严重的肠道副作用针对人类癌症分化治疗的NOTCH-HES1通路，使得了解该通路在人体器官水平上是必要的。因此，我们将HES1-/-突变体内源性引入到人类胚胎干细胞（hESCs）中，并将其分化成人类肠道器官样体（HIO）。当它们分化成定义性内胚层和后肠时，HES1-/- hESCs保留了ES细胞的特性，并且表现出与野生型hESCs相似的基因表达模式。在形成HES1-/-灯腔的过程中，我们注意到间充质细胞的发育受到了抑制，此外，分泌上皮的分化也增加了。RNA-Seq揭示了间质细胞发育受到抑制可能是由于WNT5A信号通路的下调导致的。 在肠纤维细胞系CCD-18Co中过表达HES1和沉默WNT5A表明，HES1参与激活WNT5A诱导的成纤维细胞生长和迁移，暗示了在上皮间质相互作用中可能涉及到Notch通路。我们的结果促进了更精确的基础分子机制的识别，展示了HES1信号通路在人类肠黏膜的基质和上皮发育中具有不同的功能。 © 2023 Elsevier Inc.发表。

Serious intestinal side-effects that target the NOTCH-HES1 pathway in human cancer differentiation therapy make it necessary to understand the pathway at the human organ level. Herein, we endogenously introduced HES1-/- mutations into human embryonic stem cells (hESCs) and differentiated them into human intestinal organoids (HIO). The HES1-/- hESCs retained ES cell properties and showed gene expression patterns similar to those of wild-type hESCs when they differentiated into definitive endoderm and hindgut. During the formation of the HES1-/- lumen we noted an impaired development of mesenchymal cells in addition to the increased differentiation of secretory epithelium. RNA-Seq revealed that inhibited development of the mesenchymal cells may have been due to a downregulation of WNT5A signaling. Overexpression of HES1 and silencing of WNT5A in the intestinal fibroblast cell line CCD-18Co indicated that HES1 was involved in the activation of WNT5A-induced fibroblast growth and migration, suggesting the likelihood of the Notch pathway in epithelial-mesenchymal crosstalk. Our results facilitated the identification of more precise underlying molecular mechanisms displaying distinct roles in HES1 signaling in stromal and epithelial development in human intestinal mucosa.Copyright © 2023. Published by Elsevier Inc.