Cell Adhesion Mechanisms in Vascular Disease & Thrombosis

本项目旨在促进对血管疾病分子机制的理解，并通过一组专注于细胞粘附统一主题的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。这个跨学科的项目将跨越生物化学、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阻滞导致的血栓形成。具体的研究将涉及整合素在纯化系统中的激活，RIAM的作用，整合素跨膜结构域的相互作用和结构，以及扰乱整合素在血小板和白细胞中的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策略来可视化蛋白质与活细胞中血小板整合素α lIb β 3的相互作用，他将分析斑马鱼体内α V整合素的信号传导机制。在项目3中，Ruggeri博士将以血管性血液病因子（VWF）和血小板GPIb结构的进展为基础。具体来说，他将寻求更好地理解流动血液中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，以及新发现的免疫球蛋白对血栓形成的生物学作用。Mouse Genetics Core Unit A，由Dr. Petrich领导，将提供专业知识，基因组构建，基因分型，具有良好特征的小鼠胚胎干细胞，以及用于小鼠遗传操作的囊胚注射。该核心将用于产生talin敲除蛋白，选择性地干扰整合素的激活，并将提供有条件的转基因动物。由物理学家Groisman博士领导的微流体核心单元B将开发并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对传统和图案基板的粘附。行政核心股C将继续提供行政支助。总的来说，这个跨学科的项目将使一群有成就的研究者之间产生显著的协同作用，他们将测试新的假设，利用和开发尖端的方法来推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.