ECM Proteomics in lung fibrosis

项目摘要/摘要 特发性肺纤维化(Ipf)是一种进行性和致命性慢性肺部疾病，影响超过500万人。 全世界。到目前为止，还没有有效阻止病情发展或逆转疾病的治疗方法。IPF是 以细胞成分改变和上皮-间充质相互作用障碍为特征 周围肺，导致细胞外基质(ECM)过度堆积和进行性瘢痕形成。这个 IPF肺的特点是正常或轻度受影响的区域分布不均，与 显著的纤维化区域，包括间隔增厚、蜂窝状、异常的上皮重新编程， 和成纤维细胞病灶。由于人体肺损伤后的动态平衡和再生是由微妙的 ECM和多个驻留细胞群之间的相互作用，必须定义顺序 细胞外基质分泌增强和交联对细胞功能的贡献。因此，对 通过交联酶活性获得的增强的ECM组成或硬度的顺序层次 驻留肺细胞功能将使IPF的精确治疗角度得以确定。最重要的是 本申请的目的是定义纤维化ECM的组成和交联型式，以 评估成纤维细胞在纤维性ECM中的作用，以机械方式询问其作用 一种原型交联酶转谷氨酰胺酶(TGM)2对上述作用，并评估其相互作用 对肺泡上皮细胞功能的影响。我们假设IPF ECM显示出特定的变化和线索， 由常驻成纤维细胞产生，由TGM2依赖的交联物产生，进而改变肺上皮细胞 细胞功能和重新编程。为了推进这一假设，我们提出了一系列具体目标：在Aim1中， 我们将利用一种新的蛋白质组学方法来最大限度地定义、量化和验证 肺纤维化中细胞外基质成分和结构的变化 合成和交联型图案。在目标2中，我们建议鉴定对照和ipf分泌的细胞外基质。 并确定成纤维细胞来源的TGM2对细胞外基质组成和交联度的影响。 在目标3中，我们将调查成纤维细胞来源的TGM2是否以及如何影响肺纤维化的发展和 ATII细胞重新编程。这一建议是基于常驻肺细胞命运是相互的这一新概念。 由(纤维化的)ECM决定。拟议的项目将提供前所未有的细节和新颖的见解 正常人和纤维化人肺中细胞外基质的组成和交联型。我们将创作一部小说 关于细胞外基质-细胞相互作用与IPF中驻留肺细胞功能和组织再生的知识。这个 该项目将探索肺部病理研究不足的主要领域，并为 IPF新疗法的开发，可能会扩展到其他慢性肺部疾病，由 ECM成分的变化，如哮喘、慢性肺移植功能障碍或COPD。

PROJECT SUMMARY/ABSTRACT Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal chronic lung disease, affecting over 5 million people worldwide. To date, there are no therapies that effectively stop progression or reverse the disease. IPF is characterized by altered cellular composition and dysfunction of epithelial-mesenchymal interaction in the peripheral lung, leading to excessive accumulation of extracellular matrix (ECM) and progressive scarring. The IPF lung is characterized by a heterogeneous distribution of normal or mildly affected regions, alternating with regions of significant fibrosis containing septal thickening, honeycombing, aberrant epithelial reprogramming, and fibroblastic foci. Since homeostasis and regeneration of the human lung after injury is controlled by delicate interplay between the ECM and multiple resident cell populations, it is imperative to define the sequential contributions of enhanced ECM secretion and crosslinking on cellular functions. Hence, the definition of the sequential hierarchy of enhanced ECM composition or stiffness obtained by crosslinking enzyme activity on resident lung cell function will enable the identification of precise therapeutic angles for IPF. The overarching goal of this application is to define the composition and crosslinking pattern of the fibrotic ECM, to assess the contribution of fibroblasts to the fibrotic ECM, to mechanistically interrogate the contribution of a prototypic crosslinking enzyme, transglutaminase (TGM) 2, to the above, and to assess its reciprocal effect on alveolar epithelial cell function. We hypothesize that IPF ECM exhibits specific changes and cues, produced by resident fibroblasts and generated by TGM2-dependent crosslinks, which in turn alter lung epithelial cell function and reprogramming. To pursue this hypothesis, we propose a cascade of specific aims: In Aim1, we will utilize a novel proteomics approach in order to define, quantify, and validate, in the greatest possible detail and accuracy, changes in the composition and architecture of the ECM in lung fibrosis by quantifying its composition and crosslinking patterns. In Aim 2, we propose to identify the ECM secreted by control and IPF primary fibroblasts and determine the effect of fibroblast-derived TGM2 on ECM composition and crosslinking. In Aim 3, we will investigate whether and how fibroblast-derived TGM2 affects development of lung fibrosis and ATII cell reprogramming. This proposal is based on the new concept that resident lung cell fate is reciprocally determined by the (fibrotic) ECM. The proposed project will provide unprecedented detail and novel insights into ECM composition and crosslinking patterns in the normal and fibrotic human lung. We will generate novel knowledge on ECM-cell interaction with respect to resident lung cell function and tissue regeneration in IPF. The project will explore a major under-investigated area in lung pathologies and provide substantial groundwork for the development of novel therapies for IPF, which likely will extend to other chronic lung diseases driven by changes in ECM composition, such as asthma, chronic lung allograft dysfunction, or COPD.