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In vitro and in vivo Signaling Mechanisms during Endothelial Tube Formation

内皮管形成过程中的体外和体内信号传导机制

基本信息

批准号：
8725185
负责人：
Jeffrey J Essner
金额：
$ 27.04万
依托单位：
IOWA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8725185
关键词：
Actin-Binding Protein Address Adherence Adherens Junction Affect Aorta Apical Biological Blood Vessels Cadherins Caliber Cell Communication Cell Polarity Cells Chimeric Proteins Conflict (Psychology)Coupled Coupling Development Disease Dorsal Embryo Endothelial Cells Equilibrium Future Gene Expression Gene Transfer Techniques Generations Genes Genome Goals Grant Heart Diseases Human Human Genome In Vitro Injection of therapeutic agent Ischemia Knowledge Malignant Neoplasms Mediating Membrane Molecular Monomeric GTP-Binding Proteins Mus Oligonucleotides Optics Pathway interactions Phenotype Process Protein Serine/Threonine Phosphatase Proteins Publishing Regulation Research Role Signal Transduction Structure Surface Techniques Testing Therapeutic Intervention Tight Junctions Transgenic Animals Transgenic Organisms Transport Vesicles Tube Tumor Angiogenesis Umbilical vein Vacuole Vascular System Vesicle Work Zebrafish angiogenesis apical membrane base comparative experience in vivo in vivo imaging innovation insight luminal membrane membrane biogenesis novel novel therapeutic intervention novel therapeutics protein protein interaction research study selective expression therapeutic development trafficking treatment strategy tumor progression vascular bed vasculogenesis

项目摘要

DESCRIPTION (provided by applicant): Our long-term goal is to understand the cellular mechanisms that regulate vesicle trafficking during endothelial tube formation. Our objective is to define critical genes involved in the formation of the apical membrane and the coordination between vesicle trafficking and the remodeling of cellular junctions during endothelial tube formation. Our principle hypothesis is that endothelial tube formation is tightly coupled to adherence and tight junction remodeling and that these apical junctions direct vesicle trafficking during formation of the apical membrane in vitro and in vivo. In support of this hypothesis, we have identified a number of proteins involved in cellular junctions and vesicle trafficking and demonstrated that these genes are required for endothelial tube formation in human umbilical vein endothelial cells in culture and during embryonic angiogenesis in zebrafish. We have chosen to work with zebrafish because of the combined advantages of a well-defined genome that relates to the human genome, the ability to efficiently knockdown genes and produce transgenic animals, and the optical clarity of the embryo that allows a detailed examination of tube formation on the single cell level in vivo. Complementary to this approach, we follow the same processes biochemically in vitro. We provide preliminary evidence suggesting a close coupling of vesicle trafficking and apical junctional remodeling. Our team has extensive experience with the generation of transgenic zebrafish using transposon-mediated transgenesis techniques, the injection of antisense morpholino oligonucleotides, and the examination of protein-protein interactions in vitro. We have taken an innovative and comparative approach to this problem by combining in vitro and in vivo experiments. We use transgenic zebrafish and human endothelial cells that express fluorescently tagged proteins to answer fundamental cell biological questions regarding endothelial tube formation. At the end of the granting period, we will have defined novel mechanisms that function during endothelial tube formation. This understanding is critical to the development of new therapeutic strategies to modulate of endothelial tube formation and diameter. This ability would have direct implications for the treatment of heart disease, ischemia, and cancer.

描述（由申请人提供）：我们的长期目标是了解内皮管形成过程中调节囊泡运输的细胞机制。我们的目标是确定参与顶端膜形成的关键基因，以及在内皮管形成过程中囊泡运输和细胞连接重塑之间的协调。我们的原则假设是，内皮管的形成是紧密耦合的粘附和紧密连接重塑，这些顶端连接直接囊泡运输过程中的顶端膜在体外和体内形成。为了支持这一假设，我们已经确定了一些蛋白质参与细胞连接和囊泡运输，并证明这些基因是所需的内皮管形成在培养的人脐静脉内皮细胞和胚胎血管生成过程中的斑马鱼。我们选择用斑马鱼进行研究，是因为斑马鱼具有与人类基因组相关的明确定义的基因组的综合优势，能够有效地敲除基因并产生转基因动物，以及胚胎的光学清晰度，允许在体内单细胞水平上详细检查管形成。作为这种方法的补充，我们在体外遵循相同的生化过程。我们提供的初步证据表明，囊泡贩运和顶端连接重塑的密切耦合。我们的团队在使用转座子介导的转基因技术、注射反义吗啉代寡核苷酸以及体外蛋白质-蛋白质相互作用的检查来产生转基因斑马鱼方面具有丰富的经验。我们已经采取了一种创新的和比较的方法来解决这个问题，结合在体外和体内实验。我们使用转基因斑马鱼和人类内皮细胞，表达荧光标记的蛋白质来回答有关内皮管形成的基本细胞生物学问题。在授权期结束时，我们将确定在内皮管形成过程中发挥作用的新机制。这种理解对于开发新的治疗策略以调节内皮管形成和直径至关重要。这种能力将对心脏病、局部缺血和癌症的治疗产生直接影响。