promoted pancreatic progenitor differentiation in chemically defined conditions, but it did not act through either niacin-associated metabolism or the inhibition of PARP and SIRT pathways. In contrast, nicotinamide modulated differentiation through CK1 and ROCK inhibition. We demonstrated that CK1 inhibitors promoted the generation of PDX1/NKX6.1 double-positive pancreatic progenitor cells. shRNA knockdown revealed that the inhibition of CK1α and CK1ε promoted pancreatic progenitor differentiation. We then showed that nicotinamide also improved pancreatic progenitor differentiation through ROCK inhibition. Finally, RNA-seq data showed that CK1 and ROCK inhibition led to pancreatic gene expression, similar to nicotinamide treatment.

Conclusions: In this report, we revealed that nicotinamide promotes generation of pancreatic progenitors from hESCs through CK1 and ROCK inhibition. Furthermore, we discovered the novel role of CK1 in pancreatic cell fate determination.

Keywords

Casein kinase 1 (CK1); Human embryonic stem cells; Kinase inhibitor; Nicotinamide; Pancreatic progenitors; Rho-associated protein kinase (ROCK).

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