Elucidating tensin1-associated signals that promote fibronectin matrix assembly and lung fibrosis

项目摘要： 特发性肺纤维化(IPF)的特征是肺部不断形成疤痕和僵硬，导致 确诊后3-4年内死亡。随着IPF在美国每年导致4万人死亡，新的治疗方法 是阻止这种疾病突然发展的迫切需要。我们已经发现，焦点黏附基因， 紧张素(TNS1)在肌成纤维细胞分化、形成新的细胞外基质(ECM)等过程中发挥作用。 肺纤维化的进展。然而，目前尚不清楚TNS1如何介导基质重塑和肺组织。 功能衰退。我们的理论基础是确定TNS1在肌成纤维细胞分化和分化中的作用。 肺纤维化的发展将使我们能够直接靶向新的ECM形成，从而阻断进展。 这种疾病的危害。我们假设TNS1与粘着斑激酶(FAK)和Rho依赖的蛋白有协同作用 体内诱导肌成纤维细胞分化和肺纤维化的信号。为了验证这一假设，我们将： 目的1.在体内确定TNS1如何改变损伤的修复反应。人类的主要发现 疾病和体外模型提示TNS1在基质组装和肺肿瘤进展中起关键作用 纤维化症。在这个目标中，我们将确定TNS1如何修改体内对损伤的修复反应。要做到这一点， 我们将在损伤/修复的活体模型(博莱霉素模型)中利用TNS1的条件缺失来确定 TNS1相关信号如何改变早期损伤反应、全局修复反应和细胞命运 维修过程中的决心。 目的2.确定TNS1用于促进纤维连接蛋白基质组装的协同信号和 肌成纤维细胞分化。我们假设TNS1可能与Rho和FAK合作改变细胞命运， 和整合素连接的激酶信号，以促进纤维连接蛋白(FN)基质组装。为了阐明这些途径 参与FN基质组装，我们将利用人肺原代细胞培养和小鼠肺细胞培养。 TnS1f，f小鼠，以确定TNS1如何定位于黏附复合体，修改Rho，ILK和FAK信号， 并促进纤维粘连的形成、FN纤维的组装和ECM的沉积。 在这些研究完成后，我们将确定修改修复性的关键信号事件 对损伤的反应和改变FN基质组装的关键信号通路。这一知识将提供 干扰细胞介导的基质形成的干预靶点，从而扩大我们的治疗选择 体内抗肺纤维化实验。作为一名内科科学家，他既致力于治疗患有 肺纤维化，并对肌成纤维细胞的调节机制进行了卓有成效的研究 发展、基质组装和体内纤维化，我准备好领导这项研究，并将这项工作带到 完成了。

Project Abstract: Idiopathic pulmonary fibrosis (IPF) is characterized by unrelenting scarring and stiffening of the lungs that leads to death within 3-4 years after diagnosis. As IPF kills 40,000 individuals in the U.S. each year, new treatments are urgently needed to abrupt the progression of this disease. We have found that the focal adhesion gene, tensin (TNS1), plays a role in myofibroblast differentiation, the formation of new extracellular matrix (ECM), and the progression of pulmonary fibrosis. However, it is not known how TNS1 mediates matrix remodeling and lung function decline. Our rationale is that determining the role of TNS1 in myofibroblast differentiation and development of pulmonary fibrosis will allow us to directly target new ECM formation, thus disrupting progression of the disease. We hypothesize that TNS1 cooperates with focal adhesion kinase (FAK) and Rho-dependent signals to induce myofibroblast differentiation and pulmonary fibrosis in vivo. To test this hypothesis, we will: Aim 1. Determine how TNS1 modifies the reparative response to injury in vivo. Key findings in human disease and in vitro models suggest a key role for TNS1 in matrix assembly and the progression of pulmonary fibrosis. In this aim, we will determine how TNS1 modifies the in vivo reparative response to injury. To do so, we will utilize conditional deletion of TNS1 in an in vivo model of injury/repair (bleomycin model) to determine how TNS1-associated signaling modifies early injury responses, global reparative response, and cell fate determination during repair. Aim 2. Determine cooperative signaling used by TNS1 to promote fibronectin matrix assembly and myofibroblast differentiation. We hypothesize that TNS1 may cooperate with Rho and FAK to modify cell fate, and integrin-linked kinase signaling to facilitate fibronectin (FN) matrix assembly. To elucidate the pathways involved in FN matrix assembly, we will utilize human lung primary cell cultures and murine cell cultures from TNS1f,f mice to determine how TNS1 localizes to adhesive complexes, modifies Rho, ILK, and FAK signaling, and facilitates fibrillar adhesion formation, FN fibril assembly, and ECM deposition. Upon completion of these studies, we will have determined key signaling events in modifying the reparative response to injury and key signaling pathways that modify FN matrix assembly. This knowledge will provide targets for intervention to disrupt cell-mediated matrix formation, thereby expanding our therapeutic options to halt pulmonary fibrosis in vivo. As a physician-scientist who is both dedicated to the care of patients with pulmonary fibrosis and has a track-record of productive investigation of mechanisms mediating myofibroblast development, matrix assembly, and in vivo fibrosis, I am well-poised to lead this study and bring this work to completion.