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Novel experimental models to study the effect of extracellular vesicles on neurons

研究细胞外囊泡对神经元影响的新实验模型

基本信息

批准号：
10508346
负责人：
Erhard Bieberich
金额：
$ 42.08万
依托单位：
UNIVERSITY OF KENTUCKY
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：
2022-08-17 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10508346
关键词：
Affect Age Aging Alzheimer&apos s Disease Alzheimer&apos s disease model Amyloid beta-Protein Apoptosis Area Astrocytes Binding Brain CASP3 gene Cell Death Cells Ceramides Cognition Communication DNA Data Dependovirus Enzymes Experimental Models Gene Expression Generations Genetic Genus Hippocampus Goals Hydrolysis Impairment In Vitro Individual Intracellular Transport Knock-out Knowledge Label Length Lipids MicroRNAs Microglia Mitochondria Modeling Monitor Mus Neurobiology Neurons Nuclear Organelles Parkinson Disease Pathogenicity Pathologic Peptides Pharmacology Physiological Pilot Projects Population Prevention Process Processed Genes Prostate Proteins Publishing RNA Regulation Reporter Satellite Viruses Signal Transduction Sphingolipids Sphingomyelinase Sphingomyelins Synapses Synaptic Transmission System Techniques Testing Tissues Transcript Up-Regulation Vesicle Virulence Factors base biological research cell type experimental study extracellular vesicles improved in vivo in vivo Model lipid transport loss of function middle age mouse model neuron development neuron loss neurotoxicity neurotransmission novel prevent response synuclein tau Proteins transcriptome sequencing uptake vesicle transport

项目摘要

Extracellular vesicles (EVs) have taken center stage in almost every area of biological research, including neurobiology. EVs are secreted vesicles that transport lipids, RNA/DNA, and proteins from a donor to a recipient cell, including pathogenic peptides and proteins such as Aβ, tau, and synuclein. Therefore, spreading of pathogenic factors by EVs in Alzheimer’s disease (AD) and Parkinson’s Disease (PD) is the focus of many recent studies. However, the physiological significance of endogenously generated EVs and mechanism of function are largely unknown. A critical knowledge gap and barrier to progress is our lack of understanding of the physiological and pathological function of EVs due to the scarcity of in vivo models. We showed that EVs secreted by astrocytes and enriched with the sphingolipid ceramide (“astrosomes”) are taken up by neurons and transported to mitochondria. We discovered that genetic deficiency of neutral sphingomyelinase 2 (nSMase2), an enzyme generating ceramide by hydrolysis of sphingomyelin, reduces astrosomes and upregulates transcript clusters associated with neuronal development and signal transduction, suggesting that astrosomes regulate neuronal function. To test the regulation of neurons by astrosomes, we propose to establish novel techniques and models for EV functional analysis. We developed mouse models with-astrocyte specific nSMase2 deficiency (loss-of- function) models and secretion of fluorescently labeled astrosomes (reporter models). We will primarily use these models to understand the significance of endogenous astrosomes for normal neuronal function. We will incorporate pilot studies to understand the function of astrosomes in (changing) physiological conditions (e.g., aging). Our overall goal is to establish novel techniques to determine the in vivo function of astrosomes. Once established, these techniques can be adapted to determine the function of EVs from other cell types (neurons, microglia, or any other tissue and cell type) as well

细胞外囊泡（EV）在生物学研究的几乎每个领域都占据了中心地位，包括神经生物学EV是分泌的囊泡，其将脂质、RNA/DNA和蛋白质从供体转运到受体细胞，包括致病肽和蛋白质，如Aβ、tau和突触核蛋白。因此，关于EV在阿尔茨海默病（AD）和帕金森病（PD）中的致病因素的研究是最近许多研究的焦点，问题研究然而，内源性产生的EV的生理意义和功能机制是大部分未知。一个关键的知识差距和进步的障碍是我们缺乏对由于缺乏体内模型，EV的生理和病理功能。我们发现，由星形胶质细胞分泌并富含鞘脂神经酰胺的EV（“星形体”）被神经元吸收并转运到线粒体。我们发现中性粒细胞的遗传缺陷鞘磷脂酶2（nSMase 2），一种通过水解鞘磷脂产生神经酰胺的酶，星形体和上调与神经元发育和信号转导相关的转录簇，表明星形体调节神经元功能。为了测试星形体对神经元的调节，我们建议建立新的技术和模型， EV功能分析。我们开发了星形胶质细胞特异性nSM酶2缺陷（缺失）小鼠模型，功能）模型和荧光标记的星形体的分泌（报告模型）。我们将主要使用这些模型，以了解正常神经元功能的内源性星形体的意义。我们将结合试点研究以了解星形体在（变化的）生理条件下的功能（例如，老化）。我们的总体目标是建立新的技术，以确定在体内的功能的星形体。一旦建立，这些技术可以适用于确定来自其他细胞类型（神经元，小胶质细胞或任何其他组织和细胞类型）