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，导致严格的肺部疾病和肺功能受损。胸膜纤维化的病理生理机制尚不清楚。居民与炎性细胞，纤维化介质和凝结因子以及纤维蛋白水解途径是不可或缺的胸膜重塑和纤维化。越来越多的证据肯定了胸膜间皮细胞（PMC）的关键作用在胸膜纤维化发育中，主要通过称为间质转变的过程（Mesomt）。 Mesomt的特征是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），并增强细胞迁移/侵袭。目前，没有药理学治疗这种疾病。因此，识别新目标和治疗策略是重要的公共卫生的目标。但是，控制Mesomt的机制存在基本知识差距在胸膜纤维化期间。我们的初步数据强烈支持Cytokinesis 2（Dock2）的奉献者是一个中膜的关键调节剂，以促进胸膜纤维化。在主要人类PMC（HPMC）中，Dock2是由潜在的介子诱导的TGF-β诱导。 DOCK2敲低阻断TGFβ诱导的Mesomt Maker 表达和细胞迁移。蜗牛作为控制上皮过渡上皮的转录因子在TGF-β诱导的中瘤中发现至关重要。 DOCK2敲低抑制了TGF-β诱导的蜗牛表达 SMAD2/3和NF-κB信号的激活，已证明可以更新蜗牛表达各种细胞类型。此外，我们发现DOCK2在人类的纤维性胸膜中急剧诱导胸膜炎患者和肺炎链球菌诱导的胸膜纤维化模型（链球菌），碳黑色/博来霉素（CBB）和TGF-β。 Dock2敲除小鼠受到链球菌诱导的胸膜纤维化。基于这些发现，我们的总体假设是Dock2介导了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。 AIM 1，我们将确定Dock2是否通过上调蜗牛来促进胸膜膜。此外，我们将测试 DOCK2是否通过激活SMAD2/3和NF-κB信号传导增加蜗牛。在AIM 2中，我们将测试是否 DOCK2随着细胞迁移/侵袭的增加而促进胸膜中膜。具体来说，我们将确定是否 DOCK2介导TGF-β诱导的细胞骨架重组，通过激活Rac1进行迁移/侵袭。在AIM 3中，我们将检验以下假设：Dock2敲除通过抑制体内膜的胸膜纤维化阻断胸膜纤维化。我们将确定一般和间皮细胞特异性的Dock2基因敲除小鼠是否受到链球菌，CBB和 TGF-β通过在体内抑制Mesomt诱导胸膜纤维化。拟议研究的完成将通过调节Mesomt，建立DOCK2在促进胸膜纤维化中的关键作用和机制最终可能有助于确定这种重要但难治性的新型靶向疗法临床问题。