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

项目总结 NE细胞是肺中最有趣但知之甚少的细胞类型之一。许多都是分布式的 表面上看是随机分布在支气管上皮中，但其他的则被组织成由20-30个NE细胞组成的簇，称为 由传入和传出纤维高度支配的神经上皮体。它们分泌各种各样的 血管活性神经肽和胺，在呼吸生理学中有许多拟议的功能，包括 呼吸道氧气感应、机械感应、调节支气管张力和局部控制肺血流。 NE细胞也具有在肺损伤中激活的干细胞功能，并且在小细胞肺癌中是肿瘤起始细胞 (SCLC)，一种高度转移性和最致命的肺癌形式。NE细胞扩张或分布改变 然而，在许多呼吸系统疾病中，人们对它们的正常发育、增殖、位置、 分化和生理功能。 这一职业发展奖的候选人对NEB进行了全面的研究 小鼠的发育，揭示了它们形成的细胞机制，包括一种新的靶向形式 上皮细胞的迁移称为滑动。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。这项提案旨在将单个细胞整体应用于- 对发育中的小鼠NE细胞进行基因组转录组分析(单细胞RNAseq)以阐明其完整基因 滑行的表达程序。建立NE细胞发育中的条件性基因缺失制度， 并用于确定初步单细胞RNAseq研究中确定的候选滑动基因的功能。在……里面 同时，这种对小鼠NE细胞发育的新兴理解将通过测试人类来扩展到人类 滑行基因在发育中的NE细胞中的表达 研究的主要地点在生物化学系内，位于贝克曼中心 分子和遗传医学，它也是发育生物学系和 分子和细胞生理学系，以及霍华德·休斯医学院的其他部门。 这些部门是世界上这些领域的主要部门之一，并一直处于领先地位 几十年来一直是生物化学、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，位于斯坦福医学院校园的中心，横跨 斯坦福大学主校区的街道。 候选人在最先进的小鼠遗传策略方面的专业知识和深厚的知识 人类呼吸道疾病将补充目前学习单细胞转录组分析的培训计划 通过与斯坦福大学这项创新技术的领导者合作，解决重要问题 关于NE细胞的发育和疾病。这项工作的长期目标是识别分子缺陷。 与NE细胞发育异常有关的儿童和成人呼吸系统疾病 多样性和功能性(S)，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.