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

候选人：Jonathan Cherry博士的长期职业目标是成为一名独立的VA调查员， 建立一个研究实验室，重点研究头部创伤和神经退行性疾病。Cherry博士主要是 感兴趣的是在服兵役或接触性运动期间接受的重复性头部创伤如何有助于 神经退行性疾病慢性创伤性脑病（CTE）的发展和进展。 在拟议的CDA 2期间，Cherry博士将利用他的研究生和博士后经验， 神经炎症和炎症细胞，并扩大他的知识[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，以实现所提出的目标。在 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， 博士Cherry将每周接受脑解剖的实践培训。拟议的综合发展援助提供的支助2 将有助于塑造樱桃博士成为一个成功的VA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-脑震荡遗产基金会（VA-BU-CLF）大脑 VA波士顿的一个数据库包含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（Jamaica Plain VA hospital）的工作人员和高级科学家 波士顿）和波士顿大学创伤性脑病研究中心（BU CSTE）是世界上 CTE和神经变性方面的顶尖专家设施和人员提供了理想的环境， 执行本提案中所述的培训和研究。麦基博士是一位经验丰富的导师， 30多名各级研究人员后来取得了成功的职业生涯。许多中心附属于 VA波士顿和BU 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” 持久自由行动（Operation Enduring Freedom）此外，许多士兵将接受多个mTBI。重复性头部 创伤是发生CTE的最大风险因素，CTE是一种神经退行性疾病， 进行性记忆和行为障碍目前，CTE只能在死后诊断;因此， 需要了解哪些因素驱动病理学对于寻找诊断CTE的新生物标志物至关重要， 为活着的病人创造有效的治疗方法。神经炎症最近被牵连在CTE 病理进展;然而，机制仍不清楚。本研究提出的工作旨在 了解神经炎症如何可能导致CTE作为重复性头部创伤的结果。第一、 [Dr. Cherry将识别所有在重复性细胞移植后出现的炎症或神经退行性细胞群。 使用单核RNA测序（snRNA-seq）的mTBI（rmTBI）。Cherry博士将比较 使用来自没有rmTBI和神经退行性疾病史的个体的组织的神经炎性细胞群 有rmTBI病史但无神经退行性疾病的个体，以及有 rmTBI和CTE I或II期（即轻度CTE）。该分析将识别单个细胞群， 研究头部创伤后和早期CTE期间出现的重要机制细胞群体。这 这将是第一个使用有rmTBI病史的人脑进行snRNA-seq的研究。最后，Cherry博士将 使用多达9种颜色的多重染色，全面分析观察到的神经变性亚群 来可视化每个种群的区域位置。他将探索神经退行性亚群 与神经病理学特征如过度磷酸化tau、Aβ或TDP-43相互作用。 总的来说，本研究产生的结果对于促进对以下问题的基本理解是必要的： 头部创伤后神经炎症背后的机制，确定新的生物标志物来检测CTE，并设计 治疗活体疾病的方法。

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.