Exploring exosomes in neurodevelopmental and neuropsychiatric diseases using brain organoids

使用脑类器官探索外泌体在神经发育和神经精神疾病中的作用

基本信息

批准号：
10741385
负责人：
Dilek Colak
金额：
$ 46.39万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10741385
关键词：
ASD patient Affect Alzheimer&apos s Disease Astrocytes Autopsy Biochemical Markers Biogenesis Bioinformatics Brain Cell Communication Cell Count Cells Central Nervous System Cerebrum Classification Collection Communication Culture Media Data Databases Development Disease Endosomes Etiology Goals Harvest Health Heterogeneity Human In Vitro Inflammatory Lipids Literature Malignant Neoplasms Mammalian Cell Mediating Membrane Mental disorders Methyl-CpG-Binding Protein 2 MicroRNAs Microglia Modeling Modification Multiple Sclerosis Nerve Degeneration Neurocognitive Neurodegenerative Disorders Neurodevelopmental Disorder Neuroglia Neuronal Differentiation Neurons Nucleic Acids Oligodendroglia Organoids Parkinson Disease Pathogenesis Pathway Analysis Patients Phenotype Process Protein Analysis Proteins Proteome Protocols documentation Psyche structure RNA Reporting Research Rett Syndrome Role Signal Transduction Synaptic plasticity System Techniques Time Tissues Translational Regulation Transmission Electron Microscopy Ultracentrifugation Vesicle Western Blotting autism spectrum disorder brain cell cell type comparison control differentiation protocol directed differentiation exosome experimental study extracellular extracellular vesicles human stem cells in vivo induced pluripotent stem cell infancy insight intercellular communication knock-down liquid chromatography mass spectrometry mouse model nanoparticle nanovesicle neural neural circuit neurogenesis neuroinflammation neuropathology neuropsychiatric disorder next generation sequencing novel nucleic acid purification stem cell model synaptogenesis transmission process

项目摘要

PROJECT SUMMARY Exosomes are small extracellular vesicles that mediate intercellular signaling in the brain without requiring direct contact between cells. These vesicles are enriched with miRNAs, proteins, and lipids. Current evidence for exosome signaling in the brain points toward their role in translational regulation, neurogenesis, synaptic plasticity, and neuroinflammation, all of which have been implicated in several neurodevelopmental diseases. However, changes in exosome signaling in mental disorders have yet to be explored. Exosomes have been isolated from nearly all mammalian cell types, including cells in the central nervous system such as neurons, astrocytes, oligodendrocytes, and microglia. Neurons and glia release exosomes in vitro and in vivo. Trophic support from glial cells to neurons is thought to be conveyed by exosomes. Exosome-mediated intercellular signaling has been implicated in neurodegenerative diseases such as ALS, Parkinson’s disease, multiple sclerosis, and Alzheimer’s Disease as well as neurodevelopmental disorders. More specifically, altered neuronal exosome signaling has been implicated in Rett syndrome, a form of Autism Spectrum Disorder (ASD). However, the role of exosome signaling in ASD and other mental disorders remains to be fully understood. The goal of this application is to explore whether exosome content and numbers are altered in ASD. We have isolated exosomes from healthy control and patient brain organoid cultures by differential ultracentrifugation and evaluated purified exosomes using transmission electron microscopy, western blotting for biochemical markers, and nanoparticle characterization system. In purified exosome fractions, we will first assess exosome proteomes and nucleic acid profiles that will be compared between control and patient groups. In addition to content, there is also evidence suggesting that exosome quantity is altered in disease states. To explore the role of neural exosomes in ASD, we propose to determine 1) whether the proteome of exosomes isolated from ASD patient organoids differ from exosomes derived from healthy control organoids (e.g., by comparing protein profiles with Tandem mass tag [TMT] liquid chromatography [LC] mass spectrometry), 2) whether nucleic acid content in exosomes purified from ASD patient organoids is altered in comparison to exosomes from control (e.g., by comparing RNA/miRNA profiles with next-generation sequencing followed by bioinformatics), and 3) whether the quantity of exosomes released from ASD brain cells is altered compared to control cells (e.g., by assessing the quantity of exosomes in 2D neuronal or glial culture media using nanosight tracking analysis). Exosome analysis from organoids and 2D cultures will serve as parallel strategies to delineate neural exosome content in health and disease. Our studies have the potential to provide novel insights into the etiology of ASD. Identifying alterations in amount or content of brain exosomes in ASD will reveal underappreciated modifications in cellular communication during mental disease states.

项目摘要外泌体是细胞外蔬菜，可介导大脑中的细胞间信号传导而无需细胞之间的直接接触。这些蔬菜富含miRNA，蛋白质和脂质。当前的证据对于大脑中的外泌体信号传导指向其在翻译调节，神经发生，突触中的作用可塑性和神经炎症，所有这些都暗示了几种神经发育疾病。但是，精神障碍中外泌体信号的变化尚未探索。外泌体已从几乎所有哺乳动物细胞类型中分离出来，包括中枢神经中的细胞神经元，星形胶质细胞，少突胶质细胞和小胶质细胞等系统。神经元和胶质释放外泌体体内和体内。人们认为来自神经胶质细胞到神经元的营养支持是由外泌体传达的。外部介导的细胞间信号已在神经退行性疾病（例如ALS）中隐含帕金森氏病，多发性硬化症和阿尔茨海默氏病以及神经发育疾病。更具体地说，在Rett综合征中暗示了神经元外部信号传导，一种形式自闭症谱系障碍（ASD）。但是，外泌体信号在ASD和其他心理中的作用疾病仍有待完全理解。该应用程序的目的是探索是否外部内容和数字在ASD中发生了变化。我们有健康对照和患者大脑的孤立外泌体通过差分超速离心的器官培养物，并使用传播评估了纯化的外泌体电子显微镜，生化标记的蛋白质印迹和纳米颗粒表征系统。纯化的外泌体分数，我们将首先评估将是在对照组和患者组之间进行比较。除内容外，还有证据表明疾病状态中的外泌体数量发生了变化。探索神经外泌体在ASD中的作用，我们建议确定1）从ASD患者类器官中分离出的外泌体的蛋白质组是否不同于外泌体源自健康对照器官（例如，通过将蛋白质曲线与串联质量标签进行比较[TMT]液体色谱[LC]质谱法），2）外泌体中的核酸含量是否从ASD纯化与对照组的外泌体相比，患者类器官发生了变化（例如，通过比较RNA/miRNA轮廓下一代测序，然后是生物信息学），3）外泌体数量是否存在与对照细胞相比，从ASD脑细胞释放的改变（例如，通过评估外泌体的数量在2D神经元或神经胶质培养基中，使用纳米视跟踪分析）。从器官的外泌体分析和2D文化将作为描述健康中神经外泌体含量的并行策略疾病。我们的研究有可能提供对ASD病因的新见解。识别 ASD中大脑外泌体的数量或含量的改变将揭示未充分的修改在精神疾病状态下的细胞通信中。