Transcriptional and epigenetic basis of lung epithelial cell fate

项目摘要 肺上皮细胞由不同类型的细胞组成，这是胚胎祖细胞经历一个 一系列精确的细胞命运决定。一旦受伤，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重演和调节为基于细胞的免疫调节提供了希望。 再生疗法尽管有这样的基本重要性和我们积累的知识，细胞 命运调节因子，肺细胞命运的表观遗传基础-在该提案中定义为染色质状态， 它与转录因子结合的关系-在很大程度上是未知的，但对于 了解细胞命运调节器。结合小鼠遗传学、纯化的原液和 单细胞，3D成像和人类干细胞技术，这项提案将测试我们的中心假设 NKX 2 -1和Wnt信号传导转录因子的顺序和组合作用控制了细胞的增殖， 表观遗传成熟和体内肺上皮细胞类型的维持。该提案具有 三个目标。(1)为了测试肺命运维持是否从Wnt依赖性转变为NKX 2 -1自身依赖性， 随着肺的成熟而增强。(2)为了测试NKX 2 -1是否促进AT 1相对于AT 2细胞的命运， 分别由雅普/TAZ/TEAD与CEBPA招募的结果。(3)为了测试NKX 2 -1是否 为AT 1分化准备AT 2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.