ECM Proteomics in lung fibrosis

肺纤维化中的 ECM 蛋白质组学

• 批准号: 10352475
• 负责人: Oliver Eickelberg
• 金额: $ 61.67万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-15 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10352475
• 关键词: 3-Dimensional; Address; Affect; Architecture; Area; Asthma; Atlases; Biological Models; Bleomycin; Cell Communication; Cell Differentiation process; Cell physiology; Cells; Characteristics; Chemicals; Chronic; Chronic Obstructive Pulmonary Disease; Chronic lung disease; Cicatrix; Complex; Control Animal; Cues; Data Set; Deposition; Detergents; Development; Digestion; Disease; Enzymes; Epithelial; Epithelial Cells; Exhibits; Extracellular Matrix; Extracellular Matrix Proteins; Family; Fibroblasts; Fibrosis; Functional disorder; Genes; Goals; Histologic; Homeostasis; Human; Impairment; Injury; Interstitial Lung Diseases; Knock-out; Knowledge; Label; Lesion; Lung; Maps; Mass Spectrum Analysis; Mediating; Mesenchymal; Methods; Modeling; Molecular; Molecular Conformation; Mus; Natural regeneration; Normal Statistical Distribution; Pattern; Peripheral; Persons; Pharmacology; Population; Proteins; Proteome; Proteomics; Pulmonary Fibrosis; Pulmonary Pathology; Pulmonology; Site; Solubility; Structure; Structure of parenchyma of lung; Technology; Testing; Therapeutic; Transglutaminases; Type II Epithelial Receptor Cell; alveolar epithelium; base; cell type; crosslink; defined contribution; enzyme activity; genetic approach; idiopathic pulmonary fibrosis; improved; in vivo; injured; insight; knock-down; lung allograft; lung development; lung regeneration; man; next generation; novel; novel therapeutics; regenerative approach; small hairpin RNA; three-dimensional modeling; tissue regeneration; transcriptome sequencing; transglutaminase 2

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.

项目总结/摘要 特发性肺纤维化（IPF）是一种进行性和致死性慢性肺部疾病，影响超过500万人 国际吧到目前为止，还没有有效阻止疾病进展或逆转疾病的治疗方法。IPF是 其特征在于改变的细胞组成和上皮间质相互作用的功能障碍， 外周肺，导致细胞外基质（ECM）的过度积累和进行性瘢痕形成。的 IPF肺的特征是正常或轻度受累区域的不均匀分布，与 显著纤维化区域，包括间隔增厚、蜂窝样变、异常上皮重编程， 和成纤维细胞灶。由于损伤后人肺的内环境稳定和再生是由微妙的 ECM和多个驻留细胞群体之间的相互作用，必须定义序列 增强的ECM分泌和交联对细胞功能的贡献。因此， 通过交联酶活性获得的增强的ECM组合物或硬度的顺序层次， 常驻肺细胞功能将能够识别IPF的精确治疗角度。总体 本申请的目的是确定纤维化ECM的组成和交联模式， 评估成纤维细胞对纤维化ECM的贡献， 的原型交联酶，转氨酶（TGM）2，以上述，并评估其互惠 对肺泡上皮细胞功能的影响。我们假设IPF ECM表现出特定的变化和线索， 由常驻成纤维细胞产生并由TGM 2依赖性交联产生，这反过来改变了肺上皮细胞 细胞功能和重编程。为了实现这一假设，我们提出了一系列具体目标：在目标1中， 我们将利用一种新的蛋白质组学方法，以最大可能地定义、量化和验证 详细和准确性，通过定量肺纤维化中ECM的组成和结构的变化， 组成和交联模式。在目标2中，我们提出鉴定对照和IPF分泌的ECM 原代成纤维细胞，并确定成纤维细胞衍生的TGM 2对ECM组成和交联的影响。 在目的3中，我们将研究成纤维细胞来源的TGM 2是否以及如何影响肺纤维化的发展， ATII细胞重编程。这项建议是基于新的概念，即居民肺细胞的命运是不确定的。 由（纤维化）ECM确定。拟议的项目将提供前所未有的细节和新颖的见解， 正常和纤维化人肺中的ECM组成和交联模式。我们将创造新的 关于IPF中常驻肺细胞功能和组织再生的ECM-细胞相互作用的知识。的 该项目将探索肺部病理学的一个主要研究不足的领域，并为以下方面提供实质性的基础： IPF新型疗法的开发，可能会扩展到其他慢性肺部疾病， ECM组成的变化，如哮喘、慢性肺移植物功能障碍或COPD。