The Role of Vitronectin in Neuron-Microglia Interactions in the Context of Social Stress

玻连蛋白在社会压力背景下神经元-小胶质细胞相互作用中的作用

基本信息

批准号：
10827550
负责人：
Daniela Franco Hernandez
金额：
$ 3.83万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
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：
2023-09-20 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10827550
关键词：
Animals Antigen-Antibody Complex Anxiety Area Atrophic Automobile Driving Autopsy Baltimore Behavior Behavioral Brain CRISPR/Cas technology Cd68 Cell Death Cell physiology Cells Cellular Structures Central Nervous System Chronic Chronic stress Clinical Committee Members Communication Communities Complement Development Dopamine Receptor Environmental Risk Factor Exhibits Exposure to Extracellular Matrix Extracellular Matrix Proteins Fellowship Female Foundations Future Generations Glycoproteins Goals Image Analysis Immune system Immunologic Surveillance Individual Inflammation Label Learning Major Depressive Disorder Maps Maryland Measures Mediating Mediator Membrane Proteins Mental Health Mentors Microglia Microscopy Molecular Morphology Motivation Mus Myelogenous Myeloid Cells Neuroimmune Neuronal Dysfunction Neurons Neurosciences Nucleus Accumbens Outcome Patients Peripheral Phase Play Postdoctoral Fellow Pre-Clinical Model Process Property Proteomics Research Research Personnel Research Project Grants Rewards Risk Factors Rodent Role Social Interaction Stress Sucrose Synapses System Technology Testing Tissues Training Transgenic Organisms Universities Viral Viral Vector Vitronectin Work behavioral outcome brain reward regions career cytokine design differential expression early life stress experience gene network immunoreactivity interest knock-down lens male medical schools migration motivated behavior negative affect neuropsychiatric disorder novel therapeutics post-doctoral training post-traumatic stress pre-clinical research preference prevent single-cell RNA sequencing skills social defeat social stress stress reduction stress related disorder transcriptome sequencing

Animals Antigen-Antibody Complex Anxiety Area Atrophic Automobile Driving Autopsy Baltimore Behavior Behavioral Brain CRISPR/Cas technology Cd68 Cell Death Cell physiology Cells Cellular Structures Central Nervous System Chronic Chronic stress Clinical Committee Members Communication Communities Complement Development Dopamine Receptor Environmental Risk Factor Exhibits Exposure to Extracellular Matrix Extracellular Matrix Proteins Fellowship Female Foundations Future Generations Glycoproteins Goals Image Analysis Immune system Immunologic Surveillance Individual Inflammation Label Learning Major Depressive Disorder Maps Maryland Measures Mediating Mediator Membrane Proteins Mental Health Mentors Microglia Microscopy Molecular Morphology Motivation Mus Myelogenous Myeloid Cells Neuroimmune Neuronal Dysfunction Neurons Neurosciences Nucleus Accumbens Outcome Patients Peripheral Phase Play Postdoctoral Fellow Pre-Clinical Model Process Property Proteomics Research Research Personnel Research Project Grants Rewards Risk Factors Rodent Role Social Interaction Stress Sucrose Synapses System Technology Testing Tissues Training Transgenic Organisms Universities Viral Viral Vector Vitronectin Work behavioral outcome brain reward regions career cytokine design differential expression early life stress experience gene network immunoreactivity interest knock-down lens male medical schools migration motivated behavior negative affect neuropsychiatric disorder novel therapeutics post-doctoral training post-traumatic stress pre-clinical research preference prevent single-cell RNA sequencing skills social defeat social stress stress reduction stress related disorder transcriptome sequencing

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项目摘要

Project Summary Exposure to chronic stress is a known risk factor for neuropsychiatric disease and yields structural and molecular changes to the brain and immune system. The nucleus accumbens (NAc), a hub for integrating reward and motivation, exhibits molecular and cellular alterations that are found in postmortem tissue of patients with stress- related disorders, such as major depressive disorder, and drive motivational deficits in rodents. Specifically, dendritic atrophy of dopamine receptor type 1 (D1), but not type 2 (D2) expressing medium spiny neurons (MSNs) is necessary and sufficient for stress-induced negative behavioral outcomes. Emerging evidence implicates microglia as potential mediators of neuronal atrophy and motivational deficits after social stress. Since microglia can interact with neurons and facilitate neuronal dysfunction and cell death, they are prime candidates for investigating the relationship between chronic stress and D1-MSN atrophy. Recent work from the lab shows that chronic social defeat stress (CSDS) reduced overall D1-MSN and microglia contact, but not D2 MSNs, and analyses of individual microglia within the D1-MSN microenvironment revealed reductions in microglia complexity in animals that exhibited negative affective behavior after CSDS. Thus, while it is evident that CSDS alters D1-MSN-microglia contact and morphology, the molecular mechanisms driving these cell-subtype specific, stress-induced changes in the NAc microenvironment remain unclear. Preliminary RNA-seq analysis from the lab points to vitronectin, an extracellular matrix substrate, as a promising molecular messenger mediating D1- MSN and microglia interactions because it plays a driving role in a network of genes altered by chronic stress and is specifically differentially expressed in D1-MSNs. In this proposal, I will virally knock down vitronectin expression in D1-MSNs using a Cre-dependent CRISPR/Cas9 construct in mice before subjecting them to either CSDS or Chronic Witness Defeat Stress (CWDS) to interrogate the role of vitronectin expression in D1-MSN- microglia interactions, D1-MSN and microglia morphology, and stress-induced behavioral outcomes. It is hypothesized that knocking down vitronectin expression in D1-MSNs will rescue D1-MSN stress-induced dendritic atrophy, restore D1-MSN-microglia contact, and prevent negative affective behavior. Understanding the molecular mechanisms driving altered stress-induced behavioral states can shed light on novel therapeutics designed to protect and/or treat the deleterious effects of chronic stress. The proposed training at the University of Maryland Baltimore, School of Medicine will facilitate my transition to a postdoctoral fellowship and allow me to continue to research stress-related neuropsychiatric disease by characterizing neuron, microglia, and extracellular matrix relationships in the context of stress exposure. With the support of my excellent scientific community comprising of my mentor, thesis committee members, collaborators, and neuroscience and Meyerhoff communities, I am excited to complete the technical and career goals outlined in the F99/K00 phases of my proposal and working and growing into the role of an independent investigator in neuroscience.

项目摘要暴露于慢性应激是神经精神疾病的已知危险因素，并产生结构和分子变化大脑和免疫系统。伏隔核（NAC），用于整合奖励和动机，表现出分子和细胞的改变，这些变化在死后组织中发现的胁迫 - 相关疾病，例如重度抑郁症，并驱动啮齿动物的动机缺陷。具体来说，多巴胺受体1型（D1）的树突状萎缩，但不表达培养基神经元2（D2）（MSN）对于压力诱导的负面行为结果是必要的，并且足够。新兴证据暗示小胶质细胞是社会压力后神经元萎缩和动机缺陷的潜在介体。自从小胶质细胞可以与神经元相互作用，并促进神经元功能障碍和细胞死亡，它们是主要候选者用于研究慢性应激与D1-MSN萎缩之间的关系。实验室的最新工作显示长期的社会失败压力（CSD）减少了总体D1-MSN和小胶质细胞接触，但不是D2 MSN，以及 D1-MSN微环境中单个小胶质细胞的分析显示，小胶质细胞降低 CSD后表现出负面情感行为的动物的复杂性。因此，尽管CSD很明显改变D1-MSN-Microglia接触和形态，分子机制驱动这些细胞型特异性，应力诱导的NAC微环境变化尚不清楚。初步的RNA-seq分析实验室指向玻染蛋白，一种细胞外基质底物，是一种有希望的分子信使介导D1- MSN和小胶质细胞相互作用，因为它在因慢性应激改变的基因网络中起驱动作用并在D1-MSN中特别差异表达。在此提案中，我将病毒击倒玻璃体素使用CRE依赖性CRISPR/CAS9在小鼠中表达在D1-MSN中的表达，然后再对其进行表达 CSD或慢性证人击败压力（CWDS），以审问玻璃体表达在D1-MSN-中的作用小胶质细胞相互作用，D1-MSN和小胶质细胞形态以及压力诱导的行为结果。这是假设击倒D1-MSN中的玻璃体表达将挽救D1-MSN应力诱导的树突状萎缩，恢复D1-MSN-microglia接触，并预防负面情感行为。理解驱动压力引起的行为状态的分子机制可以揭示新的治疗剂旨在保护和/或治疗慢性应激的有害影响。大学拟议的培训马里兰州巴尔的摩，医学院将有助于我过渡到博士后奖学金，并允许我通过表征神经元，小胶质细胞和在压力暴露的背景下，细胞外基质关系。在我出色的科学的支持下由我的导师，论文委员会成员，合作者和神经科学组成的社区以及 Meyerhoff社区，我很高兴完成F99/K00阶段中概述的技术和职业目标我的提议，工作并成长为独立研究者在神经科学中的作用。