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

• 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
• 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

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.

项目概要 暴露于慢性压力是神经精神疾病的已知危险因素，并会产生结构和分子损伤 大脑和免疫系统的变化。伏隔核 (NAc)，整合奖赏和奖励的枢纽 动机，表现出在压力患者死后组织中发现的分子和细胞改变 相关疾病，例如重度抑郁症，会导致啮齿类动物的动机缺陷。具体来说， 表达中型多巴胺神经元的 1 型 (D1) 多巴胺受体 (D1) 树突状萎缩，但不包括 2 型 (D2) 多巴胺受体树突萎缩 （MSN）对于压力引起的负面行为结果是必要且充分的。新出现的证据 表明小胶质细胞是社会压力后神经元萎缩和动机缺陷的潜在调节因素。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 研究慢性压力和 D1-MSN 萎缩之间的关系。实验室最近的工作表明 慢性社交失败压力 (CSDS) 减少了 D1-MSN 和小胶质细胞的整体接触，但没有减少 D2 MSN，并且 对 D1-MSN 微环境中单个小胶质细胞的分析揭示了小胶质细胞的减少 CSDS 后表现出负面情感行为的动物的复杂性。因此，虽然 CSDS 显然 改变 D1-MSN-小胶质细胞接触和形态，驱动这些细胞亚型特异性的分子机制， 应激引起的 NAc 微环境变化仍不清楚。初步 RNA-seq 分析 实验室指出玻连蛋白（一种细胞外基质底物）作为介导 D1- 的有前途的分子信使 MSN 和小胶质细胞相互作用，因为它在慢性压力改变的基因网络中发挥驱动作用 并且在 D1-MSN 中特异性差异表达。在这个提案中，我将通过病毒方式敲除玻连蛋白 在小鼠中使用 Cre 依赖性 CRISPR/Cas9 构建体在 D1-MSN 中表达，然后再将它们置于 CSDS 或慢性见证失败压力 (CWDS) 来询问 D1-MSN- 中玻连蛋白表达的作用 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 假设敲低 D1-MSN 中玻连蛋白的表达将挽救 D1-MSN 应激诱导的 树突萎缩，恢复 D1-MSN-小胶质细胞接触，并防止负面情感行为。理解 驱动压力诱导行为状态改变的分子机制可以为新疗法带来启示 旨在保护和/或治疗慢性压力的有害影响。拟议的大学培训 马里兰州巴尔的摩医学院将促进我向博士后奖学金的过渡，并允许我 通过表征神经元、小胶质细胞和神经精神疾病，继续研究与压力相关的神经精神疾病 压力暴露背景下的细胞外基质关系。在我出色的科学知识的支持下 社区由我的导师、论文委员会成员、合作者以及神经科学和 Meyerhoff 社区，我很高兴能够完成 F99/K00 阶段中概述的技术和职业目标 我的建议以及工作并成长为神经科学领域独立研究者的角色。