Stomach-derived human insulin-secreting organoids ameliorate diabetes

Adult stem cells represent an invaluable resource that can be harnessed for therapeutic tissue repair. Gut stem cells are accessible by biopsy and grow indefinitely in culture as organoids or cell lines. Human fetal gut can generate rare insulin-secreting cells. However, the short lifespan of gut cells, amounting to only days in situ, calls into question the feasibility of producing stable and durable gut insulin-secreting cells as a potential engraftable therapeutic. Here, we show that cultured human stomach stem cells can be directed to generate pancreatic islet-like organoids containing long-lived gastric insulin-secreting (GINS) cells that resemble pancreatic b-cells in molecular hallmarks and function. After sequential activation of the inducing factors NGN3 and PDX1-MAFA, gastric stem cells passed through a SOX4High endocrine and a GalaninHigh precursor state before adopting the b-cell fate, at efficiencies exceeding 80%. GINS cells acquired glucose-stimulated insulin secretion 10 days post differentiation and restored glucose homeostasis for over 100 days in diabetic mice after transplantation. This study establishes a promising approach to procuring autologous human insulin producers for diabetes treatment, and further expands the already considerable therapeutic opportunities for gut stem cells. Overall design: Single-cell RNA-seq on (1) human gastric stem cells derived from corpus stomach, (2) endocrine progenitors, (3) GINS precursors, (4) corpus GINS organoids, (5) antral GINS organoids, and (6) corpus GINS cells after transplantation. Please note that the mixed sample represents: Human gastric stem cells (hGSCs) derived from corpus stomach Endocrine progenitors differentiated from corpus hGSCs GINS precursors differentiated from corpus hGSCs GINS organoids differentiated from corpus hGSCs To demultiplex, the CMO IDs for the 10x multiplex are: Sample1 = CMO01 Sample2 = CMO02 Sample3 = CMO03 Sample4 = CMO04