Molecular Mechanisms Underlying Endothelial Weibel-Palade Body Biogenesis and Exocytosis

内皮 Weibel-Palade 体生物发生和胞吐作用的分子机制

• 批准号: 10212750
• 负责人: Anish Vaibhav Sharda
• 金额: $ 16.5万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10212750
• 关键词: Adaptor Signaling Protein; Advisory Committees; Affect; Albinism; Binding; Biogenesis; Blood; Blood Cells; Blood Coagulation Disorders; Blood Platelets; Cardiovascular system; Cell membrane; Cells; Clinical Research; Coagulation Process; Complex; Cytoplasmic Granules; Data; Defect; Dependence; Disease; Docking; Endosomes; Endothelial Cells; Endothelium; Environment; Exocytosis; Fellowship; Five-Year Plans; Genes; Goals; Growth; Hematology; Hemorrhage; Hemostatic Agents; Hemostatic function; Hermanski-Pudlak Syndrome; Human; Immunoprecipitation; Impairment; Individual; Israel; Lysosomes; Mediating; Medical center; Melanosomes; Membrane; Membrane Protein Traffic; Mentors; Mentorship; Molecular; Molecular Weight; Morbidity - disease rate; Neoplasm Metastasis; Organelles; Pathway interactions; Plasma; Play; Postdoctoral Fellow; Process; Proteins; Regulation; Reporting; Research Personnel; Role; SNAP receptor; SNAP23 gene; Scientist; Secondary to; Secretory Vesicles; Skin Pigmentation; Thrombosis; Training; Vesicle; Weibel-Palade Bodies; Yeasts; angiogenesis; cancer cell; cardiovascular risk factor; career; cellubrevin; experimental study; fluidity; in vivo; innovation; live cell microscopy; mouse model; platelet function; protein complex; protein transport; rab GTP-Binding Proteins; receptor; release factor; syntaxin 3; trafficking; trans-Golgi Network; von Willebrand Disease; von Willebrand Factor

Project Summary/Abstract Endothelium plays a vital role in hemostasis and thrombosis. It is essential to maintain blood fluidity, but it also expresses and releases numerous “factors” that regulate blood cell activation and coagulation. Among these, von Willebrand factor (vWF) is an essential plasma hemostatic factor released by the endothelium. Deficiency of mature vWF is the most common bleeding disorder in humans. On the other hand, elevated plasma levels of vWF, or abnormal concentrations of its high-molecular weight multimers, is associated with increased risk of cardiovascular morbidity. As such, understanding of the molecular mechanisms that underlie regulated release of vWF have broad implications in hemostasis and thrombosis. One factor that plays a crucial role in normal vWF exocytosis from endothelial cells is unimpaired biogenesis of its storage granules, the Weibel-Palade bodies (WPBs), which is a highly complex process that remains poorly characterized. We have previously shown that deficiency of biogenesis of lysosome-related organelle complex 2 (BLOC-2) results in impaired vWF exocytosis. Since, BLOC-2, and other related trafficking proteins, are essential for biogenesis of lysosomal-related organelles, a group of specialized granules that includes platelet dense granules and melanosomes, it seems plausible that this protein complex is also required for biogenesis of WPB. We hypothesize that BLOC-2-mediated endosomal trafficking is critical for biogenesis of WPB. We further postulate that the exocyst complex plays an essential role in WPB trafficking by 1) interacting with BLOC-2 in its role in endosomal trafficking 2) and regulating soluble NSF adaptor protein receptor (SNARE)-mediated fusion of WPBs at the plasma membrane. In Aim 1, we will characterize cargo trafficking from the endosomes to the maturing WPBs and the dependence of this pathway on BLOC-2. In Aim 2, we will evaluate the role of the exocyst complex in BLOC-2-dependent endosomal trafficking and characterize the core trafficking machinery involved in this pathway. And, finally, in Aim 3, we will evaluate the regulatory function of the exocyst complex in SNARE-mediated fusion of WPBs at the plasma membrane. The applicant, Dr. Anish Sharda, previously completed clinical and research fellowship in hematology and is currently pursuing a post-doctoral fellowship under the mentorship of Drs. Robert Flaumenhaft and Bruce Furie, who will serve as primary mentor and co-mentor, respectively. The Division of Hemostasis and Thrombosis at Beth Israel Deaconess Medical Center has a distinguished track record of scientific innovation and mentorship, and will provide an excellent environment for the applicant in pursuit of his career goals. The advisory committee of exceptional scientists that Dr. Sharda has established will bring diverse intellectual expertise to his training and scientific growth. Dr. Sharda is well-qualified to execute the proposed experiments and has presented a comprehensive five-year plan to meet his goal of becoming an independent researcher.

项目摘要/摘要 内皮在止血和thombosis中起着至关重要的作用。 表达并释放了许多“因素”，使血细胞激活和凝结。 Von Willbrand因子（VWF）是内皮释放的特别血浆止血因子。 另一方面，成熟的VWF是人类最常见的出血障碍。 VWF或其高分子重量多聚体的异常收缩，被拒绝，增加了 心血管疾病的理解。 VWF的止血和血栓形成具有广泛的影响。 内皮细胞的VWF胞吐作用是储存颗粒的Isunimpaired Biogeneseres，Weibel-Palade 身体（WPB），这是我们以前具有的高度综合过程过程 表明与溶酶体相关细胞器复合物2（BLOC-2）的生物发生不足导致受损 VWF胞吐作用。 溶酶体相关的细胞器，一组专门的颗粒，包括血小板密集颗粒和 黑色素体似乎是蛋白质复合物的合理，也用于WPB的生物发生 假设BLOC-2介导的内体运输对于WPB的生物发生至关重要。 假设外囊综合体在WPB运输中起着至关重要的作用1）与BLOC-2相互作用相互作用 它在内体贩运中的作用2）和调节可溶性NSF适配器蛋白受体（SNARE）介导的 WPB在质膜上的融合。 对WPB的含量和该途径对AIM 2的依赖性 依赖BLOC-2依赖性内体贩运的外囊综合体并表征了核心贩运 这项途径涉及的机械。最后，在AIM 3中，我们将评估 在质膜上WPB的网罗介导的融合中的外囊复合物。 申请人Anish Sharda博士先前完成了血液学的临床和研究奖学金 目前，正在博士的指导下追求博士后奖学金 布鲁斯·弗里（Bruce Furie）将担任主要导师和同事的尊敬者。 贝丝以色列女执事医疗中心的血栓形成具有杰出的科学创新记录 和指导，并将为申请人追求自己的职业目标提供一个绝佳的环境 异常咨询委员会塔尔达（Tharda）已建立的咨询委员会将带来多元化 他的培训和科学成长的专业知识。 并提出了一项全面的五年计划，以实现他为独立研究人员提供的目标。