Project-2: Modeling TE birth defects in animals

项目2|项目总结 未能将胎儿前肠分离为不同的气管和食道(TE)可能导致一系列生命- 有威胁的气管-食道缺陷(TEDS)。TEDS的遗传病因知之甚少，而且有风险 只有12%的TED患者知道有变异。即使在已知致病突变的情况下 与发育转录因子一样，它们如何导致TED尚不清楚，因为直到最近， TE的形态发生机制尚不清楚。我们在上一次获奖中取得了重大进展。vbl.使用 结合非洲爪哇和小鼠胚胎学，我们阐明了驱动TE的保守细胞事件 形态发生的研究表明，干扰任何一步都可以产生类似TED的表型。我们用的是非洲爪哇 CRISPR筛选以验证患者基因组序列中可能导致TED的变异。一大专业 研究发现，Hedgehog(HH)信号的下游和发育中的TF是由内吞体介导的 膜重塑是将前肠分离成不同的TE管所必需的。与此一致的是，基因组 对项目1中185名TED患者的测序显示，在 膜/囊泡运输基因，包括内吞适配器ITSN1，在初步数据中 已证明是非洲爪哇形态发生所必需的。Clear财团项目2的目标是 在动物模型中用细胞分辨率确定TED的发育基础。我们将：确定 内小体运输调节TE形态发生的机制(Aim1)。检验以下假设 发育中的转录因子控制效应蛋白的细胞特异性表达，如内小体转运 机械或货物(AIM2，与项目3合作)。评估患者变种在动物身上的致病性 检验内胚层病变可能是TEDS的主要原因这一挑衅性假设(Aim3)。

PROJECT 2 | PROJECT SUMMARY Failure to separate the fetal foregut into distinct trachea and esophagus (TE) can result in a spectrum of life- threatening tracheoesophageal defects (TEDs). The genetic etiology of TEDs is poorly understood, and risk variants are known in only 12% of TED patients. Even in cases where the causative mutations are known such as in developmental transcription factors (TFs), how they result in TEDs is unclear because up until recently the mechanisms of TE morphogenesis were ill defined. We made significant progress in the previous award. Using a combination of Xenopus and mouse embryology, we elucidated the conserved cellular events driving TE morphogenesis and showed that disrupting any step can result in TED-like phenotypes. We used Xenopus CRISPR screens to validate potential TED-causing variants from patient genome sequences. One major discovery was that downstream of Hedgehog (HH) signaling and developmental TFs, endosome-mediated membrane remodeling is required to separate the foregut into distinct TE tubes. Consistent with this, genome sequencing of 185 TED patients from Project 1 revealed an enrichment of damaging variants in membrane/vesicular trafficking genes including the endocytic adaptor ITSN1, which in preliminary data we demonstrated is required for Xenopus TE morphogenesis. The goal of the CLEAR consortium Project 2 is to define the developmental basis of TED with cellular resolution in animal models. We will: Determine the mechanism by which endosome trafficking regulates TE morphogenesis (Aim1). Test the hypothesis that developmental TFs control the cell-specific expression of effector proteins such as endosome trafficking machinery or cargo (Aim2, in collaboration with Project 3). Assess the pathogenicity of patient variants in animals testing the provocative hypothesis that endosomeopathies might be a major cause of TEDs (Aim3).