Epithelial Progenitor Cells in Lung Repair and Regeneration

描述(申请人提供)：肺细支气管壁和肺泡上皮显著参与许多与肺泡完整性丧失相关的重要人类疾病，包括肺气肿和肺纤维化。在每一种情况下，产生新的肺泡上皮及其相关的血管床的能力都将具有巨大的潜在治疗价值，但这种能力仍然超出了当前技术的能力范围。这一应用的基础是这样一种想法，即更好地了解远端气道和肺泡上皮细胞前体细胞生物学是将定向DIGH肺重塑或修复领域推向临床的关键部分。为此，这项应用将在上皮祖细胞生物学方面具有不同专业知识但兴趣重叠的研究小组聚集在一起，以促进对远端肺的发育、维持和修复的理解。本研究的主要目的是(1)确定迄今尚未确定的远端气道和肺泡祖细胞的转录程序，并验证黏附受体的差异表达是上皮亚型自我组织和促进修复能力的基础这一假说。(2)明确肺损伤后对神经内分泌细胞(PNECs)和肺泡祖细胞在维持和重建远端气道和肺泡细胞中的需求。(3)为优化成人上皮祖细胞产生新的功能呼吸单位的能力，分析并进一步发展一种新的肾胶囊内单细胞活体肺器官测定方法。为实现这些目标而开发的重要工具和方法包括具有可诱导的cre活性的小鼠，敲入谱系定义的基因组基因座的小鼠，基于流式细胞术的分离和转录描述小鼠和人类胚胎和成人上皮祖细胞的技术，以及允许实时捕捉肺和肺器官随时间变化的稳定图像的创新成像。我们预计，在完成这些研究后，我们应该能够使我们的体内检测方法适用于具有肺发育能力的细胞单位的原位移植。总体而言，这些研究应该为从祖细胞生物学到人类肺部疾病的临床转化提供关键的概念和技术基础。相关性(见说明)；这项建议汇集了几名在上皮细胞生物学领域具有重叠但不同专业知识的研究人员，目标是产生概念性生物基础设施以及从祖细胞来源创建远端肺组织从头开始的技术。这些项目应该能够将新概念转化为肺部疾病的治疗。

DESCRIPTION (provided by applicant): The pulmonary bronchiolar and alveolar epithelia are involved prominently in a number of important human diseases associated with loss of alveolar integrity, including emphysema and pulmonary fibrosis. In each of these conditions the capacity to generate new alveolar epithelium, and its associated vascular bed, would be of great potential therapeutic value, but such capacity remains beyond the reach of current technology. This application is predicated on the idea that better understanding of distal airway and alveolar epithelial cell progenitor biology is a crucial part of any effort to move the field of directed dital lung remodeling or repair toward the clinical arena. Toward that end, this application brings together investigative groups with different expertise but overlapping interests in epithelial progenitor cell biology to advance the understanding of distal lung development, maintenance, and repair. The major objectives are (1) to define the transcriptional program of heretofore uncharacterized distal airway and alveolar progenitors and test the hypothesis that differential expression of adhesion receptors underlies the capacity of epithelial subtypes to self-organize and promote repair. (2) Define the requirement for neuroendocrine cells (PNECS) and alveolar progenitor cells in maintenance and reconstitution of distal airway and alveolar cells following lung injury. (3) Analyze and further develop a novel, single cell in vivo lung organoid assay in kidney capsules in order to optimize the capacity of adult epithelial progenitor cells to generate functional respiratory units de novo. Important tools and approaches developed to achieve these aims include mice with inducible cre activity knocked into lineage-defining genomic loci, flow cytometry-based techniques to isolate and transcriptionally profile mouse and human embryonic and adult epithelial progenitors, and innovative imaging that allow real time capture of stable images of lung and lung organoids over time. We anticipate that by the completion of these studies we should be able to adapt our in vivo assay toward orthotopic transplantation of cellular units capable of lung development. Overall, these studies should provide crucial conceptual and technological infrastructure for the clinical translation of progenitor cell biologyto human lung disease. RELEVANCE (See instructions); This proposal brings together several investigators with overlapping but distinct expertise in the field of epithelial cell biology with he goal of generating the conceptual biological infrastructure as well as technology for creating distal lung tissue de novo from a source of progenitor cells. The projects should empower translation of new concepts to the treatment of lung diseases.