Cell Adhesion Mechanisms in Vascular Disease & Thrombosis

要求继续支持一项旨在促进对血管疾病分子机制的理解的计划，并通过一组经验丰富的科学家的合作努力，促进新的诊断，治疗和预防策略的发展，重点是细胞粘附的统一主题。这个跨学科的计划将跨越生物化学，细胞生物学，离体和体内研究的学科，以评估血液流动对粘附和信号传导的影响，以及对转基因小鼠和斑马鱼的分析。在项目1中，Ginsberg博士将研究整合素的激活以及血小板和白细胞停滞导致的血栓形成。具体研究将解决纯化系统中整联蛋白的激活、RIAM的作用、整联蛋白跨膜结构域的相互作用和结构，以及扰乱血小板和白细胞中整联蛋白激活的体内和离体后果。在项目2中，Shattil博士将继续开发和利用策略来可视化活细胞中蛋白质与血小板整联蛋白α IIb β 3的相互作用，他将分析斑马鱼体内α V整联蛋白的信号传导机制。在项目3中，Ruggeri博士将建立在血管性血友病因子（VWF）和血小板GPIb结构的基础上。具体而言，他将寻求更好地了解流动血液中GPIb和VWF之间键合的生物力学特性，凝血酶对GPIb/VWF相互作用的影响，以及新发现的血栓形成免疫球蛋白调节剂的生物学作用。由Petrich博士领导的小鼠遗传学核心单位A将提供专门知识、基因组构建、基因分型、充分表征的小鼠胚胎干细胞和囊胚注射，用于小鼠遗传操作。该核心将用于产生选择性干扰整联蛋白活化的talin敲入，并将提供条件性转基因动物。由物理学家Groisman博士领导的微流体核心单元B将开发和提供高通量多通道微流体流动系统，以分析血小板和白细胞在受控剪切应力下与常规和图案化基底的粘附。行政核心股C将继续提供行政支助。总而言之，这个跨学科的计划将使一组有成就的研究人员之间产生显着的协同作用，他们将测试新的假设，并利用和开发尖端的方法，以促进对血管生物学和血栓形成中细胞粘附事件的理解。

Support is requested to continue a program designed to advance understanding of molecular mechanisms of vascular disease and to promote development of new diagnostic, therapeutic, and preventive strategies through the collaborative efforts of a group of experienced scientists focused oh the unifying theme of cell adhesion. This interdisciplinary program will span disciplines of biochemistry, cell biology, ex-vivo and in vivo studies to assess the effects of blood flow on adhesion and signaling, and analysis of genetically-modified mice and zebrafish. In Project 1, Dr. Ginsberg will study the activation of integrins and resulting thrombus formation by platelets and arrest of leukocytes. Specific studies will address the activation of integrins in purified systems, the role of RIAM, the interactions and structure of integrin transmembrane domains, and the in vivo and ex vivo consequences of perturbing integrin activation in platelets and leukocytes. In Project 2, Dr.Shattil will continue to develop and utilize strategies to visualize interactions of proteins with platelet integrin alpha lIb beta 3 in living cells and he will analyze the signaling mechanisms of alpha V integrins in vivo in zebrafish. In Project 3, Dr. Ruggeri will build on advances in the structure of von Willebrand Factor (VWF) and platelet GPIb. Specifically, he will seek to better understand the biomechanical properties of the bonds between GPIb and VWF in flowing blood, the effects of thrombin on the GPlb/VWF interaction, and the biological role of a newly-discovered immunoglobulin modulator of thrombosis. Mouse Genetics Core Unit A, led by Dr. Petrich, will provide expertise, genomic constructs, genotyping, well characterized murine embryonic stem cells, and blastocyst injections for the purpose of genetic manipulation of mice. This core will be used to generate talin knock-ins that selectively perturb integrin activation and will provide conditional transgenic animals. Microfluidics Core Unit B, led by Dr. Groisman, a physicist, will develop and provide high throughput multichannel microfluidic flow systems to analyze platelet and leukocyte adhesion to conventional and patterned substrates under controlled shear stress. Administrative Core Unit C will continue to provide administrative support. Altogether, this interdisciplinary program will enable remarkable synergies amongst a group of accomplished investigators who will test new hypotheses and utilize and develop cutting edge methodologies to advance understanding of cell adhesion events in vascular biology and thrombosis.