Regulation of Vascular Morphogenesis in Adult Brain

成人大脑血管形态发生的调节

• 批准号: 7864039
• 负责人: DONALD R SENGER
• 金额: $ 20.63万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7864039
• 关键词: Address; Adult; Agonist; Architecture; Basement membrane; Blood Vessels; Blood capillaries; Blood flow; Brain; Brain Ischemia; Brain Neoplasms; Calpain; Calpain I; Cells; Clinical; Craniocerebral Trauma; Defect; Development; Disease; Embryo; Embryonic Development; Endothelial Cells; Endothelium; Family; Goals; Guanosine Triphosphate Phosphohydrolases; Health; Hypoxia; Impairment; In Vitro; Injury; Intercellular Junctions; Intervention; Ischemia; Laboratories; Methods; Microscopic; Modeling; Molecular; Morphogenesis; Nervous system structure; Neurologic; Neurological outcome; Neurosurgeon; Outcome; Pathologic Neovascularization; Pathway interactions; Patients; Peptide Hydrolases; Perfusion; Pericytes; Permeability; Pharmaceutical Preparations; Process; Protein Isoforms; Rattus; Recovery; Regulation; Research; Research Personnel; Retina; Signal Pathway; Signal Transduction; Skin; Stimulus; Stroke; Structure; Testing; Trauma; Vascular Diseases; Vascular Endothelial Growth Factors; Work; angiogenesis; base; capillary; cerebrovascular; cytokine; design; experience; improved; in vitro Assay; in vivo; inhibitor/antagonist; mouse model; mutant; neovascularization; nervous system disorder; public health relevance; response; retroviral transduction; rho; rho GTP-Binding Proteins; tool

DESCRIPTION (provided by applicant): The broad long-term goals of the proposed research are to identify basic mechanisms that control vascular morphogenesis in brain and thereby identify new rational drug-based strategies for normalizing pathological blood vessels that form in response to ischemia and trauma. A parallel goal is to understand the molecular and cellular mechanisms responsible for pathological angiogenesis in brain. The working hypothesis for the proposed research is that pathological angiogenesis is a consequence of aberrant vascular morphogenesis and that significant improvement can be achieved with pharmacological intervention, particularly through appropriate targeting of cytoskeletal signaling pathways in the endothelium. The serious defects associated with pathological angiogenesis, including impaired blood flow and vessel hyper-permeability, likely contribute to neurological deficit following ischemic injury. Timely rectification of vascular defects through pharmacological intervention offers the promise of improved neurological outcomes in patients with brain ischemia and trauma. To address these goals, two aims are proposed. The first aim is: In vitro, define signaling pathways that regulate assembly of brain microvascular endothelial cells (BMVECs) into capillary-like structures; and identify a panel of pharmacologically active drugs that modulate these signaling pathways and thereby facilitate vascular morphogenesis. Key goals in this first aim are to define signaling pathways and effector molecules that regulate formation of capillary structures by BMVECs in vitro, with emphasis on Rho GTPases, GSK-3b, and calpain; and to identify drugs that selectively target these pathways and effector molecules in BMVECs and thereby improve formation of capillary structures in vitro. Experimental tools will include retroviral transduction of BMVECs with specific signaling mutants, in vitro capillary morphogenesis assays, in vitro analyses of cell:cell junctions, and pharmacological agents that modulate the activities of Rho GTPases, GSK- 3b, and calpain. The second aim is: In vivo, with a mouse model of VEGF-driven angiogenesis in adult brain, identify rational drug-based strategies for improving both the architecture and function of pathological neovessels. Key goals in this second aim are to identify, with the benefit of findings made in Aim 1, drug-based strategies that normalize aberrant vessel architecture and vascular perfusion, normalize basement membrane and pericyte coverage, and reduce vascular hyper-permeability in brain. Experimental methods will involve a mouse model of pathological angiogenesis in brain, together with microscopic analyses of vascular structure, perfusion, and hyper-permeability. PUBLIC HEALTH RELEVANCE: The goals of this research are to identify basic mechanisms that regulate formation of new blood vessels in adult brain and thereby design new strategies for correcting serious defects commonly associated with disordered blood vessel development. Understanding mechanisms responsible for defective blood vessels in brain is important because stroke and traumatic injury result in formation of blood vessels with abnormal vascular structure and such abnormalities contribute to neurological impairment. Thus, new strategies to correct vascular abnormalities in brain are important for improving patient recovery and clinical outcomes.

描述（由申请人提供）：拟议研究的广泛长期目标是确定控制脑中血管形态发生的基本机制，从而确定新的合理的基于药物的策略，用于使因缺血和创伤而形成的病理血管正常化。一个平行的目标是了解负责脑中病理性血管生成的分子和细胞机制。拟议研究的工作假设是，病理性血管生成是异常血管形态发生的结果，并且可以通过药物干预实现显著改善，特别是通过适当靶向内皮细胞骨架信号通路。与病理性血管生成相关的严重缺陷，包括血流受损和血管通透性过高，可能导致缺血性损伤后的神经功能缺损。通过药物干预及时纠正血管缺损，有望改善脑缺血和创伤患者的神经功能结局。为了实现这些目标，提出了两个目标。第一个目标是：在体外，定义调节脑微血管内皮细胞（BMVECs）组装成毛细血管样结构的信号通路;并确定一组调节这些信号通路的药物，从而促进血管形态发生。在这第一个目标的关键目标是定义信号通路和效应分子，调节毛细血管结构的形成BMVECs在体外，重点是Rho GTP酶，GSK-3b，和钙蛋白酶;并确定药物，选择性地针对这些途径和效应分子在BMVECs，从而改善毛细血管结构的形成在体外。实验工具将包括逆转录病毒转导的BMVECs与特定的信号转导突变体，在体外毛细血管形态发生测定，在体外分析的细胞：细胞连接，和药理学药物，调节Rho GTP酶，GSK- 3b，和钙蛋白酶的活动。第二个目标是：在体内，在成年大脑中VEGF驱动的血管生成的小鼠模型，确定合理的药物为基础的策略，以改善病理性新生血管的结构和功能。这第二个目标的关键目标是，利用目标1中的发现，确定使异常血管结构和血管灌注正常化、使基底膜和周细胞覆盖正常化并降低脑中血管通透性过高的基于药物的策略。实验方法将涉及脑中病理性血管生成的小鼠模型，以及血管结构、灌注和高渗透性的显微镜分析。公共卫生相关性：这项研究的目的是确定调节成人大脑中新血管形成的基本机制，从而设计新的策略来纠正通常与血管发育障碍相关的严重缺陷。了解脑血管缺陷的机制很重要，因为中风和创伤性损伤会导致血管结构异常的血管形成，而这种异常会导致神经功能障碍。因此，纠正脑血管异常的新策略对于改善患者恢复和临床结局非常重要。