The Role of CLIC4 in Angiogenesis

CLIC4 在血管生成中的作用

• 批准号: 7894662
• 负责人: JOHN C EDWARDS
• 金额: $ 38.23万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-17 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7894662
• 关键词: Acids; Address; Affect; Back; Biochemical; Biological Assay; Blood Vessels; Blood capillaries; CLIC4 gene; Caenorhabditis elegans; Capillary Endothelial Cell; Cell Proliferation; Cell fusion; Cell membrane; Cells; Cessation of life; Chloride Channels; Chloride Ion; Chlorides; Coronary Arteriosclerosis; Coupled; Data; Defect; Development; Diabetes Mellitus; Disease; Electrophoresis; Embryo; Endothelial Cells; Environment; Failure; Family; Family member; Goals; Growth; Heart Diseases; In Vitro; Individual; Intracellular Membranes; Ion Channel; Ions; Lead; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Measures; Mediator of activation protein; Membrane; Membrane Fusion; Membrane Protein Traffic; Migraine; Modification; Molecular; Morphogenesis; Mus; Nephrotic Syndrome; Pathway interactions; Permeability; Physiological; Pilot Projects; Pinocytosis; Plasma; Play; Point Mutation; Prevention; Process; Property; Protein Family; Proteins; Proton Pump; Reagent; Recombinants; Regulation; Risk; Role; Science; Signal Transduction; Staging; Stimulus; Stroke; Structure-Activity Relationship; Sudden infant death syndrome; Swelling; Tubular formation; Vacuole; Vesicle; angiogenesis; base; capillary; clinically relevant; common treatment; design; extracellular; human disease; in vivo; inhibitor/antagonist; insight; membrane activity; novel strategies; prenatal; public health relevance; response; retinal angiogenesis; two-dimensional

DESCRIPTION (provided by applicant): Angiogenesis, the process that new blood vessels form from existing vessels, is a critical process in development and response to environment and disease. Alterations in vascular development and angiogenesis play important roles in many diverse disease including diabetes, coronary artery disease, stroke, and cancer. More complete understanding of the mechanisms leading to appropriately controlled angiogenesis may lead to new approaches for prevention and/or treatment of these common debilitating diseases. Capillary tubules form from endothelial cells by a process called "cell-hollowing" tubulogenesis, which involves formation and fusion of intracellular vesicles to form a large intracellular vacuole that eventually becomes the extracellular lumen of the tubule. CLIC proteins are a family of chloride channel proteins which largely reside in intracellular membranes. Chloride permeability has long been recognized to play a role in intracellular membrane traffic but the specific proteins responsible for individual chloride conductance of intracellular membranes is still largely unknown. Recently, a CLIC protein has been shown to be essential for a specific cell-hollowing tubulogenic process in C. elegans and the family member CLIC4 has been implicated in tubulogenesis in vitro of mammalian endothelial cells. We have recently generated and characterized mice in which the gene for CLIC4 has been disrupted. Preliminary data from these mice provide startling and exciting new evidence that CLIC4 is indeed involved in angiogenesis. Our hypothesis is that CLIC4 plays a critical role in angiogenesis, perhaps by providing the chloride conductance necessary for the orderly membrane traffic and membrane fusion steps involved in cell-hollowing tubulogenesis of endothelial cells. We propose to: 1) Study CLIC4 (-/-) mice for abnormalities in vascular development and response to angiogenic stimuli; 2) Identify sequences and motifs within CLIC4 that are necessary for its function in cell-hollowing tubulogenesis; and 3) Investigate transport properties of membranes derived from cells undergoing tubulogenesis for an activity attributable to CLIC4 and explore how that effects tubulogenesis. The proposal thus is a hypothesis-driven basis science project that uses novel approaches and reagents to address a recently-identified key step in angiogenesis about which little is currently known mechanistically. The results will have clear clinical relevance to the many diseases in which angiogenesis plays a part. PUBLIC HEALTH RELEVANCE: Growth and regulation of new blood vessels is important in normal development and critical in several important diseases such as cancer, heart disease and stroke. This proposal focuses on newly discovered key role for an intracellular chloride channel in development of new blood vessels. The results of the proposed studies will provide new insight into these common, important disease processes and may open new avenues for prevention and treatment.

描述（由申请人提供）：血管生成是从现有血管形成新血管的过程，是发育和对环境和疾病反应的关键过程。 血管发育和血管生成的改变在包括糖尿病、冠状动脉疾病、中风和癌症在内的许多不同疾病中起重要作用。更完整的了解机制，导致适当控制血管生成可能会导致新的方法来预防和/或治疗这些常见的衰弱性疾病。毛细血管小管通过称为“细胞中空化”小管发生的过程从内皮细胞形成，该过程涉及细胞内囊泡的形成和融合以形成大的细胞内空泡，其最终成为小管的细胞外腔。CLIC蛋白是主要存在于细胞内膜中的氯离子通道蛋白家族。 氯渗透性长期以来被认为在细胞内膜运输中起作用，但负责细胞内膜的个体氯电导的特异性蛋白质仍然在很大程度上是未知的。最近，CLIC蛋白已被证明是必不可少的一个特定的细胞中空微管的过程中，在C。elegans和家族成员CLIC 4与哺乳动物内皮细胞的体外小管形成有关。 我们最近已经产生并表征了CLIC 4基因被破坏的小鼠。 来自这些小鼠的初步数据提供了令人惊讶和令人兴奋的新证据，表明CLIC 4确实参与了血管生成。 我们的假设是CLIC 4在血管生成中起着关键作用，可能是通过提供有序的膜运输和膜融合步骤所必需的氯离子电导，这些步骤涉及内皮细胞的细胞中空化小管发生。 我们建议：1）研究CLIC 4（-/-）小鼠的血管发育异常和对血管生成刺激的反应; 2）鉴定CLIC 4内的序列和基序，所述序列和基序是其在细胞中空化小管发生中的功能所必需的;和3）研究源自经历小管发生的细胞的膜的转运特性，以获得归因于CLIC 4的活性，并探索其如何影响小管发生。因此，该提案是一个假设驱动的基础科学项目，使用新的方法和试剂来解决最近确定的血管生成关键步骤，目前对该步骤的机制知之甚少。这些结果将对血管生成起作用的许多疾病具有明确的临床相关性。公共卫生相关性：新血管的生长和调节在正常发育中是重要的，并且在几种重要疾病如癌症、心脏病和中风中是关键的。该提案的重点是新发现的细胞内氯离子通道在新血管发育中的关键作用。拟议研究的结果将为这些常见的重要疾病过程提供新的见解，并可能为预防和治疗开辟新的途径。