Epithelial Progenitor Cells in Lung Repair and Regeneration

描述（由申请人提供）：肺细支气管和肺泡上皮细胞在许多与肺泡完整性丧失相关的重要人类疾病中有显著的参与，包括肺气肿和肺纤维化。在这些病症中的每一种中，产生新肺泡上皮及其相关血管床的能力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范围。这种应用基于这样的想法，即更好地理解远端气道和肺泡上皮细胞祖细胞生物学是将定向远端肺重塑或修复领域推向临床竞技场的任何努力的关键部分。为此，该应用程序汇集了在上皮祖细胞生物学方面具有不同专业知识但重叠兴趣的研究小组，以促进对远端肺发育，维护和修复的理解。主要目的是（1）确定迄今为止未表征的远端气道和肺泡祖细胞的转录程序，并测试粘附受体的差异表达是上皮亚型自组织和促进修复的能力的基础的假设。(2)明确肺损伤后维持和重建远端气道和肺泡细胞对神经内分泌细胞（PNECS）和肺泡祖细胞的需求。(3)分析并进一步开发一种新的肾包膜中的单细胞体内肺类器官测定，以优化成体上皮祖细胞从头产生功能性呼吸单位的能力。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cre活性的小鼠敲入谱系定义基因组位点，基于流式细胞术的技术来分离和转录分析小鼠和人胚胎和成人上皮祖细胞，以及创新成像，其允许随时间推移真实的时间捕获肺和肺类器官的稳定图像。我们预计，通过完成这些研究，我们应该能够调整我们的体内试验对原位移植的细胞单位能够肺的发展。总的来说，这些研究应该为人类肺部疾病的祖细胞生物学临床翻译提供关键的概念和技术基础设施。相关性（参见说明）;该提案汇集了几位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，其目标是产生概念性生物基础设施以及从祖细胞来源从头创建远端肺组织的技术。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.