Identification of the neuroinflammatory signature for CTE using single nucleus RNA sequencing

候选人：乔纳森·切里博士的长期职业目标是成为一名独立的退伍军人管理局调查员和 建立以头部创伤和神经退行性疾病为重点的研究实验室。切瑞博士主要是 感兴趣的是在服兵役或接触运动期间反复受到的头部创伤如何有助于 神经退行性疾病慢性创伤性脑病的发生发展。 在提议的CDA2期间，Cherry博士将建立他的研究生和博士后研究经验 神经炎症和炎性细胞，并扩大他的[生物信息学，单细胞RNA- 测序]、人类神经解剖学和临床疾病介绍，以实现拟议的目标。在……里面 除了正式的课程工作和与Ann McKee博士(导师)及其咨询委员会的定期会议外， 切里博士将每周接受脑部解剖方面的实践培训。拟议的CDA2提供的支持 将有助于将Cherry博士塑造成一名成功的退伍军人管理局调查员，并导致关键和及时的 关于退伍军人CTE的知识。 环境：退伍军人事务部-波士顿大学-震荡遗产基金会(VA-BU-CLF)大脑 弗吉尼亚州波士顿银行拥有世界上最大的神经病理学诊断的CTE病例队列， 代表了CTE研究的前沿。牙买加平原退伍军人医院(VA)的人员和高级科学家 波士顿大学创伤性脑病研究中心(BU CSTE)是世界上 CTE和神经退行性变领域的顶尖专家。设施和人员提供了理想的环境 执行本建议书中所述的培训和研究。McKee博士是一位经验丰富的导师，经过培训 30多名各个层次的研究人员，他们后来都有成功的职业生涯。附属的许多中心 退伍军人事务部波士顿分校和波士顿大学CSTE也将允许多种合作。 研究：轻度创伤性脑损伤(MTBI)被称为伊拉克行动的“标志性损伤” 自由/持久自由行动。此外，许多士兵将收到多个MTBI。重复磁头 创伤是发生CTE的最大风险因素，CTE是一种神经退行性疾病，其特征是 渐进性记忆和行为障碍。目前，CTE只能在尸检后诊断；因此， 需要了解推动病理的因素对于找到新的生物标记物来诊断CTE和 为活着的病人创造有效的治疗方法。神经炎症最近被认为与CTE有关 病理进展；然而，其机制仍不清楚。这项研究中提出的工作旨在 了解神经炎如何潜在地导致反复头部创伤导致的CTE。第一, [Cherry博士将确定重复复制后出现的所有炎性或神经变性细胞群 MTBI(RmTBI)使用单核RNA测序(SnRNA-seq)。切里博士将会比较 使用无rmTBI病史和神经退行性变患者组织的神经炎性细胞群 疾病，有rmTBI病史但无神经退行性疾病的个人，以及有 RmTBI和CTE I或II期(即轻度CTE)。这项分析将确定单个细胞群体和 研究头部创伤后和早期CTE期间出现的重要机械性细胞群。这 将是第一个使用有rmTBI历史的人脑进行SnRNA-SEQ的研究。最后，切瑞博士将 用多达9种颜色的多重染色综合分析观察到的神经退行性变亚群 以可视化每个人口的区域位置。他将探索神经退行性亚群是否 与神经病理特征相互作用，如过度磷酸化的tau、Aβ或Tdp-43。] 总体而言，这项研究产生的结果对于促进对 脑外伤后神经炎症背后的机制，寻找检测CTE的新生物标记物，并设计 治疗活体受试者疾病的疗法。

Candidate: The long-term career goal of Dr. Jonathan Cherry is to become an independent VA investigator and establish a research laboratory focused on head trauma and neurodegenerative diseases. Dr. Cherry is primarily interested in how repetitive head trauma received during military service or contact sports contributes toward the development and progression of the neurodegenerative disease chronic traumatic encephalopathy (CTE). During the proposed CDA2, Dr. Cherry will build off his graduate and postdoctoral experience studying neuroinflammation and inflammatory cells, and expand his knowledge of [bioinformatics, single cell RNA- sequencing], human neuroanatomy and clinical disease presentation to accomplish the proposed aims. In addition to formal course work and regular meetings with Dr. Ann McKee (Mentor) and his advisory committee, Dr. Cherry will receive weekly hands-on training in brain dissection. The support provided by the proposed CDA2 will be instrumental in shaping Dr. Cherry into a successful VA investigator, and result in critical and timely knowledge regarding CTE in Veterans. Environment: The Veterans Affairs – Boston University – Concussion Legacy Foundation (VA-BU-CLF) brain bank at VA Boston contains the world’s largest neuropathologically diagnosed cohort of CTE cases and represents the forefront of CTE research. Personnel and senior scientists at the Jamaica Plain VA hospital (VA Boston) and the Boston University Center for the Study of Traumatic Encephalopathy (BU CSTE) are the world’s leading experts in CTE and neurodegeneration. The facilities and personnel provide the ideal environment to perform the training and research described in this proposal. Dr. McKee is an experienced mentor, having trained over 30 researchers of all levels that have had subsequent successful careers. The many centers affiliated with VA Boston and the BU CSTE will also allow for multiple collaborations. Research: Mild traumatic brain injury (mTBI) has been called the “signature injury” of Operation Iraqi Freedom/Operation Enduring Freedom. Furthermore, many soldiers will receive multiple mTBIs. Repetitive head trauma is the single greatest risk factor for developing CTE, a neurodegenerative disease characterized by progressive memory and behavior impairments. Currently, CTE can only be diagnosed post-mortem; thus, the need to understand what factors drive pathology are critical to finding novel biomarkers to diagnose CTE and create effective therapeutics for living patients. Neuroinflammation has recently been implicated in CTE pathologic progression; however, the mechanisms are still unclear. The work proposed in this study aims to understand how neuroinflammation potentially leads to CTE as a consequence of repetitive head trauma. First, [Dr. Cherry will identify all the inflammatory or neurodegenerative cell populations that emerge after repetitive mTBI (rmTBI) using single nucleus RNA sequencing (snRNA-seq). Dr. Cherry will compare the neuroinflammatory cell populations using tissue from individuals with no history of rmTBI and neurodegenerative disease, individuals with history of rmTBI but no neurodegenerative disease, and individuals with a history of rmTBI and CTE stage I or II (i.e. mild CTE). This analysis will identify individual populations of cells and investigate important mechanistic cellular populations that emerge after head trauma and during early CTE. This will be the first study to perform snRNA-seq using human brains with a history of rmTBI. Finally, Dr. Cherry will comprehensively analyze the observed neurodegenerative subpopulations using up to 9 color multiplex staining to visualize the regional location of each population. He will explore if the neurodegenerative subpopulations have interactions with neuropathologic features such as hyperphosphorylated tau, Aβ, or TDP-43.] Overall, results generated from this study are necessary to progress the fundamental understanding of mechanisms behind neuroinflammation after head trauma, identify novel biomarkers to detect CTE, and design therapies to treat disease in living subjects.