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Single nuclei multiomics of human stem cell-derived islets identifies deficiencies in lineage specification

ABSTRACT: We profiled the transcritpome and ATAC profiles of human pancreatic islets generated from pluripotent stem cells. Multiomic profiling was also performed on primary human islets and in vivo matured SC-islets for comparision. We catalogued the ATAC associated signatures for each cell types in SC-islets and compared them to their human primiary islet counterparts. In vivo maturation of SC-islets were also compared with in vitro SC-islets. In this study, we identified key regulators associated with islet identity during differentiation and maturation. Gene manipulation of CTCF affects differentiating SC-islet cell fate to enteroendocrine-like lineage. ARID1B knockdown caueses islet cells to present mature signatures. These gene altered SC-islets were also sequenced.

ORGANISM(S): Homo sapiens

PROVIDER: GSE199636 | GEO | 2023/03/25

REPOSITORIES: GEO

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Vascularization of pluripotent stem cell-derived islet organoids improves β-cell function

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Restoration of PKM1 improves functional maturation of stem-cell derived-b by regulating PEP metabolism

Project description:In stem cell-derived β (SC-β) cells, metabolic abnormalities persist as obstacles to glucose responsiveness and functional maturation, with the primary metabolic bottlenecks yet to be identified. This study demonstrated that restoring pyruvate kinase 1 (PKM1) re-established pyruvate kinase activity, effectively reversing the SC-β-cell identity loss and functional impairment associated with phosphoenolpyruvate (PEP) accumulation. 13C-glucose labeling metabolomics revealed a disruption in glucose-stimulated metabolite production beginning at the glycolytic PEP stage in stem cell-derived islets (SC-islets). Uniquely, elevated PEP prevented glycolytic metabolite-dependent increase of exocytosis and significantly increased intracellular calcium levels through a KATP channel-independent pathway, even under basal glucose condition. Furthermore, exposure to PEP led to downregulated expression of genes involved in the TCA cycle and respiratory electron transport. This PEP accumulation in SC-islets was associated with markedly reduced pyruvate kinase activity. Overexpression of PKM1 rescued the transcriptional changes induced by PEP accumulation and improved both glucose-stimulated calcium responses and insulin secretion. These findings suggest that PKM1 plays a pivotal role in promoting SC-β cell differentiation and functional maturation by regulating PEP metabolism, offering potential improvements for SC-β cell replacement in diabetes treatment.

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Project description:During pancreas development, endocrine progenitors differentiate into the islet-cell subtypes, which undergo further functional maturation in postnatal islet development. In islet b-cells, genes involved in glucose-stimulated insulin secretion are activated and glucose exposure increases the insulin response as b-cells mature. Here, we investigated the role of H3K4 trimethylation in endocrine cell differentiation and functional maturation by disrupting TrxG complex histone methyltransferase activity in mouse endocrine progenitors. In the embryo, genetic inactivation of TrxG component Dpy30 in NEUROG3+ cells did not affect the number of endocrine progenitors or endocrine cell differentiation. H3K4 trimethylation was progressively lost in postnatal islets and the mice displayed elevated non-fasting and fasting glycemia, as well as impaired glucose tolerance by postnatal day 24. Although postnatal endocrine cell proportions were equivalent to controls, islet RNA-sequencing revealed a downregulation of genes involved in glucose-stimulated insulin secretion and an upregulation of immature b-cell genes. Comparison of histone modification enrichment profiles in NEUROG3+ endocrine progenitors and mature islets suggested that genes downregulated by loss of H3K4 trimethylation more frequently acquire active histone modifications during maturation. Taken together, these findings suggest that H3K4 trimethylation is required for the activation of genes involved in the functional maturation of pancreatic islet endocrine cells.

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