Defining and targeting mechanisms of smoke-mediated fibrosis progression

烟雾介导的纤维化进展的定义和靶向机制

基本信息

批准号：
10586073
负责人：
Ching-Hsien Chen
金额：
$ 39.25万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
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：
2019-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10586073
关键词：
Achievement Actins Adoptive Transfer Affect Antibodies Binding Biological Assay Biological Markers Biopsy Bleomycin Cell Proliferation Cells Chronic lung disease Co-Immunoprecipitations Complement Complex Data Development Disease Down-Regulation Environmental Exposure Exposure to FDA approved Fibroblasts Fibrosis Future Gene Expression Profiling Genes Human Immunofluorescence Immunologic In Vitro Interstitial Lung Diseases Intervention Knock-out Ligands Lung MARCKS gene Mainstreaming Malignant Neoplasms Malignant neoplasm of lung Mediating Medical Membrane Proteins Modeling Molecular Myofibroblast Pathogenesis Pathogenicity Patients Peptides Pharmaceutical Preparations Phenotype Phosphorylation Phosphotransferases Play Prognosis Proteins Pulmonary Fibrosis Receptor Protein-Tyrosine Kinases Reporting Risk Role Series Signal Transduction Signaling Protein Smoke Smoking Smoking History Smoking Status Smooth Muscle Structure of parenchyma of lung Testing Therapeutic Time Tissues Tobacco smoke Tobacco smoking behavior Tyrosine Phosphorylation Up-Regulation Western Blotting axl receptor tyrosine kinase cigarette smoke cigarette smoking clinically relevant druggable target epidemiology study evidence base exposed human population exposure to cigarette smoke fibrotic lung former smoker idiopathic pulmonary fibrosis in vivo inhibitor knock-down lung cancer cell migration new therapeutic target nintedanib novel pharmacologic response small hairpin RNA therapeutic target tumor progression

项目摘要

Several environmental exposures, especially tobacco smoking, have been associated with increased risk for idiopathic pulmonary fibrosis (IPF), a progressive and incurable interstitial lung disease. This disease has a median survival time of 3 to 5 years, worse than many cancers. With 41%-83% of IPF patients being current or former smokers, cigarette smoking plays a crucial role in IPF development and progression. Unfortunately, the mechanisms by which smoking promotes fibrosis are poorly understood. Of great concern, there are no reliable biomarkers and/or specific therapies available for smoking-associated IPF. To discover the regulators of IPF fibroblast activation, we compared and integrated the gene profiles of IPF fibroblasts from two different microarray platforms. Among the genes identified, myristoylated alanine-rich C-kinase substrate (MARCKS) was found to be highly correlated with the expression of the myofibroblast marker alpha smooth muscle actin (α-SMA) and was notably elevated in IPF fibroblasts. We previously identified the membrane-associated protein MARCKS as a smoke-responsive molecule associated with lung cancer progression. Our preliminary data have demonstrated that up-regulation of phospho-MARCKS concomitant with an increase of protein tyrosine phosphorylation level in smoke-exposed cells. Using a phospho-kinase antibody array screen, we found the AXL receptor tyrosine kinase (RTK) as a top-one RTK active in response to smoke. In addition, our LC-MS/MS data have revealed AXL is a putative binding partner of phospho-MARCKS. Co-expression of phospho-MARCKS and phospho-AXL were observed in lung tissues with exposure to both mainstream and sidestream cigarette smoke. Surprisingly, the inhibition of phospho-MARCKS resulted in downregulation of AXL autophosphorylation and its downstream signaling. Given these observations, we hypothesize that cigarette smoke through MARCKS-AXL signaling stimulates fibroblast activation, thereby contributing to lung fibrosis progression. To test this hypothesis, three specific aims are proposed: 1) characterize the role of cigarette smoke-activated MARCKS-AXL axis in fibroblast cell proliferation, migration, and/or differentiation in vitro; 2) to determine that MARCKS inhibition effectively suppresses smoke-mediated lung fibrosis in vivo; 3) to evaluate the clinical relevance of phospho- MARCKS and phospho-AXL in smoking-related IPF tissues. Achievement of these specific aims will characterize novel molecular mechanisms underlying smoke-mediated fibrosis progression, thereby providing novel therapeutic targets such as phospho-MARCKS and phospho-AXL for combating pulmonary fibrosis.

多种环境暴露，尤其是吸烟，与罹患癌症的风险增加有关特发性肺纤维化（IPF），一种进行性且无法治愈的间质性肺疾病。中位生存时间为 3 至 5 年，比许多癌症更糟糕，其中 41%-83% 的 IPF 患者处于或处于状态。不幸的是，对于以前吸烟者来说，吸烟在 IPF 的发生和进展中起着至关重要的作用。人们对吸烟促进纤维化的机制知之甚少，也没有可靠的证据。可用于与吸烟相关的 IPF 的生物标志物和/或特定疗法发现 IPF 的调节因子。为了成纤维细胞活化，我们比较并整合了来自两种不同的 IPF 成纤维细胞的基因谱在已鉴定的基因中，富含肉豆蔻酰化的丙氨酸 C 激酶底物 (MARCKS) 是发现与肌成纤维细胞标记物α平滑肌肌动蛋白（α-SMA）的表达高度相关我们之前鉴定出膜相关蛋白 MARCKS 在 IPF 成纤维细胞中显着升高。作为与肺癌进展相关的烟雾反应分子。证明磷酸-MARCKS 的上调伴随着蛋白质酪氨酸的增加使用磷酸激酶抗体阵列筛选，我们发现了 AXL 暴露于烟雾的细胞中的磷酸化水平。受体酪氨酸激酶 (RTK) 作为对烟雾反应的首要 RTK 活性此外，我们的 LC-MS/MS 数据。已经揭示 AXL 是磷酸-MARCKS 的假定结合伙伴，并且磷酸-MARCKS 和的共表达。在暴露于主流和侧流香烟烟雾的肺组织中观察到磷酸-AXL。令人惊讶的是，磷酸-MARCKS 的抑制导致 AXL 自磷酸化及其鉴于这些观察结果，我们通过 MARCKS-AXL 追踪香烟烟雾。信号传导刺激成纤维细胞活化，从而促进肺纤维化进展。提出了三个具体目标：1）表征香烟烟雾激活的 MARCKS-AXL 轴在 2) 体外成纤维细胞增殖、迁移和/或分化，以确定MARCKS抑制；有效抑制体内烟雾介导的肺纤维化；3) 评估磷酸化的临床相关性与吸烟相关的 IPF 组织中的 MARCKS 和磷酸化 AXL 是这些特定目标的实现的特征。烟雾介导的纤维化进展的新分子机制，从而提供新的用于对抗肺纤维化的治疗靶点，例如磷酸-MARCKS 和磷酸-AXL。