Kindlin-2 in Cell-Matrix Adhesion and Signaling

Kindlin-2 在细胞基质粘附和信号传导中的作用

基本信息

批准号：
8040189
负责人：
CHUANYUE WU
金额：
$ 32.67万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-05-01 至 2014-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8040189
关键词：
Abnormal Cell Adhesions Behavior Binding Biology Catalytic Domain Cell-Matrix Junction Cytoplasmic Protein Cytoskeleton Defect Dominant-Negative Mutation Event Extracellular Matrix Focal Adhesions Funding Genetic Goals Growth Hepatocyte Integrins Knock-in Mouse Knock-out Knockout Mice Lead Link Liver Malignant Neoplasms Mediating Membrane Membrane Lipids Membrane Proteins Molecular Natural regeneration Organ PTEN gene Pathogenesis Pathologic Processes Phosphatidylinositols Phosphotransferases Physiological Physiological Processes Process Protein Binding Proteins Regulation Role Scaffolding Protein Signal Transduction Site Structure Testing Tissues Work base cell behavior design disorder control human disease improved injury and repair integrin-linked kinase molecular pathology mutant new therapeutic target novel strategies phosphatidylinositol 3,4,5-triphosphate phosphoinositide-3,4,5-triphosphate receptor tissue processing tissue regeneration

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this competing renewal application is to elucidate the molecular basis underlying cell- extracellular matrix (ECM) adhesion and regulation, and the mechanism whereby they control cell behavior, tissue integrity, growth and regeneration. Recent studies by the applicant and others have demonstrated a critical role of kindlin-2 (also known as Mig-2), a widely expressed membrane-cytoskeleton junctional protein, in integrin activation and cell-ECM adhesion. How kindlin-2 regulates these processes, however, is not known. Based on findings obtained during previous project periods, the applicant hypothesizes that kindlin-2 regulates these processes through interacting with membrane lipids and protein components of cell-ECM adhesions. To test this hypothesis, he proposes studies with the following three aims. Aim 1 is to characterize the interaction of kindlin-2 with membrane lipids and assess its role in regulation of integrins and integrin-dependent processes. To this end, he will employ genetic, pharmacological and dominant negative inhibition strategies to ablate this interaction, and determine the consequences. Aim 2 is to determine the functions of kindlin-2 interactions with focal adhesion proteins in regulation of cell-ECM adhesion. He will define the sites mediating the interactions and use a "knock-in" strategy to replace wild type kindlin-2 with mutants lacking specific protein-binding activity and determine the consequences. Aim 3 is to investigate the functions of kindlin-2 and its interplay with ILK in liver structure, growth and regeneration, which are known to be regulated by ECM adhesion and ILK signaling. He will generate hepatocyte-specific kindlin-2 knockout and "knock-in" mice, in which wild type kindlin-2 is substituted with kindlin-2 mutants lacking specific binding activities, and determine contributions of kindlin-2 and its interactions to regulation of hepatocyte behavior, liver structure, growth and regeneration. These studies will fill important gaps in our understanding of the mechanism whereby cell-ECM adhesion and ECM-dependent tissue processes are regulated. Given the importance of cell-ECM adhesion in human diseases, these studies may also lead to novel approaches to control diseases associated with abnormal cell-ECM adhesion and signaling. PUBLIC HEALTH RELEVANCE: Alteration of cell-ECM adhesion is critically involved in the pathogenesis of human diseases including cancer. This project seeks to determine how a recently identified regulator of cell-ECM adhesion influences cell-ECM adhesion, tissue structure, growth and regeneration. These studies may lead to identification of novel therapeutic targets to control diseases associated with abnormal cell-ECM adhesion and growth.

描述（由申请人提供）：本次竞争性更新申请的长期目标是阐明细胞-细胞外基质（ECM）粘附和调节的分子基础，以及它们控制细胞行为、组织完整性、生长和再生的机制。申请人和其他人最近的研究证明了kindlin-2（也称为Mig-2）（一种广泛表达的膜-细胞骨架连接蛋白）在整合素激活和细胞-ECM粘附中的关键作用。然而，kindlin-2 如何调节这些过程尚不清楚。根据之前项目期间获得的发现，申请人假设 kindlin-2 通过与细胞 ECM 粘附的膜脂和蛋白质成分相互作用来调节这些过程。为了检验这一假设，他提出了以下三个目标的研究。目标 1 是表征 kindlin-2 与膜脂的相互作用，并评估其在调节整合素和整合素依赖性过程中的作用。为此，他将采用遗传、药理学和显性负抑制策略来消除这种相互作用，并确定后果。目标 2 是确定 kindlin-2 与粘着斑蛋白相互作用在调节细胞-ECM 粘附中的功能。他将定义介导相互作用的位点，并使用“敲入”策略将野生型 kindlin-2 替换为缺乏特定蛋白质结合活性的突变体，并确定后果。目标 3 是研究 kindlin-2 的功能及其与 ILK 在肝脏结构、生长和再生中的相互作用，已知这些功能受到 ECM 粘附和 ILK 信号传导的调节。他将培育肝细胞特异性 kindlin-2 敲除和“敲入”小鼠，其中野生型 kindlin-2 被缺乏特异性结合活性的 kindlin-2 突变体取代，并确定 kindlin-2 的贡献及其相互作用对调节肝细胞行为、肝脏结构、生长和再生。这些研究将填补我们对细胞-ECM 粘附和 ECM 依赖性组织过程调节机制的理解的重要空白。鉴于细胞-ECM 粘附在人类疾病中的重要性，这些研究还可能带来控制与异常细胞-ECM 粘附和信号传导相关疾病的新方法。公共卫生相关性：细胞-ECM 粘附的改变与包括癌症在内的人类疾病的发病机制密切相关。该项目旨在确定最近发现的细胞-ECM 粘附调节剂如何影响细胞-ECM 粘附、组织结构、生长和再生。这些研究可能会导致确定新的治疗靶点，以控制与异常细胞-ECM 粘附和生长相关的疾病。