Introduction & Objective: Pancreatic beta cell neogenesis is a phenomenon whereby new beta cells are generated from other pancreatic progenitor cells. Although human beta-cell neogenesis has been directly observed, questions remain as to exactly how beta-cell neogenesis is controlled and whether it can be induced with unknown genes.
Methods: A genome-wide CRISPR screen was employed on the REPB-PANC-1 cells with a human lentiviral genome-wide CRISPR knockout library (GeCKO v2). (Reporter construct: Rat insulin promoter 3.1 (RIP)-EGFP-P2A-Blasticidin-S deaminase (BSD). Statistical analyses were performed by unpaired or paired tests as indicated using the Prism 8 software.
Results: We demonstrated that the loss-of-function of a single gene,ALDH3B2, in pancreatic duct cells is sufficient to induce bonafide cell trans-differentiation from human pancreatic duct cells to beta-like cells. The gene expression of ofPDX1, INS, and SLC2A1 were significantly upregulated in the mutant primary pancreatic ductal cells; We demonstrated that the trans-differentiated human pancreatic beta-like cells are responsive to glucose challengein vivo and able to significantly lower blood glucose in diabetic animal models.
Conclusion: Overall, our whole study led to the discovery ofALDH3B2 as a beta-cell neogenesis regulator. Our finding was conducted using human pancreatic duct cell lines or primary human pancreatic duct cells, which provides a good basis for translating into future human diabetes therapeutics.
J. Li: None. P. Yi: None.
