Chymotrypsin-like Elastase 1 in Lung Development and Disease

项目摘要 正常的出生后肺形态发生和损伤后的修复需要精确协调 细胞活动和基质重塑对正常肺泡形成或恢复的作用 结构和最佳呼吸效率。我们最近确定了蛋白酶的一个关键作用， 凝乳酶样弹力酶1(CELA1)对出生后肺弹性的调节及其临界 在α-1抗胰蛋白酶缺乏症发病机制中的作用这项研究计划是基于 关于我们新的初步发现，Cela1在再生过程中被诱导表达 和病理性肺泡重塑，抑制Cela1活性可预防AAT 缺乏性肺气肿。这项建议将确定Cela1在生理学和生理学上的作用 以及出生后肺的病理重塑。根据Cela1表达式的数据 高氧性肺损伤中的调节失调，AIM 1将确定CELA1在肺中的细胞特异性作用 使用条件缺失模型鉴定表达Cela1的细胞和 正常和高氧条件下发育过程中基因表达的动态变化 条件。目的：检测Cella1在病理性肺组织中的细胞特异性表达。 基质重塑和靶向Cela1是否能长期预防肺气肿。vbl.使用 AAT缺陷性肺气肿模型的谱系追踪和邻近连接原位杂交， 我们将确定表达Cela1的ATII细胞是否代表一种独特的上皮细胞亚类 或者如果所有的ATII细胞都可以在适当的上下文中表达Cela1。在AIM 3中，AAT是否 细胞摄取所需的CELA1的中和作用将被测试，参与这一过程的细胞 AAT-Cela1复合体的摄取和再循环将通过谱系追踪、邻近 连接原位杂交、共聚焦显微镜和流式细胞术。它们之间的联系 AAT缺陷者的Cela1基因表达和弹性蛋白重塑区域将被确定 肺气肿标本。最后，确定了CELA1-AAT分子相互作用的位置。从一个 从科学和临床的角度来看，这项提议将定义一种新的关键机制，通过 CELA1介导基质重塑过程调节正常牙槽骨形成和再生 在受伤后，这将为探索Cela1作为发展的靶点提供强有力的理由 间质性肺病的未来治疗方法。

Project Summary Normal postnatal lung morphogenesis and repair after injury requires the precise coordination of cellular activities and matrix remodeling for the formation or restoration of normal alveolar structures and optimal respiratory efficiency. We recently identified a critical role for the protease, Chymotrypsin-like elastase 1 (Cela1), in the regulation of postnatal lung elastance and its critical role in the pathogenesis of α1-antitrypsin (AAT) deficiency. This program of research is based on our novel preliminary findings that Cela1 expression is induced during regenerative and pathological alveolar remodeling, and that inhibition of Cela1 activity prevents AAT deficiency related emphysema. This proposal will identify the role of Cela1 in both physiological and pathologic remodeling in the postnatal lung. Based on data that Cela1 expression is dysregulated in hyperoxic lung injury, Aim 1 will define cell-specific roles for Cela1 in lung development using conditional deletion models to identify Cela1-expressing cells and characterize dynamic changes in expression during development under normal and hyperoxic conditions. Aim 2 will test whether cell-specific expression of Cela1 changes in pathological lung matrix remodeling and whether targeting Cela1 can protect against emphysema long-term. Using lineage tracing and proximity ligation in situ hybridization in an AAT-deficient emphysema model, we will determine whether Cela1-expressing ATII cells represent a unique epithelial cell subclass or if all ATII cells can express Cela1 in the appropriate context. In Aim 3, whether AAT neutralization of Cela1 is required for cellular uptake will be tested, and the cells involved in this uptake and recycling of AAT-Cela1 complexes will be identified using lineage tracing, proximity ligation in situ hybridization, confocal microscopy, and flow cytometry. The association among Cela1 gene expression and regions of elastin remodeling will be identified in human AAT deficient emphysema specimens. Lastly, site of Cela1-AAT molecular interaction will be defined. From a scientific and clinical standpoint, this proposal will define a novel critical mechanisms by which Cela1 mediates matrix remodeling processes to regulate normal alveolarization and regeneration after injury that will provide a strong rationale to explore Cela1 as a target for development of future therapies for interstitial lung diseases.