The PINCH-ILK-parvin complexes in glomerular cells

肾小球细胞中的 PINCH-ILK-parvin 复合物

• 批准号: 7903720
• 负责人: CHUANYUE WU
• 金额: $ 9.99万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7903720
• 关键词: Adhesions; Albuminuria; Apoptosis; Binding; Biological Assay; Bioluminescence; Cell-Matrix Junction; Cells; Cellular Structures; Complex; Cytoskeleton; Defect; Deposition; Energy Transfer; Extracellular Matrix; Failure; Fibronectins; Funding; Genes; Goals; Kidney Failure; Knock-out; Knockout Mice; Lead; Life; MAPK8 gene; Mediating; Molecular; Molecular Genetics; Mus; Pathogenesis; Phosphorylation; Process; Proline; Regulation; Role; Shapes; Signal Transduction; Testing; Time; base; cell behavior; human RSU1 protein; human TGFB1 protein; in vivo; integrin-linked kinase; mesangial cell; mitogen-activated protein kinase p38; novel; novel therapeutic intervention; podocyte; protein complex; protein expression

DESCRIPTION (provided by applicant): The long term goal of this competing continuation application is to elucidate the molecular mechanisms by which glomerular cells control extracellular matrix deposition, adhesion, cytoskeleton and apoptosis. Integrin-linked kinase (ILK) is a cytoplasmic component of the cell-matrix adhesions implicated in progressive glomerular failure. Our studies during the previous funding period have demonstrated that ILK forms a complex with PINCH-1 and alpha-parvin. This ternary protein complex appears to function as a key part of the cellular machinery at cell-matrix adhesions that regulates glomerular cell behavior. The proposed studies focus on three aspects of this protein complex in glomerular cells, namely its upstream regulators, downstream effectors, and in vivo functions. Aim 1 is to investigate the mechanism by which the complex is regulated in mesangial cells and podocytes. Our recent studies identify TGF-beta1 as an upstream regulator of the complex but the mechanisms are not known. We will analyze in real time the regulation of this complex in live glomerular cells, and to investigate the mechanisms whereby TGF-beta1 regulates this complex. Aim 2 is to characterize the downstream effectors through which the PINCH-1-ILK-alpha-parvin complex regulates mesangial fibronectin matrix deposition, podocyte-matrix adhesion, cytoskeleton and survival. Recent studies by us and others have identified several candidates. We will use a combination of genetic, molecular and cellular approaches to define their roles in the PINCH-1-ILK-alpha-parvin complex-mediated processes in glomerular cells. Aim 3 is to determine the functions of PINCH-1 in podocytes in vivo. Based on recent studies by us and others, we hypothesize that loss of PINCH-1 in vivo will compromise podocyte cytoskeletal regulation, matrix organization, adhesion, and survival signaling. We will test this hypothesis by selectively inactivating the PINCH-1 gene in mouse podocytes in vivo. Furthermore, we will generate conditionally immortalized PINCH-1flox/flox podocytes, and use them to further investigate the mechanism by which PINCH-1 functions in podocytes. Glomerular damage is a main reason of renal failure. The proposed studies, therefore, not only will advance our understanding of the molecular mechanisms that govern glomerular cell behavior but also may lead to novel therapeutic approaches that alleviate progressive glomerular failure.

描述（由申请人提供）：本竞争性延续申请的长期目标是阐明肾小球细胞控制细胞外基质沉积、粘附、细胞骨架和凋亡的分子机制。整合素连接激酶 (ILK) 是细胞基质粘附的细胞质成分，与进行性肾小球衰竭有关。我们在上一个资助期间的研究表明，ILK 与 PINCH-1 和 alpha-parvin 形成复合物。这种三元蛋白质复合物似乎是调节肾小球细胞行为的细胞基质粘附细胞机制的关键部分。拟议的研究重点关注肾小球细胞中这种蛋白质复合物的三个方面，即其上游调节因子、下游效应因子和体内功能。目标 1 是研究该复合物在系膜细胞和足细胞中的调节机制。我们最近的研究确定 TGF-β1 是该复合物的上游调节因子，但其机制尚不清楚。我们将实时分析活体肾小球细胞中该复合物的调节情况，并研究 TGF-β1 调节该复合物的机制。目标 2 是表征下游效应子，PINCH-1-ILK-α-parvin 复合物通过这些效应子调节系膜纤连蛋白基质沉积、足细胞-基质粘附、细胞骨架和存活。我们和其他人最近的研究已经确定了几个候选者。我们将结合遗传、分子和细胞方法来确定它们在肾小球细胞中 PINCH-1-ILK-α-parvin 复合物介导的过程中的作用。目的 3 是确定 PINCH-1 在体内足细胞中的功能。根据我们和其他人最近的研究，我们假设体内 PINCH-1 的缺失将损害足细胞的细胞骨架调节、基质组织、粘附和生存信号传导。我们将通过选择性地灭活体内小鼠足细胞中的 PINCH-1 基因来检验这一假设。此外，我们将生成条件永生化的PINCH-1flox/flox足细胞，并利用它们进一步研究PINCH-1在足细胞中发挥作用的机制。肾小球损伤是肾功能衰竭的主要原因。因此，拟议的研究不仅将增进我们对控制肾小球细胞行为的分子机制的理解，而且可能会带来减轻进行性肾小球衰竭的新治疗方法。