Precision Identification and Targeting of Rod Microglia in Diffuse Brain-Injured Cortex

弥漫性脑损伤皮层中杆状小胶质细胞的精确识别和靶向

• 批准号: 10199762
• 负责人: Katherine R Giordano
• 金额: $ 4.01万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10199762
• 关键词: Adult; Alzheimer' s Disease; Animals; Anti-Inflammatory Agents; Antibodies; Antigens; Bacterial Infections; Biological Markers; Blood; Brain; Brain Injuries; Brain region; Cell surface; Cells; Chemical Exposure; Data; Detection; Diagnosis; Diffuse; Diffuse Brain Injury; Disease; Disease Progression; Disease model; Event; Evolution; Experimental Models; Fab Immunoglobulins; Female; Fluorescent in Situ Hybridization; Functional disorder; Gene Expression; Genes; Genomics; Harvest; Head; Histologic; Human; Immunohistochemistry; Immunoprecipitation; Individual; Inflammation; Inflammatory; Investigation; Ischemia; Laboratories; Mentors; Microglia; Microscopic; Modeling; Molecular; Molecular Profiling; Morphology; Mus; Nervous System Trauma; Neurodegenerative Disorders; Neurologic; Neurologic Symptoms; Neurons; Outcome; Paresis; Pathologic; Pathology; Pathway Analysis; Phage Display; Pharmacology; Population; Positioning Attribute; Process; Prognosis; Protocols documentation; Recovery; Reporting; Resolution; Rod; Rodent; Role; Seizures; Signal Transduction; Structure; Syphilis; Testing; Time; Training; Traumatic Brain Injury; Typhoid Fever; Variant; Visualization; Western Blotting; autism spectrum disorder; base; brain cell; brain repair; cytokine; experience; fluid percussion injury; injury and repair; injury recovery; laser capture microdissection; male; mechanical force; molecular marker; neuroinflammation; neurological recovery; neuronal cell body; neuropathology; next generation; next generation sequencing; repaired; tool; transcriptome sequencing

PROJECT SUMMARY: After experimental diffuse traumatic brain injury (TBI), we rediscovered the rod microglia – a morphological variant of activated microglia in the pathology or repair of the brain. Rod microglia are depicted in the drawings of Nissl, Cajal, and del Rio Hortega in cases of general paresis, but were largely ignored for around 100 years. Rod microglia have recently been visualized in human Alzheimer’s disease, Autism spectrum disorder and experimental models of TBI, bacterial infection, and seizure, but no function has been reported. We reproducibly observe rod microglia that align in trains across cortical layers adjacent to neuronal processes in foci of neuropathology after diffuse TBI and they form and recede in a time course consistent with neuropathology. Yet, advanced molecular tools are absent from the arsenal necessary to understand rod microglia structure and function. We hypothesize that rod microglia have a unique molecular profile compared to other microglia morphologies, which provide tools to confirm rod microglia mechanisms in diffuse TBI. To investigate the significance and impact of rod microglia in diffuse TBI, the current proposal will provide critical data to distinguish rod microglia from activated and ramified microglia by molecular profile using phage display biopanning and RNA sequencing. We will induce diffuse TBI by midline fluid percussion injury in adult male and female CX3Cr1-eGFP mice and identify rod microglia based on morphology. Laser capture microdissection will isolate rod microglia and non-rod microglia (activated and ramified) from cortical regions. Isolated cells in aim 1 will be used for phage display biopanning to develop and validate a cell surface marker unique to rod microglia. Isolated cells in aim 2 will be used for next-generation sequencing (RNAseq) to reveal upregulated gene expression in rod microglia compared to activated and ramified morphologies. Pathway analysis will be incorporated to identify unique expression domains associated with the function and origin of rod microglia. The sponsor and mentoring team provide collective experience and expertise to provide advanced training for the applicant to achieve the aims. Data from this proposal will define the molecular profile of rod microglia and result in developed tools for quantitative and isolation protocols of rod microglia. With refined molecular tools, rod microglia can be investigated across neurological conditions (e.g. Alzheimer’s disease, Autism spectrum disorder, seizure) and activated or inhibited to drive treatment of neurological symptoms.

项目概要： 在实验性弥漫性创伤性脑损伤（TBI）后，我们重新发现了视杆细胞小胶质细胞-一种形态学改变。 在大脑的病理或修复中激活的小胶质细胞的变体。图中描绘了杆状小胶质细胞 Nissl，Cajal和德尔里奥霍特加在一般轻瘫的情况下，但在很大程度上被忽视了约100年。 最近，在人类阿尔茨海默病、自闭症谱系障碍和阿尔茨海默病中发现了杆状小胶质细胞。 TBI，细菌感染和癫痫发作的实验模型，但没有功能的报道。 我们可重复地观察到杆状小胶质细胞在邻近神经元突起的皮质层中排成一列 在弥漫性TBI后的神经病理学病灶中，它们的形成和消退时间与 神经病理学然而，先进的分子工具是缺乏必要的武器库，以了解杆 小胶质细胞的结构和功能。我们假设杆状小胶质细胞具有独特的分子特征， 其他小胶质细胞形态，这提供了工具，以确认杆小胶质细胞机制弥漫性TBI。 为了研究视杆细胞小胶质细胞在弥漫性TBI中的意义和影响，目前的建议将提供关键的 利用噬菌体展示通过分子谱区分视杆细胞小胶质细胞与活化的和分支的小胶质细胞的数据 生物淘选和RNA测序。我们将在成年男性中通过中线液压冲击损伤诱导弥漫性TBI， 雌性CX 3Cr 1-eGFP小鼠，并基于形态学鉴定视杆细胞小胶质细胞。激光捕获显微切割将 从皮层区域分离杆状小胶质细胞和非杆状小胶质细胞（活化和分支）。aim 1中的分离细胞 将用于噬菌体展示生物淘选，以开发和验证杆状小胶质细胞特有的细胞表面标记。 目的2中分离的细胞将用于下一代测序（RNAseq），以揭示上调的基因 与活化的和分枝的形态相比，在视杆小胶质细胞中的表达。路径分析将 掺入以鉴定与视杆小胶质细胞的功能和起源相关的独特表达结构域。的 赞助商和指导团队提供集体经验和专业知识， 申请人实现目标。来自该提议的数据将定义视杆细胞小胶质细胞的分子概况， 在开发工具的定量和分离协议杆小胶质细胞。借助精密的分子工具， 小胶质细胞可以在神经系统疾病（例如阿尔茨海默病、自闭症谱系）中进行研究 病症、癫痫发作）并被激活或抑制以驱动神经症状的治疗。