Human Enteroids, Colonoids, and iPSC derived HIO's to study CFTR-relatedDisorders

项目摘要 在这个提议中要检验的统一假设是突变CFTR的纠正和增强作用 鸟苷酸环化酶2C（GC-C）激动剂可以恢复肠道的液体稳态， 源自突变CFTR功能缺陷的慢性便秘。我们将用病人来检验这个假设。 来源于隐窝（肠样组织）的肠干细胞，以研究体外液体分泌; 多能干细胞（iPSC）衍生的人肠类器官（HIO）在小鼠肾被膜中， 用作研究体内液体分泌的模型。这些研究非常重要，因为（i） 通过利用人类模型解决了几种致命的人类疾病的病理学;（ii）它已经 临床相关性和影响;（iii）它是一个多学科项目，涵盖基础生物医学研究， 开发和药物发现使用个性化的人类干细胞培养。

PROJECT SUMMARY The unifying hypothesis to be tested in this proposal is that the mutant CFTR correcting and potentiating roles of guanylate cyclase 2C (GC-C)-agonists can restore fluid homeostasis of the gut in a subset of population with chronic constipation emanating from mutant CFTR functional deficit. We will test this hypothesis using patient derived intestinal stem cells derived from crypts (enteroids) to study fluid secretion in vitro; and the induced pluripotent stem cells (iPSCs) derived human intestinal organoids (HIOs) in mouse kidney capsule and be used as models to study fluid secretion in vivo. The proposed studies are highly significant because (i) it addresses the pathologies of several deadly human diseases by utilizing models from humans; (ii) it has clinical relevance and implications; (iii) it is a multidisciplinary project covers basic biomedical studies, assay developments, and drug discovery using personalized human stem cell cultures.