PINCH-1 Interactions and Functions

描述（由申请人提供）：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功能至关重要。它们调节多种过程，包括细胞骨架组织、细胞迁移和存活。细胞-ECM粘附和信号传导的改变与人类疾病密切相关。这种竞争性延续应用的长期目标是确定细胞- ECM粘附和信号传导的分子基础，以及它们控制细胞行为和器官功能的机制。PINCH-1是细胞-ECM粘附的广泛表达和进化保守的组分。最近的研究表明PINCH-1在调节肌动蛋白细胞骨架、细胞迁移和凋亡中发挥重要作用。尽管最近取得了进展，但我们对PINCH-1功能的分子机制的理解仍然非常原始。此外，PINCH-1在器官生物学中的作用在很大程度上仍然未知。为了填补这些空白，我们提出了具有以下具体目标的研究。目的1探讨PINCH-1对肌动蛋白骨架和细胞迁移的调控机制。我们的假设是，PINCH-1在其调节肌动蛋白细胞骨架和细胞迁移中起着结构和信号作用。目的二是探讨PINCH-1调控细胞凋亡的机制。我们将确定PINCH-1调节这一过程的结合伙伴和下游信号中间体。目的3：研究PINCH-1在肝脏生物学中的作用。我们将使用Cre-lox系统产生肝脏PINCH-1敲除小鼠。将通过分子、细胞、组织学和功能分析确定肝脏中PINCH-1表达消融的结果。此外，我们将比较表型与肝脏中ILK消除诱导的表型。最后，我们将制备原代肝细胞并测试PINCH-1是否保护它们免受失巢凋亡，这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是细胞-ECM粘附的关键组分。此外，它们将帮助我们更好地理解细胞-ECM粘附介导的细胞骨架调节、细胞迁移和凋亡的一般机制。最终，这些研究可能会导致新的治疗方法，以控制肝脏和其他器官中与异常细胞-ECM粘附，迁移和凋亡相关的病理过程。

DESCRIPTION (provided by applicant): 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.