PINCH-1 Interactions and Functions

PINCH-1 相互作用和功能

基本信息

批准号：
7534806
负责人：
CHUANYUE WU
金额：
$ 28.09万
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 2010-11-30
项目状态：
已结题

项目摘要

Cell-extracellular matrix (ECM) adhesion and signaling are essential for tissue integrity and organ functions. They regulate a variety of processes including cytoskeletal organization, cell migration and survival. Alterations of cell-ECM adhesion and signaling are intimately associated with human diseases. The long-term goal of this competing continuation application is to determine the molecular basis underlying cell- ECM adhesion and signaling, and the mechanism whereby they control cell behavior and organ function. PINCH-1 is a widely expressed and evolutionally conserved component of cell-ECM adhesions. Recent studies have revealed important roles of PINCH-1 in regulation of actin cytoskeleton, cell migration and apoptosis. Despite recent progress, our understanding of the molecular mechanisms by which PINCH-1 functions is still quite primitive. Furthermore, the roles of PINCH-1 in organ biology remain largely unknown. To fill these gaps, we propose studies with the following specific aims. Aim 1 is to investigate the mechanisms by which PINCH-1 regulates actin cytoskeleton and cell migration. Our hypothesis is that PINCH-1 plays both a structural and a signaling role in its regulation of actin cytoskeleton and cell migration. Aim 2 is to investigate the mechanism by which PINCH-1 regulates apoptosis. We will identify the binding partners and downstream signaling intermediates through which PINCH-1 regulates this process. Aim 3 is to determine the functions of PINCH-1 in liver biology. We will generate liver PINCH-1 knockout mice using the Cre-lox system. The consequences of ablation of PINCH-1 expression in the liver will be determined by molecular, cellular, histological and functional analyses. Furthermore, we will compare the phenotypes with those induced by elimination of ILK in the liver. Finally, we will prepare primary hepatocytes and test whether PINCH-1 protects them from anoikis, a major obstacle for hepatocyte transplantation. The proposed studies will shed light on the organ biology and molecular mechanism of PINCH-1, a key component of cell-ECM adhesions. Furthermore, they will help us to better understand the general mechanism governing cell-ECM adhesion-mediated cytoskeletal regulation, cell migration and apoptosis. Ultimately, these studies may lead to novel therapeutic approaches to control pathological processes in the liver and other organs that are associated with abnormal cell-ECM adhesion, migration and apoptosis.

细胞-细胞外基质 (ECM) 粘附和信号传导对于组织完整性和器官至关重要功能。它们调节多种过程，包括细胞骨架组织、细胞迁移和生存。细胞-ECM 粘附和信号传导的改变与人类疾病密切相关。这这一竞争性延续应用的长期目标是确定细胞的分子基础 ECM 粘附和信号传导，以及它们控制细胞行为和器官功能的机制。 PINCH-1 是细胞-ECM 粘附中广泛表达且进化保守的成分。最近的研究揭示了 PINCH-1 在调节肌动蛋白细胞骨架、细胞迁移和细胞凋亡。尽管最近取得了进展，但我们对 PINCH-1 分子机制的理解功能还是很原始。此外，PINCH-1 在器官生物学中的作用仍然很大程度上未知。到为了填补这些空白，我们提出了以下具体目标的研究。目标 1 是通过以下方式研究其机制：其中 PINCH-1 调节肌动蛋白细胞骨架和细胞迁移。我们的假设是 PINCH-1 既扮演肌动蛋白细胞骨架和细胞迁移的调节中的结构和信号传导作用。目标2是调查 PINCH-1 调节细胞凋亡的机制。我们将确定绑定合作伙伴和下游 PINCH-1 通过其调节该过程的信号传导中间体。目标 3 是确定以下功能肝脏生物学中的 PINCH-1。我们将使用 Cre-lox 系统生成肝脏 PINCH-1 基因敲除小鼠。这肝脏中 PINCH-1 表达消除的后果将由分子、细胞、组织学和功能分析。此外，我们将比较表型与诱导的表型消除肝脏中的ILK。最后，我们将制备原代肝细胞并测试PINCH-1是否具有保护作用。它们来自失巢凋亡，失巢凋亡是肝细胞移植的主要障碍。拟议的研究将阐明 PINCH-1 的器官生物学和分子机制，PINCH-1 是一种细胞-ECM 粘附的关键组成部分。此外，它们将帮助我们更好地了解一般情况。控制细胞-ECM粘附介导的细胞骨架调节、细胞迁移和凋亡的机制。最终，这些研究可能会带来控制病理过程的新治疗方法。肝脏和其他与异常细胞 ECM 粘附、迁移和凋亡相关的器官。