The insulin and insulin-like growth factor-1 (IGF-1) receptors are important for the growth and development of embryonic tissues. To directly define their roles in the maintenance of pluripotency and differentiation of stem cells, we knocked out both receptors in induced pluripotent stem cells (iPSCs). iPSCs lacking both insulin and IGF-1 receptors (double knockout, DKO) exhibited preserved pluripotency potential despite decreased expression of transcription factors Lin28a and Tbx3 compared to control iPSCs. While embryoid body and teratoma assays revealed an intact ability of DKO iPSCs to form all three germ layers, the latter were composed of primitive neuroectodermal tumor-like cells in the DKO group. RNA-seq analyses of control vs DKO iPSCs revealed differential regulation of pluripotency, developmental, E2F1, and apoptosis pathways. Signaling analyses pointed to downregulation of the AKT/mTOR pathway and upregulation of the STAT3 pathway in DKO iPSCs in the basal state and following stimulation with insulin/IGF-1. Directed differentiation toward the three lineages was dysregulated in DKO iPSCs, with significant downregulation of key markers (Cebpα, Fas, Pparγ, and Fsp27) in adipocytes and transcription factors (Ngn3, Isl1, Pax6, and Neurod1) in pancreatic endocrine progenitors. Furthermore, differentiated pancreatic endocrine progenitor cells from DKO iPSCs showed increased apoptosis. We conclude that insulin and insulin-like growth factor-1 receptors are indispensable for normal lineage development and perturbations in the function and signaling of these receptors leads to upregulation of alternative compensatory pathways to maintain pluripotency.

胰岛素和胰岛素样生长因子-1 (IGF-1) 受体对胚胎组织的生长和发育很重要。为了直接确定它们在维持干细胞多能性和分化中的作用，我们敲除诱导多能干细胞 (iPSC) 中的两种受体。尽管转录因子Lin28a和Tbx3的表达降低，但缺乏胰岛素和 IGF-1 受体（双敲除，DKO）的 iPSC 仍表现出保留的多能性潜力与对照 iPSC 相比。虽然胚状体和畸胎瘤检测显示 DKO iPSCs 具有形成所有三个胚层的完整能力，但后者由 DKO 组中的原始神经外胚层肿瘤样细胞组成。对照与 DKO iPSC 的 RNA-seq 分析揭示了多能性、发育、E2F1 和细胞凋亡途径的差异调节。信号分析表明，在 DKO iPSCs 基础状态和胰岛素/IGF-1 刺激后，AKT/mTOR 通路下调和 STAT3 通路上调。在 DKO iPSC 中，向三个谱系的定向分化失调，关键标志物（Cebpα、Fas、Pparγ和Fsp27 ）显着下调）在胰腺内分泌祖细胞中的脂肪细胞和转录因子（Ngn3、Isl1、Pax6和Neurod1 ）中。此外，来自 DKO iPSC 的分化胰腺内分泌祖细胞显示出凋亡增加。我们得出结论，胰岛素和胰岛素样生长因子-1 受体对于正常谱系发育是必不可少的，并且这些受体的功能和信号传导的扰动导致替代补偿途径的上调以维持多能性。