p53 Inhibition in Pancreatic Progenitors Enhances the Differentiation of Human Pluripotent Stem Cells into Pancreatic β-Cells

• Published in: Stem cell reviews and reports Vol. 19; no. 4; pp. 942 - 952
• Main Authors: Aigha, Idil I., Abdelalim, Essam M.
• Format: Journal Article
• Language: English
• Published: New York Springer US 01.05.2023; Springer Nature B.V
• Subjects: Beta cells; Biomedical and Life Sciences; Biomedical Engineering and Bioengineering; Cell Biology; Cell differentiation; Cell Differentiation - genetics; Embryo cells; Glucagon; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; Humans; Life Sciences; Nkx6.1 protein; p53 Protein; Pancreas; Pluripotency; Pluripotent Stem Cells; Progenitor cells; Regenerative Medicine/Tissue Engineering; Stem Cells; Transcription factors; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; Islets; Monohormonal β-cells; hESCs; NKX6.1; Insulin; Differentiation protocol
• ISSN: 2629-3269; 2629-3277
• DOI: 10.1007/s12015-023-10509-1

The multipotent pancreatic progenitor cells (MPCs) co-expressing the transcription factors, PDX1 and NKX6.1, are the source of functional pancreatic β-cells. The aim of this study was to examine the effect of p53 inhibition in MPCs on the generation of PDX1 + /NKX6.1 + MPCs and pancreatic β-cell generation. Human embryonic stem cells (hESCs) were differentiated into MPCs and β-cells. hESC-MPCs (stage 4) were treated with different concentrations of p53 inhibitors, and their effect was evaluated using different approaches. NKX6.1 was overexpressed during MPCs specification. Inhibition of p53 using pifithrin-μ (PFT-μ) at the MPC stage resulted in a significant increase in the number of PDX1 + /NKX6.1 + cells and a reduction in the number of CHGA + /NKX6.1 − cells. Further differentiation of MPCs treated with PFT-μ into pancreatic β-cells showed that PFT-μ treatment did not significantly change the number of C-Peptide+ cells; however, the number of C-PEP+ cells co-expressing glucagon (polyhormonal) was significantly reduced in the PFT-μ treated cells. Interestingly, overexpression of NKX6.1 in hESC-MPCs enhanced the expression of key MPC genes and dramatically suppressed p53 expression. Our findings demonstrated that the p53 inhibition during stage 4 of differentiation enhanced MPC generation, prevented premature endocrine induction and favored the differentiation into monohormonal β-cells. These findings suggest that adding a p53 inhibitor to the differentiation media can significantly enhance the generation of monohormonal β-cells. Graphical Abstract