Transcriptional and epigenetic basis of lung epithelial cell fate

项目总结 肺上皮由不同类型的细胞组成，这是胚胎祖细胞经历 一系列精确的细胞命运决定。在受伤后，成功的修复需要恢复适当的细胞 命运。体内细胞命运生物学的概述和调控为基于细胞的生物学提供了希望 再生疗法。尽管有如此重要的基础和我们对细胞不断积累的知识 命运调节，肺细胞命运的表观遗传学基础-在本提案中定义为染色质状态和 它与转录因子结合的关系在很大程度上是未知的，但对于机制来说是必不可少的 了解细胞命运调节器。结合小鼠遗传学，纯化的散装和 单细胞、3D成像和人类干细胞技术，这一提议将检验我们的中心假设 NKX2-1和Wnt信号转录因子的顺序和组合作用控制着 肺上皮细胞类型在体内的表观遗传成熟和维持。该提案具有 遵循三个目标。(1)检测肺命运维持是否从WNT依赖向NKX2-1自身转移 随着肺的成熟而增强。(2)检测NKX2-1是否促进AT1细胞与AT2细胞作为 YAP/TAZ/TEAD与CEBPA的招募结果。(3)测试NKX2-1 为AT1分化准备AT2细胞。

PROJECT SUMMARY The lung epithelium consists of diverse cell types, as the result of embryonic progenitors undergoing a series of precise cell fate decisions. Upon injury, a successful repair needs to restore the proper cell fates. Recapitulation and modulation of in vivo cell fate biology holds promise for cell-based regeneration therapy. Despite such fundamental importance and our accumulating knowledge of cell fate regulators, the epigenetic basis of lung cell fate – defined in this proposal as chromatin state and its relationship to transcription factor binding – is largely unknown, but essential for mechanistic understanding of cell fate regulators. Combining mouse genetics, genomics of both purified bulk and single cells, 3D imaging, and human stem cell technology, this proposal will test our central hypothesis that sequential and combinatorial actions of NKX2-1 and Wnt signaling transcription factors control the epigenetic maturation and maintenance of lung epithelial cell types in vivo. The proposal has the following 3 aims. (1) To test whether lung fate maintenance shifts from Wnt-dependent to NKX2-1 self- reinforcing as the lung matures. (2) To test whether NKX2-1 promotes AT1 versus AT2 cell fate as a result of recruitment by YAP/TAZ/TEAD versus CEBPA, respectively. (3) To test whether NKX2-1 primes AT2 cells for AT1 differentiation.