Genetic and Molecular Dissection of Pulmonary Neuroendocrine (NE) Cell Development

项目摘要 NE细胞是肺中最有趣但知之甚少的细胞类型之一。许多分布在 在支气管上皮细胞中明显随机分布，但其他细胞被组织成20-30个NE细胞的簇，称为 神经上皮小体（NEB）是由传入和传出纤维高度支配的。它们分泌出一种 血管活性神经肽和胺，并在呼吸生理学中具有许多功能，包括 气道氧感测、机械感测、支气管张力的调节和肺血流的局部控制。 NE细胞也具有在肺损伤中激活的干细胞功能，并且是小细胞肺癌中的肿瘤起始细胞 小细胞肺癌（SCLC），一种高度转移和最致命的肺癌形式。NE细胞扩张或显示分布改变 然而，在许多呼吸系统疾病中，对它们的正常发育、增殖、位置 分化和生理功能。 该职业发展奖的候选人对NEB进行了全面的研究 在小鼠中的发展，揭示其形成的细胞机制，包括一种新的形式的靶向 上皮细胞迁移称为滑动。这种意想不到的上皮细胞重排模式可能解释了 小细胞肺癌的高度侵袭性和迁移性。该提案旨在应用单细胞全- 对发育中的小鼠NE细胞进行基因组转录组分析（单细胞RNAseq）以阐明完整基因 滑动的表达程序。将建立在发育中的NE细胞中进行条件性基因缺失的系统， 并用于确定在初步单细胞RNAseq研究中鉴定的候选滑动基因的功能。在 与此同时，这种对小鼠NE细胞发育的新认识将通过对人类NE细胞的检测而扩展到人类。 在发育中的NE细胞中滑动基因的表达。 研究的主要地点是在生物化学系，位于贝克曼中心， 分子和遗传医学，其中还设有发育生物学系和 分子和细胞生理学系，以及霍华德休斯医学研究所的其他部门。 这些部门是世界上在这些领域的主要部门之一，并一直处于领先地位。 几十年来，生物化学，分子生物学，遗传学和基因组学的创新中心。贝克曼 中心附属于医院，位于斯坦福大学医学院校园的中心， 从斯坦福大学主校区的街道。 候选人在小鼠中最先进的遗传策略方面的专业知识和对以下方面的丰富知识： 人类呼吸道疾病将补充目前的培训计划，学习单细胞转录组分析 通过与斯坦福大学创新技术领域的领导者合作， NE细胞的发育和疾病这项工作的长期目标是确定分子缺陷 与NE细胞发育异常相关的潜在儿科和成人呼吸道疾病， 多样性和功能，以设计有效的治疗方法。

PROJECT SUMMARY NE cells are among the most interesting, but poorly understood cell types in the lung. Many are distributed apparently randomly in the bronchial epithelium, but others are organized into clusters of 20-30 NE cells called neuroepithelial bodies (NEBs) that are highly innervated by afferent and efferent fibers. They secrete a variety of vasoactive neuropeptides and amines and have many proposed functions in respiratory physiology, including airway oxygen sensing, mechanosensing, regulation of bronchial tone, and local control of pulmonary blood flow. NE cells also have a stem-cell function activated in lung injury and are tumor-initiating cells in small cell lung cancer (SCLC), a highly metastatic and the most deadly form of lung cancer. NE cells expand or show altered distributions in many respiratory diseases, however, little is known about their normal development, proliferation, placement, differentiation and physiologic function. The candidate for this career development award has performed a comprehensive study of NEB development in mice, revealing the cellular mechanism of their formation, including a novel form of targeted epithelial cell migration called slithering. This unexpected mode of epithelial cell rearrangement may explain the highly invasive and migratory nature of small cell lung cancer. This proposal aims to apply single cell whole- genome transcriptome analysis (single cell RNAseq) to developing mouse NE cells to elucidate the complete gene expression program of slithering. A system for conditional gene deletion in developing NE cells will be established, and used to determine the function of a candidate slithering gene identified in preliminary single cell RNAseq studies. In parallel, this emerging understanding of mouse NE cell development will be extended to humans by testing for human expression of slithering genes in developing NE cells. The primary site of research is within the Biochemistry Department, located in the Beckman Center for Molecular and Genetic Medicine, which also houses the Department of Developmental Biology and the Department of Molecular and Cellular Physiology, as well as the rest of the Howard Hughes Medical Institute. These Departments are among the premier departments in the world in these fields, and have been leading centers of innovation in biochemistry, molecular biology, genetics, and genomics for decades. The Beckman Center is attached to the hospital and centrally located on the Stanford Medical School campus, and across the street from the main Stanford University campus. The candidate's expertise in the most advanced genetic strategies in mice and strong knowledge of human respiratory diseases will complement the current training plan to learn single cell transcriptome analysis through collaborations with leaders in this innovative technology at Stanford to address important questions about NE cell development and disease. The long term goal of this work is to identify the molecular defects underlying pediatric and adult respiratory diseases associated with abnormalities in NE cell development, diversity, and function(s) in order to design effective therapies.