ILK Signaling in Renal Growth and Matrix Deposition

描述(申请人提供)：肾(例如，肾小球系膜)细胞黏附、生长和细胞外基质沉积在终末期肾功能衰竭的进展中起关键作用。这种竞争性持续应用的长期目标是阐明肾细胞控制这些过程的分子机制。在这项竞争性的延续申请中提出的研究集中在整合素连接激酶(ILK)在调节肾小球系膜细胞行为中的作用，ILK是细胞-基质接触部位的关键成分。目的1验证以下假设：PINCH/ILKICH-ILKBP复合体在整合素和肾小球系膜细胞-基质接触部位的肌动蛋白细胞骨架之间提供关键连接，从而调节系膜细胞的黏附、生长和纤维连接蛋白基质沉积。通过腺病毒感染将干扰PINCH/ILKICH-ILKBP复合体形成的显性负性抑制物引入原代系膜细胞，并将确定它们对系膜细胞黏附、增殖和纤维连接蛋白基质沉积的影响。目的2验证PINCH-RP是一种新发现的PINCH相关蛋白，其在肾小球系膜细胞中的表达受葡萄糖的调节，是系膜细胞中P1INCH/ILKICH-ILKBP复合体组装和功能的天然负调节因子。分子和细胞方法的结合将被用来确定P1NCH-RP在系膜细胞中的功能。目的3是对Mig-2的分子特征进行鉴定，Mig-2是一种新发现的蛋白，它与Pinch/ILKICH-ILKBP复合体共定位并物理结合，并确定它是否具有促进ILK复合体在系膜细胞中定位和功能的正调控功能。这些研究将促进我们对控制肾小球系膜细胞黏附、生长和细胞外基质沉积的分子机制的了解。它们可能导致开发新的治疗方法来控制与异常肾细胞黏附、增殖和细胞外基质沉积密切相关的终末期肾功能衰竭。

DESCRIPTION (provided by applicant): Renal (e.g., glomerular mesangial) cell adhesion, growth and extracellular matrix deposition are crucially involved in the progression of end-stage renal failure. The long-term goal of this competing continuation application is to elucidate the molecular mechanism by which renal cells control these processes. The studies proposed in this competing continuation application focus on the role of integrin-linked kinase (ILK), a key component of cell-matrix contact sites, in the regulation of glomerular mesangial cell behavior. Aim 1 is to test the hypothesis that a PINCH/ILKICH-ILKBP complex provides a key connection between integrins and the actin cytoskeleton at glomerular mesangial cell-matrix contact sites and thereby regulates mesangial cell adhesion, growth and fibronectin matrix deposition. Dominant negative inhibitors that disrupt the formation of the PINCH/ILKICH-ILKBP complex will be introduced into primary mesangial cells by adenoviral infection and their effects on mesangial cell adhesion, proliferation and fibronectin matrix deposition will be determined. Aim 2 is to test the hypothesis that PINCH-RP, a newly identified PiNCH-related protein whose expression in glomerular mesangial cells is regulated by glucose, is a naturally occurring negative regulator of the assembly and functions of the P1INCH/ILKICH-ILKBP complex in mesangial cells. A combination of molecular and cellular approaches will be used to determine the functions of P1NCH-RP in mesangial cells. Aim 3 is to molecularly characterize Mig-2, a newly identified protein that co-localizes and physically associates with the PINCH/ILKICH-ILKBP complex, and to determine whether it functions as a positive regulator facilitating the localization and functions of the ILK complex in mesangial cells. These studies will advance our knowledge on the molecular mechanisms that govern glomerular mesangial cell adhesion, growth and extracellular matrix deposition. They could potentially lead to development of novel therapeutic approaches to control end-stage renal failure that is intimately associated with abnormal renal cell adhesion, proliferation and extracellular matrix deposition.