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Interferon-mediated in vitro and in vivo Regulation of Airway Progenitor Cells in Regeneration of Functional Alveoli

干扰素介导的气道祖细胞在功能性肺泡再生中的体外和体内调节

基本信息

批准号：
9925055
负责人：
Jaymin J Kathiriya
金额：
$ 5.35万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2021-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9925055
关键词：
Acute Adult Alveolar Biology Bleomycin CDKN1C gene Cell Cycle Cell Therapy Cells Chronic lung disease Cicatrix Cyclins Cytokeratin Data Development Disease Distal Embryo Engraftment Epithelial Epithelial Cells Epithelium Family Fibrosis Foundations Future Gene Expression Genes Goals In Vitro Injury Interferon Type II Interferon-alpha Interferons Knowledge Label Link Location Lung Lung Capacity Maintenance Major Histocompatibility Complex Gene Measures Mediating Minor Mus Natural regeneration Organoids Outcome Patients Population Process Proliferating Proteins Pulmonary Surfactant-Associated Protein C Recovery Recovery of Function Regulation Resolution Respiratory physiology Role STAT1 gene Signal Pathway Signal Transduction Source Suggestion Testing Therapeutic Agents Transplantation Work airway epithelium alveolar epithelium alveolar type II cell computerized tools epithelial stem cell epithelium regeneration experimental study genetic signature genomic platform in vivo inhibitor/antagonist injured injury and repair insight lung injury lung regeneration member mouse model novel novel therapeutics progenitor pulmonary function receptor reconstitution regenerative repaired response response to injury severe injury single cell analysis single cell sequencing stem stem cells surfactant transcriptome transcriptomics virtual

项目摘要

PROJECT SUMMARY Recapitulation of normal lung function following a severe acute injury implies an inherent capability of lung to regenerate but the sources and relative regenerative capacities of lung epithelial stem/progenitor cells remain unclear. Although virtually all differentiated epithelial lineages can re-enter the cell cycle, increasing evidence indicates that subpopulations of distal airway cells may be particularly robust in their capacity to expand, migrate, and reconstitute alveolar barriers. Recent studies from our lab have uncovered Sox2pos/p63neg epithelial progenitors (EPs) with regenerative potential and ability to differentiate towards alveolar fate. Further, numerous other studies have described a fraction of Scgb1a1-labeled cells to also have in vitro and in vivo regenerative capacity. However, their exact identities, location, and regulation remains unclear. To further define the EPs, we performed large scale single cell sequencing of flow-sorted lung epithelial cells enriched in EPs, which identified a quiescent EP population. These cells, with a transcriptomic similarity to embryonic Sox9pos lung bud cells, have increased expression of interferon (IFN)-regulated genes. Interestingly, these cells also express a number of genes associated with cell cycle and key cyclin inhibitors Cdkn1a (p21) and Cdkn1c (p57), suggestive of their ability to proliferate under favorable conditions. Indeed, IFNpos cells represent the regenerative fraction of the airway epithelium (including that of Scgb1a1-labeled cells) in various in vitro conditions tested. Importantly, these cells also have the ability to differentiate towards either basal (marked by Cytokeratin 5 expression) or alveolar (marked by surfactant protein C expression) cells. Thus, my preliminary data have identified the regenerative fraction of the airway epithelium. However, their regulation in vivo and their ability to functionally contribute to lung epithelial regeneration post injury remain unknown. To this end, we hypothesize that low levels of constitutive IFN signaling regulates and maintains a pool of rare airway EPs that mobilize to regenerate alveolar epithelium following injury. We will test this hypothesis in following two aims: 1) To determine the role of IFN signaling in maintenance of quiescent p63neg EPs in uninjured mouse lung. 2) Determine the role of IFN signaling in in vivo responses of p63neg EPs following major injury. We will leverage mouse models to abrogate active IFN signaling in injured mice to study its role in maintenance of EPs. Likewise, conditional airway epithelial deletion of Stat1 followed by injury will determine the role of IFN/Stat1 signaling in injury response. Finally, exogenously expanded EPs will be transplanted in injured lungs and functional recovery will be measured by oxygenation and lung function. This project has a significant potential to clarify the signaling pathways regulating lung epithelial progenitor cell response during quiescence and determine the potential of these epithelial progenitors in aiding lung regeneration following an injury. Further, the novel studies involving transplantation of exogenously expanded progenitors will lay foundation for their overall regenerative potential as a therapeutic agent for future cell based therapies.

项目摘要严重急性损伤后正常肺功能的重现意味着肺的固有能力，但肺上皮干/祖细胞的来源和相对再生能力仍然存在不清楚尽管几乎所有分化的上皮细胞都可以重新进入细胞周期，但越来越多的证据表明，表明远端气道细胞的亚群在它们的扩张，迁移，重建肺泡屏障我们实验室最近的研究发现，Sox2pos/p63 neg上皮细胞具有再生潜力和向肺泡命运分化的能力的祖细胞（EP）。此外，许多其他研究已经描述了Scgb1a1标记的细胞的一部分也具有体外和体内再生能力，容量然而，它们的确切身份、位置和监管仍不清楚。为了进一步定义EP，我们对富集EP的流式分选肺上皮细胞进行大规模单细胞测序，一个安静的EP人口。这些细胞与胚胎Sox9pos肺芽细胞具有转录组相似性，干扰素（IFN）调节基因的表达增加。有趣的是，这些细胞也表达一些与细胞周期和关键细胞周期蛋白抑制剂Cdkn1a（p21）和Cdkn1c（p57）相关的基因，提示它们可能与细胞周期相关。在有利条件下增殖的能力。事实上，IFNpos细胞代表了肿瘤细胞的再生部分。气道上皮细胞（包括Scgb1a1标记细胞）在各种体外条件下的测试。重要的是这些细胞也具有向基底（以细胞角蛋白5表达为标志）或肺泡分化的能力（以表面活性蛋白C表达为标志）细胞。因此，我的初步数据已经确定了再生气道上皮的一部分。然而，它们在体内的调节和它们在功能上有助于损伤后肺上皮再生仍然未知。为此，我们假设低水平的组成性IFN信号调节并维持一组罕见的气道EP，损伤后的肺泡上皮。我们将在以下两个目标中检验这一假设：1）确定 IFN信号在未损伤小鼠肺中维持静止p63 neg EP中的作用。2)确定的作用严重损伤后p63 neg EP体内应答中的IFN信号传导。我们将利用小鼠模型，在受损小鼠中消除活性IFN信号传导以研究其在维持EP中的作用。同样，条件气道损伤后上皮细胞Stat1的缺失将决定IFN/Stat1信号传导在损伤应答中的作用。最后，将外源性扩增的EP移植到受损的肺中，并测量功能恢复通过氧合和肺功能。这个项目有一个重要的潜力，以澄清信号通路在静止期调节肺上皮祖细胞反应，并确定这些细胞的潜力，上皮祖细胞在损伤后帮助肺再生中的作用。此外，新的研究涉及移植外源扩增的祖细胞将为其整体再生潜力奠定基础作为未来细胞疗法的治疗剂。