AIM The loss of insulin-secreting β-cells, ultimately characterizing most diabetes forms, demands the development of cell replacement therapies. The common endpoint for all ex vivo strategies is transplantation into diabetic patients. However, the effects of hyperglycaemia environment on the transplanted cells were not yet properly assessed. Thus, the main goal of this study was to characterize global effect of brief and prolonged in vivo hyperglycaemia exposure on the cell fate acquisition and maintenance of transplanted human pancreatic progenitors. METHODS To rigorously study the effect of hyperglycaemia, in vitro differentiated human-induced pluripotent stem cells (hiPSC)-derived pancreatic progenitors were xenotransplanted in normoglycaemic and diabetic NSG rat insulin promoter (RIP)-diphtheria toxin receptor (DTR) mice. The transplants were retrieved after 1-week or 1-month exposure to overt hyperglycaemia and analysed by large-scale microscopy or global proteomics. For this study we pioneer the use of the NSG RIP-DTR system in the transplantation of hiPSC, making use of its highly reproducible specific and absolute β-cell ablation property in the absence of inflammation or other organ toxicity. RESULTS Here we show for the first time that besides the presence of an induced oxidative stress signature, the cell fate and proteome landscape response to hyperglycaemia was different, involving largely different mechanisms, according to the period spent in the hyperglycaemic environment. Surprisingly, brief hyperglycaemia exposure increased the bihormonal cell number by impeding the activity of specific islet lineage determinants. Moreover, it activated antioxidant and inflammation protection mechanisms signatures in the transplanted cells. In contrast, the prolonged exposure was characterized by decreased numbers of hormone + cells, low/absent detoxification signature, augmented production of oxygen reactive species and increased apoptosis. CONCLUSION Hyperglycaemia exposure induced distinct, period-dependent, negative effects on xenotransplanted human pancreatic progenitor, affecting their energy homeostasis, cell fate acquisition and survival.

中文翻译：

---

体内高血糖暴露引起人体胰腺祖细胞分化的明显的周期依赖性效应，这是通过氧化应激传递的。

目的胰岛素分泌性β细胞的丢失最终是大多数糖尿病形式的特征，需要开发细胞替代疗法。所有体外策略的共同终点是移植到糖尿病患者中。但是，高血糖环境对移植细胞的影响尚未得到适当评估。因此，本研究的主要目的是表征短暂和长期体内高血糖暴露对移植的人类胰腺祖细胞的细胞命运获得和维持的总体影响。方法为了严格研究高血糖的作用，将体外分化的人诱导多能干细胞（hiPSC）衍生的胰腺祖细胞异种移植到正常血糖和糖尿病NSG大鼠胰岛素启动子（RIP）-白喉毒素受体（DTR）小鼠中。暴露于明显的高血糖症1周或1个月后取回移植物，并通过大规模显微镜或整体蛋白质组学进行分析。在这项研究中，我们率先在NSPS RIP-DTR系统的hiPSC移植中使用它，在没有炎症或其他器官毒性的情况下，利用其高度可重复的特异性和绝对β细胞消融特性。结果在这里，我们首次表明，除了在高血糖环境中所花费的时间以外，对高血糖的细胞命运和蛋白质组景观反应也不同，涉及的机制也大不相同。出乎意料的是，短暂的高血糖暴露通过阻碍特定胰岛谱系决定簇的活性而增加了激素细胞的数量。而且，它激活了移植细胞中的抗氧化剂和炎症保护机制。相反，长时间暴露的特征是激素+细胞数量减少，排毒特征低/不存在，氧反应性物质产量增加和细胞凋亡增加。结论高血糖暴露对异种移植的人类胰腺祖细胞具有明显的，周期依赖性的负面影响，影响其能量稳态，细胞命运获取和存活。