Cell Adhesion Mechanisms in Vascular Disease & Thrombosis

需要支持继续一项旨在促进对血管疾病分子机制的理解的计划，并通过一群经验丰富的科学家的合作努力，促进新的诊断、治疗和预防策略的开发，这些科学家专注于细胞黏附的统一主题。这个跨学科的项目将跨越生物化学、细胞生物学、体外和体内研究，以评估血流对黏附和信号的影响，以及对转基因小鼠和斑马鱼的分析。在项目1中，金斯伯格博士将研究整合素的激活以及由此导致的血小板血栓形成和白细胞的抑制。具体的研究将涉及纯化系统中整合素的激活，RIAM的作用，整合素跨膜区的相互作用和结构，以及干扰血小板和白细胞整合素激活的体内和体外后果。在项目2中，Shattil博士将继续开发和利用各种策略来可视化活细胞中蛋白质与血小板整合素αLibβ3的相互作用，并将在斑马鱼体内分析αV整合素的信号机制。在项目3中，Ruggeri博士将在von Willebrand因子(VWF)和血小板GPIB结构方面的进展的基础上再接再厉。具体地说，他将寻求更好地了解流动血液中GPIB和VWF之间的键的生物力学特性，凝血酶对GPlb/VWF相互作用的影响，以及一种新发现的免疫球蛋白调节剂在血栓形成中的生物学作用。由Petrich博士领导的小鼠遗传学核心单元A将提供专业知识、基因组结构、基因分型、具有良好特性的小鼠胚胎干细胞，以及用于小鼠遗传操作目的的囊胚注射。这个核心将被用来产生有选择性地扰乱整合素激活的Talin敲打基因，并将提供有条件的转基因动物。由物理学家格罗伊斯曼博士领导的微流控核心B单元将开发和提供高通量多通道微流控流动系统，以分析在受控剪应力下血小板和白细胞与传统和有图案的基质的粘附性。行政核心股C将继续提供行政支助。总而言之，这一跨学科计划将使一群经验丰富的研究人员之间实现显著的协同作用，他们将测试新的假设，并利用和开发尖端方法来促进对血管生物学和血栓形成中细胞黏附事件的了解。

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.