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

• 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
• 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

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）：该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。申请人和其他人的最近研究已经证明了kindlin-2（也称为Mig-2）（一种广泛表达的膜-细胞骨架连接蛋白）在整联蛋白活化和细胞-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。基于在先前项目期间获得的发现，申请人假设kindlin-2通过与细胞-ECM粘附的膜脂质和蛋白质组分相互作用来调节这些过程。为了验证这一假设，他提出了以下三个目标的研究。目的1是表征kindlin-2与膜脂质的相互作用，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、药理和显性负抑制策略来消除这种相互作用，并确定其后果。目的二是研究kindlin-2与黏着斑蛋白的相互作用对细胞与ECM粘附的调节作用。他将确定介导相互作用的位点，并使用“敲入”策略用缺乏特异性蛋白结合活性的突变体取代野生型kindlin-2，并确定结果。目的3：研究Kindlin-2在肝脏结构、生长和再生中的作用及其与ILK的相互作用。他将产生肝细胞特异性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及其相互作用对肝细胞行为，肝脏结构，生长和再生的调节的贡献。这些研究将填补我们对细胞-细胞外基质粘附和细胞外基质依赖性组织过程调节机制理解的重要空白。鉴于细胞-ECM粘附在人类疾病中的重要性，这些研究也可能导致控制与异常细胞-ECM粘附和信号传导相关的疾病的新方法。 公共卫生相关性：细胞-ECM粘附的改变与人类疾病（包括癌症）的发病机制密切相关。该项目旨在确定最近确定的细胞-ECM粘附调节剂如何影响细胞-ECM粘附，组织结构，生长和再生。这些研究可能导致识别新的治疗靶点，以控制与异常细胞-ECM粘附和生长相关的疾病。