Pre-adaptation of Stem Cell-Derived Islet Organoids to Hypoxia via Zinc Transportation Inhibition Drives Rapid Neo-vascularization

Human stem cell-derived islet organoids (SC-islets) hold great potential for diabetes treatment, but their clinical application is hindered by immaturity and ischemia-induced dysfunction post transplantation. Hypoxia-driven angiogenesis via the HIF1A-VEGFA axis is a common adaptation, but the metabolic fragility of SC-islet ß cell leads to early functional damage and suppressed VEGFA release, thereby delaying vascularization and causing graft loss. The key challenge in SC-islet transplantation is how to prevent hypoxia-induced stress and promote rapid neo-vascularization. To reconcile this discrepancy, we targeted zinc transportation using a chemical inhibitor to enhance SC-islet maturation, hypoxia resistance and vascularization. We found that excessive zinc in SC-islet ß-cells induces oxidative modification that inhibits AMPK activity. Chemical inhibition of zinc transportation activates AMPK, simultaneously enhances functional maturation, improves hypoxia resistance and increases VEGFA expression to facilitate endothelial cell integration. In diabetic animal models, this approach significantly improved hypoxia resistance, accelerated neo-vascularization, and enhanced glycemic control. Our findings demonstrate that chemical inhibition of zinc transportation boosts SC-islet functional competence, offering a potential strategy to advance the paradigm of functional pre-adaptation to stress in regenerative medicine. Overall design: We generated hESCs reporter cell line (MEL1 Nkx6.1:linker2a:mCherry; INSGFP/w) that enable to precisely and dynamically trace SC-islets during differentiated stages. SC-islets by sorting out GFP and mCherry double-positive SC-ß cells with high INS and NKX6.1 expression