Investigating mechanisms of microvesicle release and function by GDE2 and GDE3

研究 GDE2 和 GDE3 微泡释放和功能的机制

• 批准号: 10457480
• 负责人: Reuben Elliot Levy-Myers
• 金额: $ 4.68万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-24 至 2023-08-23
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10457480
• 关键词: Abeta synthesis; Address; Affect; Alzheimer' s Disease; Alzheimer' s disease patient; Amyloid beta-Protein; Anabolism; Astrocytes; Autopsy; Biochemical; Biogenesis; Bioinformatics; Biological Markers; Biology; Brain; Cancer Biology; Cell membrane; Cell surface; Cells; Classification Scheme; Conflict (Psychology); Data; Disease; Endosomes; Environment; Enzymes; Failure; Family; Future; Grant; Health; Hippocampus (Brain); Human; In Vitro; Integral Membrane Protein; Knock-out; Label; Maintenance; Malignant Neoplasms; Measures; Mediating; Membrane; Mentors; Multivesicular Body; National Research Service Awards; Nerve Degeneration; Nervous System Physiology; Neuraxis; Neurodegenerative Disorders; Neurons; Neurosciences; Outcome Study; Pathogenicity; Pathology; Pathway interactions; Phenotype; Physiological; Physiology; Play; Primary Cell Cultures; Process; Production; Proteins; Proteomics; Publications; Publishing; RNA; Research; Research Personnel; Role; Route; Signal Transduction; Structure; Surface; Synapses; System; Techniques; Testing; Therapeutic; Tissues; Training; Vesicle; Viral; Work; biophysical properties; career; deep sequencing; delivery vehicle; exosome; experience; experimental study; extracellular; extracellular vesicles; glycerophosphodiester phosphodiesterase; human disease; insight; member; microvesicles; mouse genetics; neuron loss; neuronal survival; neuroprotection; novel; novel marker; novel therapeutic intervention; novel therapeutics; prion-like; skills; tau Proteins; vesicular release

Project Summary / Abstract Extracellular vesicles (EVs) are small membrane bound vesicles released by cells that have recently emerged as central players for many aspects of human health, including AD. However, these initial studies have raised conflicting data about whether EVs are neuroprotective or pathogenetic. EVs can be broadly divided into exosome and microvesicles (MVs), and work in other fields suggests that EV subtypes can play specialized roles. Accordingly, more work is needed to characterize subtypes of EVs in the central nervous system (CNS) and determine if these have neuroprotective or pathogenetic functions. Our lab has identified the six- transmembrane enzymes GDE2 and GDE3 as novel releasers of MVs. Preliminary and published studies ascribe GDE2 and GDE3 roles in regulating neuronal survival and indicate that GDE2 distribution and function are disrupted in Alzheimer's Disease (AD). This proposal will test the central hypothesis that GDE2 and GDE3 utilize a unique biosynthesis pathway to release MVs that have neuroprotective function, and that this release pathway is disrupted in AD. Specifically, this proposal will separately examine the MV biosynthesis pathway of GDE3 and GDE2 (Aim 1) and identify protein and RNA cargo of GDE2 and GDE3 EVs (Aim 2). Experiments in Aim 3 will characterize the disruption of GDE2 localization and MV release in AD, and asses the neuroprotective capabilities of GDE2 MVs in vitro. Successful completion of this project will characterize the biosynthesis and function of novel subtypes of EVs in the CNS and determine if they are perturbed in AD. The proposed project will provide excellent training for me in research. Specifically, I will gain new experiences in the fields of EV biology and neuroscience and gain new expertise across multiple techniques that include EV isolation and biochemical analysis, proteomics and deep sequencing, bioinformatics, mouse genetics, primary cell-culture, viral transduction, and human disease. The research environment is highly collaborative, I will have multiple opportunities to mentor and teach, and develop my scientific presentation skills though presentations and publications. Overall, this Kirschstein-NRSA grant will not only provide new insight into fundamental EV biology and explore relevance to AD, but will also prepare me for my future career as an independent investigator.

项目摘要 /摘要 细胞外囊泡（EV）是细胞释放的小膜结合囊泡，这些囊泡最近出现了 作为包括AD在内的许多方面的核心参与者。但是，这些最初的研究已经提出 关于电动汽车是神经保护性还是致病性的数据相互矛盾。电动汽车可以大致分为 外泌体和微泡（MV），在其他领域的工作表明EV子类型可以发挥专业的作用 角色。因此，需要更多的工作来表征中枢神经系统（CNS）中电动汽车的亚型 并确定这些是否具有神经保护作用或致病功能。我们的实验室已经确定了六个 跨膜酶GDE2和GDE3作为MV的新释放器。初步和发表的研究归因于 GDE2和GDE3在调节神经元存活中的作用，并表明GDE2分布和功能是 在阿尔茨海默氏病（AD）中破坏。该提案将检验GDE2和GDE3使用的中心假设 释放具有神经保护功能的MVS的独特生物合成途径，该释放途径 在广告中被破坏。具体而言，该建议将分别检查GDE3和 GDE2（AIM 1）并鉴定GDE2和GDE3 EV的蛋白质和RNA货物（AIM 2）。 AIM 3中的实验将 表征AD中GDE2定位和MV释放的破坏，并评估神经保护作用 GDE2 MVS体外的功能。该项目的成功完成将表征生物合成和 中枢神经系统中电动汽车的新型亚型的功能，并确定它们是否在AD中扰动。拟议的项目 将为我提供出色的研究培训。具体来说，我将在EV领域获得新的经验 生物学和神经科学，并在包括电动汽车隔离和的多种技术中获得新的专业知识 生化分析，蛋白质组学和深层测序，生物信息学，小鼠遗传学，原代细胞培养， 病毒转导和人类疾病。研究环境是高度协作的，我将有多个 指导和教学的机会，并发展我的科学演讲技巧，并通过演讲和 出版物。总体而言，这项Kirschstein-NRSA赠款不仅将提供有关基本EV生物学的新见解 并探索与AD的相关性，但也将为我作为独立研究者的未来职业做好准备。