Extracellular matrix composition and crosslinking patterns determine resident cell function in lung fibrosis

细胞外基质组成和交联模式决定肺纤维化中的常驻细胞功能

基本信息

批准号：
9887758
负责人：
Oliver Eickelberg
金额：
$ 66.19万
依托单位：
UNIVERSITY OF COLORADO DENVER
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：
2019-12-15 至 2020-07-21
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9887758
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 Airway Disease Chronic lung disease Cicatrix Complex Control Animal Cues Data Set Deposition Detergents Development Digestion Disease Enzymes Epithelial Epithelial Cells Epithelium 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 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 small hairpin RNA three-dimensional modeling tissue regeneration transcriptome sequencing transglutaminase 2

项目摘要

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.

项目摘要/摘要特发性肺纤维化（IPF）是一种进行性致命的慢性肺部疾病，影响超过500万人全世界。迄今为止，尚无有效阻止进展或扭转疾病的疗法。 IPF是特征是细胞组成改变和上皮 - 间质相互作用的功能障碍周围肺，导致细胞外基质（ECM）和进行性疤痕的过度积累。这 IPF肺的特征是正常或受影响区域的异质分布，与大量纤维化区域含有间隔增厚，蜂窝，异常上皮重编程，和成纤维细胞灶。由于稳态和受伤后人肺的再生受到精致的控制 ECM与多个居民细胞群之间的相互作用，必须定义顺序 ECM分泌增强和对细胞功能的交联的贡献。因此，定义通过在交联酶活性上获得的增强ECM组成或刚度的顺序层次结构居民肺部细胞功能将使IPF的精确治疗角度识别。总体该应用的目标是将纤维化ECM的组成和交联模式定义为评估成纤维细胞对纤维化ECM的贡献，以机械询问贡献上面的原型交联酶，转谷氨酰胺酶（TGM）2，并评估其倒数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提示，由居民成纤维细胞产生并由TGM2依赖性交联产生，进而改变肺上皮细胞功能和重编程。为了提出这一假设，我们提出了一系列特定目的的级联：在AIM1中，我们将利用一种新型的蛋白质组学方法来定义，量化和验证，以最大的可能细节和准确性，通过量化其肺纤维化中ECM的组成和结构的变化组成和交联模式。在AIM 2中，我们建议确定控制和IPF分泌的ECM 原代成纤维细胞并确定成纤维细胞衍生的TGM2对ECM组成和交联的影响。在AIM 3中，我们将研究成纤维细胞衍生的TGM2是否以及如何影响肺纤维化和 ATII细胞重编程。该提议基于一个新概念，即居民肺部命运是相互的由（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的人肺中的ECM组成和交联模式。我们将生成小说关于IPF中常驻肺部细胞功能和组织再生的ECM细胞相互作用的知识。这项目将探索肺部病理中一个主要不足的区域，并为 IPF新型疗法的开发可能会扩展到其他由其他慢性肺部疾病。 ECM组成的变化，例如哮喘，慢性肺同种异体功能障碍或COPD。