In the multi-step differentiation protocol used to generate pancreatic endocrine cells from human pluripotent stem cells, the induction of NGN3+ endocrine precursors from the PDX1+/NKX6.1+ pancreatic endoderm is crucial for efficient endocrine cell production. Here, we demonstrate that transient, not prolonged FOXO1 inhibition results in enhanced NGN3+ endocrine precursors and hormone-producing cell production. FOXO1 inhibition does not directly induce NGN3 expression but stimulates PDX1+/NKX6.1+ cell proliferation. NOTCH activity, whose suppression is important for Ngn3 expression, is not suppressed but Wnt signaling is stimulated by FOXO1 inhibition. Reversely, Wnt inhibition suppresses the effects of FOXO1 inhibitor. These findings indicate that FOXO1 and Wnt are involved in regulating the proliferation of PDX1+/NKX6.1+ pancreatic endoderm that gives rise to NGN3+ endocrine precursors.

在用于从人类多能干细胞生成胰腺内分泌细胞的多步分化方案中，从 PDX1+/NKX6.1+ 胰腺内胚层诱导 NGN3+ 内分泌前体对于有效的内分泌细胞生产至关重要。在这里，我们证明了短暂的，而不是长期的 FOXO1 抑制导致 NGN3+ 内分泌前体和产生激素的细胞产生增强。FOXO1 抑制不会直接诱导 NGN3 表达，但会刺激 PDX1+/NKX6.1+ 细胞增殖。NOTCH 活性，其抑制对 Ngn3 表达很重要，不会被抑制，但 Wnt 信号传导受到 FOXO1 抑制的刺激。相反，Wnt 抑制抑制 FOXO1 抑制剂的作用。这些发现表明 FOXO1 和 Wnt 参与调节 PDX1+/NKX6 的增殖。