Collagen Oxidation, Myofibroblast Activation and Age-Associated Pulmonary Fibrosis

胶原蛋白氧化、肌成纤维细胞激活和年龄相关性肺纤维化

基本信息

批准号：
10532853
负责人：
Yvonne M. W. Janssen-Heininger
金额：
$ 48.29万
依托单位：
UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10532853
关键词：
Address Affect Age Aging Attenuated Autophagocytosis Bleomycin C-terminal Calnexin Cells Characteristics Cicatrix Collagen Cysteine Data Disease Environment Enzymes Extracellular Matrix Fibroblasts Fibrosis Glutathione Human Hydrogen Peroxide Intervention Knock-out Laboratories Link Lung Modeling Monitor Mus Myofibroblast Nature Oxidation-Reduction Pathogenesis Pathway interactions Patients Preparation Process Production Progressive Disease Proliferating Protein Disulfide Isomerase Proteins Proteomics Pulmonary Fibrosis Quality Control Recombinants Resistance Role Signal Transduction Solubility Stimulus Stress System TGFB1 gene Testing Tissues aged arm collagenase design effective therapy endoplasmic reticulum stress fibrotic lung glutaredoxin idiopathic pulmonary fibrosis improved indexing insight novel oxidation pulmonary function repair enzyme response trafficking

项目摘要

PROJECT SUMMARY Idiopathic pulmonary fibrosis (IPF) is a devastating, progressive aging-associated disease characterized by scarred lung tissue containing excessive extracellular matrix (ECM). Changes in the oxidative (redox) environment have long been known to accompany IPF, yet the mechanisms whereby redox perturbations affect IPF pathogenesis remain incompletely understood. Our laboratory has discovered that a specific type of protein oxidation, termed protein glutathionylation, is increased in the lungs of IPF patients and inversely correlates with lung function. Importantly, the activity of glutaredoxin (GLRX), an enzyme that reverses protein glutathionylation, was strongly decreased in IPF patient lungs. New studies from our laboratory showed that collagen 1A1 (COL1), a major component of the fibrotic ECM, is a target for glutathionylation (COL1-SSG) that is increased in IPF. Glutathionylation of COL1-SSG in the C-terminal pro-domain caused partial resistance to degradation by multiple collagenases. We also discovered that COL1-SSG is a potent activator of fibroblasts and causes oxidative signals that promote further glutathionylation. Collagens are the most abundant proteins produced in the ER and are challenging to properly assemble and process, requiring oxidative processes. Collagen has its own autophagy system consisting of calnexin (CANX) and FAM134B to remove aberrantly processed collagen via ER-linked autophagy. Autophagy is decreased in aging, and both CANX and FAM134B are decreased in lungs from IPF patients. These collective observations led us to hypothesize that in the aging lung, increases in ER oxidation and diminished collagen autophagy result in the secretion of COL1-SSG to promote the progression of pulmonary fibrosis by the activation myofibroblasts via a self-propagating stimulus that can be diminished by GLRX. In Specific Aim 1 we will address whether the activity of protein disulfide isomerase A3 which is important in the oxidative folding of COL1, and attendant increases in ER oxidoreductin 1-derived hydrogen peroxide contribute to COL1-SSG and the increases in age-associated persistent fibrosis. In Specific Aim 2 we will examine whether COL1-SSG is secreted by fibroblasts from fibrotic lung and whether this is linked to alterations in ER redox stress and collagen autophagy. In Specific Aim 3 we will elucidate whether GLRX status affects COL1-SSG, myofibroblast activation and age-associated lung fibrosis. Completion of these proposed studies that utilize a combination of targeted knockout models in aged mice and isolated (myo)fibroblasts from patients with IPF will provide novel insights into the mechanisms by which ECM dysregulation via oxidation (specifically glutathionylation) affects COL1 stiffness and promotes the progression of pulmonary fibrosis. Completion of the workplan will begin to unravel the potential role of aberrant collagen autophagy in IPF. Finally, completion of the proposed studies will also further elucidate the mechanisms of action whereby GLRX exerts anti-fibrotic activity and identify the potential improved anti-fibrotic action of a newly created oxidation-resistant version of GLRX.

项目概要特发性肺纤维化（IPF）是一种破坏性的、进行性的与衰老相关的疾病，其特征是疤痕肺组织含有过量的细胞外基质（ECM）。氧化（氧化还原）的变化人们早就知道环境会伴随 IPF，但氧化还原扰动影响的机制 IPF 的发病机制仍不完全清楚。我们的实验室发现了一种特定类型的蛋白质 IPF 患者肺部的氧化（称为蛋白质谷胱甘肽化）增加，并与肺功能。重要的是，谷氧还蛋白（GLRX）的活性，一种逆转蛋白质谷胱甘肽化的酶， IPF 患者肺部显着下降。我们实验室的新研究表明胶原蛋白 1A1 (COL1)、纤维化 ECM 的主要成分，是 IPF 中谷胱甘肽化 (COL1-SSG) 增加的靶标。 C 端前结构域中 COL1-SSG 的谷胱甘肽化导致对多种降解的部分抵抗胶原酶。我们还发现 COL1-SSG 是成纤维细胞的有效激活剂，可引起氧化促进进一步谷胱甘肽化的信号。胶原蛋白是内质网和内质网中产生的最丰富的蛋白质正确组装和加工具有挑战性，需要氧化过程。胶原蛋白有它自己的由钙联蛋白 (CANX) 和 FAM134B 组成的自噬系统，可通过去除异常加工的胶原蛋白 ER 相关的自噬。衰老过程中自噬减少，肺部 CANX 和 FAM134B 均减少来自 IPF 患者。这些集体观察结果使我们推测，在老化的肺部中，ER 增加氧化和胶原自噬减少导致 COL1-SSG 分泌，促进进展通过自我传播刺激激活肌成纤维细胞来抑制肺纤维化，这种刺激可以通过以下方式减少： GLRX。在具体目标 1 中，我们将讨论蛋白质二硫键异构酶 A3 的活性是否重要。 COL1 的氧化折叠，以及随之而来的 ER 氧化还原素 1 衍生的过氧化氢的增加导致 COL1-SSG 和与年龄相关的持续性纤维化的增加。在具体目标 2 中，我们将检查 COL1-SSG 是否由纤维化肺的成纤维细胞分泌，以及这是否与改变有关内质网氧化还原应激和胶原蛋白自噬。在特定目标 3 中，我们将阐明 GLRX 状态是否会影响 COL1-SSG，肌成纤维细胞活化和年龄相关的肺纤维化。完成这些拟议的研究利用老年小鼠的靶向敲除模型和患者分离的（肌）成纤维细胞的组合与 IPF 的合作将为 ECM 通过氧化（特别是谷胱甘肽）影响 COL1 硬度并促进肺纤维化的进展。完成工作计划将开始揭示异常胶原自噬在 IPF 中的潜在作用。最后，完成了拟议的研究还将进一步阐明 GLRX 发挥抗纤维化活性的作用机制并确定新创建的抗氧化版本 GLRX 潜在的改进抗纤维化作用。