Remodeling ceramide homeostasis promotes functional maturation of human pluripotent stem cell-derived (3 cells

Human pluripotent stem cell -derived (3 cells (hPSC-(3 cells) show the potential to restore euglycemia. However, the immature functionality of hPSC-(3 cells has limited their efficacy in application. Here, by deciphering the continuous maturation process of hPSC-(3 cells post transplantation via single -cell RNA sequencing (scRNA-seq) and single -cell assay for transposase-accessible chromatin sequencing (scATAC-seq), we show that functional maturation of hPSC-(3 cells is an orderly multistep process during which cells sequentially undergo metabolic adaption, removal of negative regulators of cell function, and establishment of a more specialized transcriptome and epigenome. Importantly, remodeling lipid metabolism, especially downregulating the metabolic activity of ceramides, the central hub of sphingolipid metabolism, is critical for (3 cell maturation. Limiting intracellular accumulation of ceramides in hPSC-(3 cells remarkably enhanced their function, as indicated by improvements in insulin processing and glucose -stimulated insulin secretion. In summary, our findings provide insights into the maturation of human pancreatic (3 cells and highlight the importance of ceramide homeostasis in function acquisition.