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养分，对有机生存能力和继续稳态至关重要。血管生成或先前存在的血管形成新血管，是主要发育调节血管网络密度的过程。在血管生成期间，内皮细胞创建一个称为管腔的空心腔，为血液提供连续的管道，以使血液到达远处的组织。支撑内皮体系结构和信号传导形态动力学变化的机制导致血管腔的形成或微管发生不完全理解。在此提案中，我们将调查蛋白质称为突触类蛋白样蛋白2（SYTL2），我们认为这是负责定义腔表面的蛋白质通过将蛋白质转运到血管发育过程中。我们的初步数据建议SYTL2定义顶端膜和Tethers RAB GTPase蛋白用于递送水泡货物，例如足球素。在AIM 1中，我们将表征Sytl2a在血管管腔形成中的作用使用现场成像和基于CRISPR的突变体产生的组合，开发斑马鱼胚胎。在 AIM 2，我们将全面证明SYLT2与GTPASE RAB35结合使用以交付在体外发光期间，足体质质膜上的足球素。在AIM 3中，我们将进一步表征了Sytl2a与Rab35相互作用的方式以使用新的斑马鱼的产生来递送足豆激素记者线和体内复合突变体。如何调节血管管腔的形成仍然是主要的该提案中的问题将在策划这一过程的关键机制方面提供新的见解。