The differentiation of pluripotent stem cells has been used to study disease mechanisms and development. We previously described a method for differentiating human pluripotent stem cells (hPSCs) into salivary gland epithelial progenitors (SGEPs). Here, cystic fibrosis transmembrane conductance regulator (CFTR) knockout hPSCs were differentiated into SGEPs derived from CFTR knockout hESCs (CF-SGEPs) using the same protocol to investigate whether the hPSC-derived SGEPs can model the characteristics of CF. CF-a disease that affects salivary gland (SG) function-is caused by mutations of the CFTR gene. Firstly, we successfully generated CFTR knockout hPSCs with reduced CFTR protein expression using the CRISPR-Cas9 system. After 16 days of differentiation, the protein expression of CFTR decreased in SGEPs derived from CFTR knockout hESCs (CF-SGEPs). RNA-Seq revealed that multiple genes modulating SG development and function were down-regulated, and positive regulators of inflammation were up-regulated in CF-SGEPs, correlating with the salivary phenotype of CF patients. These results demonstrated that CFTR suppression disrupted the differentiation of hPSC-derived SGEPs, which modeled the SG development of CF patients. In summary, this study not only proved that the hPSC-derived SGEPs could serve as manipulable and readily accessible cell models for the study of SG developmental diseases but also opened up new avenues for the study of the CF mechanism.

中文翻译：

---

囊性纤维化跨膜电导调节器敲除人类多能干细胞向唾液腺上皮祖细胞的分化和表征。

多能干细胞的分化已用于研究疾病机制和发展。我们之前描述了一种将人多能干细胞 (hPSC) 分化为唾液腺上皮祖细胞 (SGEP) 的方法。在这里，使用相同的方案将囊性纤维化跨膜电导调节器 (CFTR) 敲除 hPSC 分化为源自 CFTR 敲除 hESC 的 SGEP (CF-SGEP)，以研究 hPSC 衍生的 SGEP 是否可以模拟 CF 的特征。CF 是一种影响唾液腺 (SG) 功能的疾病，由CFTR基因突变引起。首先，我们使用 CRISPR-Cas9 系统成功生成了 CFTR 敲除 hPSC，其 CFTR 蛋白表达减少。分化 16 天后，CFTR 敲除 hESC 衍生的 SGEP（CF-SGEP）中 CFTR 的蛋白表达下降。RNA-Seq 显示，CF-SGEP 中调节 SG 发育和功能的多个基因下调，炎症正向调节因子上调，这与 CF 患者的唾液表型相关。这些结果表明，CFTR 抑制破坏了 hPSC 衍生的 SGEP 的分化，从而模拟了 CF 患者的 SG 发育。总之，本研究不仅证明hPSC衍生的SGEP可以作为SG发育疾病研究的可操作且易于获得的细胞模型，而且为CF机制的研究开辟了新途径。