Polarized Protein Trafficking and Angiogenesis

极化蛋白运输和血管生成

基本信息

批准号：
10539327
负责人：
Erich J Kushner
金额：
$ 36.06万
依托单位：
UNIVERSITY OF DENVER (COLORADO SEMINARY)
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10539327
关键词：
3-Dimensional Ablation Alleles Apical Architecture Binding Biochemical Biochemistry Biogenesis Blood Blood Vessels CRISPR imaging Cardiovascular Diseases Cell Culture System Cell membrane Cells Chemosensitization Clustered Regularly Interspaced Short Palindromic Repeats Complex Data Development Developmental Process Distant Docking Embryo Endothelial Cells Endothelium Event Excision Generations Genes Genetic Genetic Transcription Goals Guanine Guanine Nucleotide Exchange Factors Guanosine Triphosphate Phosphohydrolases Homeostasis Image Imaging Techniques Immune response In Vitro Investigation Label Laboratories Maps Mediating Membrane Microscopy Molecular Morphogenesis Movement Mutation Names Nutrient Oxygen Pathway interactions Pattern Post-Translational Protein Processing Primary Cell Cultures Process Protein Engineering Proteins Reporter Resolution Role Signal Transduction Surface Tertiary Protein Structure Testing Time Tissues Transgenic Organisms Vesicle Work Zebrafish angiogenesis apical membrane blood vessel development body system density in vivo insight luminal membrane mutant novel overexpression oxygen transport podocalyxin programs protein transport rab GTP-Binding Proteins synaptotagmin trafficking tumor growth wasting wound healing

项目摘要

SUMMARY Blood vessels carry oxygen and nutrients and are vital to organismic viability and continued homeostasis. Angiogenesis, or the formation of new blood vessels from pre-existing ones, is the predominant developmental process by which blood vessel network density is regulated. During angiogenic development, endothelial cells create a hollow cavity called a lumen, providing a continuous conduit for blood to reach distant tissues. The mechanisms underpinning the morphodynamic changes in endothelial architecture and signaling leading to vascular lumen formation, or tubulogenesis, are incompletely understood. In this proposal we will investigate a protein called synaptotagmin-like protein 2 (sytl2) that we believe is responsible for deﬁning the luminal surface by directing protein transport to the apical membrane during blood vessel development. Our preliminary data suggests that sytl2 deﬁnes the apical membrane and tethers Rab GTPase proteins for delivery of vesicular cargo, such as podocalyxin. In aim 1, we will characterize the role of sytl2a during vascular lumen formation in developing zebraﬁsh embryos using a combination of live-imaging and CRISPR-based mutant generation. In aim 2, we will comprehensively demonstrate that sylt2 works in combination with the GTPase Rab35 to deliver podocalyxin to the apical plasma membrane during lumenogenesis in vitro. In aim 3, we will further characterized how sytl2a interacts with Rab35 to deliver Podocalyxin using generation of new zebraﬁsh reporter lines and compound mutants in vivo. How blood vessel lumen formation is regulated is still a major question in the ﬁeld, this proposal will provide novel insight into critical mechanisms orchestrating this process.

概括血管携带氧气和营养物质，对于有机体的活力和持续的体内平衡至关重要。血管生成，或从现有血管形成新血管，是主要的发育过程在血管生成发育过程中，内皮细胞调节血管网络密度的过程。创建一个称为管腔的空腔，为血液到达远处组织提供连续的管道。支持内皮结构和信号传导形态动力学变化的机制血管腔形成或肾小管发生尚不完全清楚，在本提案中，我们将研究一个问题。称为突触结合蛋白样蛋白 2 (sytl2) 的蛋白质，我们认为它负责定义管腔表面我们的初步数据是在血管发育过程中将蛋白质转运至顶膜。表明 sytl2 定义了顶膜并束缚 Rab GTPase 蛋白以递送囊泡在目标 1 中，我们将描述 sytl2a 在血管腔形成过程中的作用。使用实时成像和基于 CRISPR 的突变体生成相结合来开发斑马鱼胚胎。目标2，我们将全面证明sylt2与GTPase Rab35结合发挥作用在体外腔发生过程中足萼蛋白到顶端质膜的作用在目标 3 中，我们将进一步。描述了 sytl2a 如何与 Rab35 相互作用以利用新一代斑马鱼传递 Podocalyxin 体内报告系和复合突变体如何调节血管腔形成仍然是一个主要问题。问题，该提案将为协调这一过程的关键机制提供新颖的见解。