Dedicator of cytokinesis 2 in pleural fibrosis

胸膜纤维化中胞质分裂2的奉献者

基本信息

批准号：
10683791
负责人：
Guoqing Qian
金额：
$ 36.28万
依托单位：
UNIVERSITY OF TEXAS HLTH CTR AT TYLER
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：
2022-09-15 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10683791
关键词：
Adipose tissue Asbestos Bleomycin Blood Coagulation Factor Blood Vessels Carbon Black Cells Cerebral Decortication Chest Cicatrix Clinical Collagen Cytokinesis Cytoskeletal Modeling Data Development Diet Disease Disease Progression Dyspnea Empyema Epithelial Event F-Actin Fatty acid glycerol esters Fibronectins Fibrosis Fructose Functional disorder Gene Expression Genes Goals Guanine Nucleotide Exchange Factors Hematopoietic Human Impairment Infection Inflammatory Intervention Knock-out Knockout Mice Knowledge Lead Lung diseases Manuscripts Mediating Mediator of activation protein Mesenchymal Mesothelial Cell Metabolic Modeling Molecular Morphology Mus Operative Surgical Procedures Outcome Pathogenesis Pathologic Processes Pathway interactions Patients Pharmacological Treatment Pharmacotherapy Phosphorylation Physiological Play Pleura Pleural Pleural Diseases Pleural Mesothelial Cell Pleurisy Pneumonia Process Public Health Pulmonary Inflammation Refractory Research Role Signal Pathway Signal Transduction Smooth Muscle Actin Staining Method Smooth Muscle Myocytes Snails Streptococcus pneumoniae Testing Therapeutic Intervention Thrombin Transforming Growth Factor beta Transforming Growth Factors Tuberculosis Up-Regulation base calretinin cell motility cell type clinically significant cytokine effusion epithelial to mesenchymal transition glycogen synthase kinase 3 beta in vivo injury and repair irradiation knock-down migration new therapeutic target novel novel therapeutics prevent pulmonary function repaired slug systemic inflammatory response targeted treatment transcription factor

项目摘要

Pleural fibrosis is the scarring of the pleura resulting in restrictive lung disease and impaired lung function. The pathophysiological mechanism of pleural fibrosis is unclear. The interactions between resident and inflammatory cells, profibrotic mediators and coagulation factors, and fibrinolytic pathways are integral to pleural remodeling and fibrosis. Increasing evidence affirm the critical role of pleural mesothelial cells (PMCs) in pleural fibrosis development, mainly through a process termed mesothelial to mesenchymal transition (MesoMT). MesoMT is characterized by increased expression of α-smooth muscle actin (α-SMA)/collagen 1 (Col-1)/fibronectin (FN), and enhanced cell migration/invasion. Currently, there are no pharmacologic treatments for this disease. Therefore, identification of novel targets and therapeutic strategies is an important goal for the public health. However, there is a fundamental knowledge gap in mechanisms controlling MesoMT during pleural fibrosis. Our preliminary data strongly support that dedicator of cytokinesis 2 (DOCK2) is a crucial regulator of MesoMT to promote pleural fibrosis. In primary human PMCs (HPMCs), DOCK2 was induced by the potent MesoMT inducer TGF-β. DOCK2 knockdown blocked TGFβ-induced MesoMT maker expression and cell migration. Snail as a transcriptional factor controlling epithelial to mesenchymal transition was found critical in TGF-β-induced MesoMT. DOCK2 knockdown inhibited TGF-β-induced Snail expression and activation of Smad2/3 and NF-κB signaling, which have been shown to upregulate Snail expression in various cell types. In addition, we found that DOCK2 was dramatically induced in the fibrotic pleura of human pleuritis patients and in pleural fibrosis models induced by Streptococcus pneumoniae (Strep), carbon black/bleomycin (CBB), and TGF-β. DOCK2 knockout mice were significantly protected from Strep-induced pleural fibrosis. Based on these findings, our overall hypothesis is that DOCK2 mediates MesoMT and increases PMC migration/invasion to promote pleural fibrosis, which will be tested in three specific aims. In Aim 1, we will determine if DOCK2 promotes pleural MesoMT via upregulation of Snail. Further, we will test whether DOCK2 increases Snail through activating Smad2/3 and NF-κB signaling. In Aim 2, we will test if DOCK2 promote pleural MesoMT with increased cell migration/invasion. Specifically, we will determine if DOCK2 mediates TGF-β-induced cytoskeletal reorganization, migration/invasion via activating Rac1. In Aim 3, we will test the hypothesis that DOCK2 knockout blocks pleural fibrosis via inhibiting MesoMT in vivo. We will determine if general and mesothelial cell-specific DOCK2 knockout mice are protected from Strep, CBB, and TGF-β induced pleural fibrosis through suppressing MesoMT in vivo. Completion of the proposed studies will establish the pivotal role and mechanisms of DOCK2 in promoting pleural fibrosis by regulating MesoMT, which may ultimately contribute to the identification of novel targeted therapies for this important but refractory clinical problem.

胸膜纤维化是胸膜的疤痕，导致限制性肺病和肺功能受损。胸膜纤维化的病理生理机制尚不清楚。炎症细胞、促纤维化介质和凝血因子以及纤溶途径是不可或缺的越来越多的证据证实了胸膜间皮细胞（PMC）的关键作用。在胸膜纤维化发展过程中，主要通过称为间皮向间质转化的过程 (MesoMT) 的特点是 α-平滑肌肌动蛋白 (α-SMA)/胶原蛋白 1 的表达增加。 (Col-1)/纤连蛋白(FN)，增强细胞迁移/侵袭，目前尚无药理作用。因此，确定新的靶点和治疗策略非常重要。然而，在控制 MesoMT 的机制方面存在基本的知识差距。我们的初步数据强烈支持胞质分裂贡献者 2 (DOCK2) 是一种 DOCK2 是 MesoMT 促进胸膜纤维化的关键调节因子。 DOCK2 敲低可阻断 TGFβ 诱导的 MesoMT 标记物 Snail 作为控制上皮间质转化的转录因子。发现 DOCK2 敲低对 TGF-β 诱导的 MesoMT 抑制 TGF-β 诱导的 Snail 表达至关重要。以及 Smad2/3 和 NF-κB 信号传导的激活，这些信号已被证明可以上调 Snail 的表达此外，我们发现 DOCK2 在人类纤维化胸膜中被显着诱导。胸膜炎患者和由肺炎链球菌（Strep）、碳诱导的胸膜纤维化模型黑色/博来霉素 (CBB) 和 TGF-β 敲除小鼠受到显着保护，免受链球菌诱导的影响。基于这些发现，我们的总体假设是 DOCK2 介导 MesoMT 和增加 PMC 迁移/侵袭以促进胸膜纤维化，这将在三个特定目标中进行测试。目标 1，我们将确定 DOCK2 是否通过上调 Snail 促进胸膜 MesoMT。此外，我们将测试。 DOCK2 是否通过激活 Smad2/3 和 NF-κB 信号传导来增加 Snail 在目标 2 中，我们将测试是否。 DOCK2 通过增加细胞迁移/侵袭来促进胸膜 MesoMT。 DOCK2 通过激活 Rac1 介导 TGF-β 诱导的细胞骨架重组、迁移/侵袭。我们将测试 DOCK2 敲除通过抑制体内 MesoMT 来阻止胸膜纤维化的假设。确定普通和间皮细胞特异性 DOCK2 敲除小鼠是否受到保护，免受链球菌、CBB 和 TGF-β 通过抑制体内 MesoMT 诱导胸膜纤维化完成拟议的研究。建立DOCK2通过调节MesoMT促进胸膜纤维化的关键作用和机制，这可能最终有助于针对这一重要但难治性的疾病确定新的靶向疗法临床问题。