Microglial and macrophage potassium channels as regulators of neuroinflammation in Alzheimer's Disease

项目摘要/摘要 侯选人 我是一名获得董事会认证的神经科医生，致力于面向健康的基础和转化性研究。我 在离子通道生物学和免疫学、临床研究方法和临床方面具有研究专长 神经学。我的长期目标是将自己确立为一名成功的独立内科医生兼科学家 神经炎症和转化性神经科学。为了实现这一目标，并促进我向 独立，我通过NIH K08职业发展奖寻求5年的指导培训，以填补空白 我的研究培训，建立一个未来研究合作者的网络，建立我的出版记录 提供助学金写作技能，并继续进行领导力和教师发展方面的培训。 环境 埃默里的培训和指导环境非常适合我的职业发展。我的导师之道 埃默里大学的一个委员会由以下领域的专家和独立科学家组成 神经退行性变(Allan Levey博士)、神经炎症(Malu Tansey博士)和蛋白质组学/系统生物学 (尼古拉斯·塞弗里德博士)，每个人都有NIH和非联邦资金的模范记录，领导着成功的 富有成效的研究小组和培训博士后和医生-科学家培训生。我的导师和他们的 研究小组将为我提供以下方面的实践培训：(1)神经退行性变动物模型作为工具 促进翻译研究，(2)小胶质细胞和巨噬细胞的功能分析，免疫信号， 免疫分析、神经胶质生物学，以及(3)蛋白质组学、生物信息学和系统生物学。我会接受研究 通过每周和每月的互动和对我的研究进展的直接监督来指导我， 在期刊俱乐部、研讨会上发表演讲，在地方和国家会议上发表演讲。我也会收到 神经胶质生物学、免疫学、神经科学研究的高级方法、生物信息学、 R编程、系统生物学(加权相关网络分析)和负责任的行为 埃默里大学(Emory University)的研究(12学分学时和满足NIH要求的面对面培训)。一支由 具有系统生物学、多肽合成、钾通道等方面专业知识的校内外人员 生物学和生物统计学也将指导拟议的研究。埃默里大学是一所大型学术机构 神经科学和神经病学获得了美国国立卫生研究院最高的研究经费之一(在美国排名第9)。这个 埃默里神经退行性疾病中心(CND)是一项合作和跨学科的研究 由美国国立卫生研究院资助的研究核心设施支持的共享实验室和研究资源的环境 在神经病理学、蛋白质组学、多重免疫分析、流式细胞术、细胞成像和啮齿动物 神经行为方法。 研究 阿尔茨海默病(AD)是导致痴呆症的最常见原因，目前缺乏疾病修正疗法 全世界。AD等疾病中单核巨噬细胞(MPS)介导的免疫应答 越来越多的人认识到，神经退行性疾病通过促进神经退行性变而影响神经退行性变。 发炎(促进疾病)和抗炎(保护)机制。前列环素选择性抑制剂 节省和/或促进抗炎作用的炎症功能可能会导致新的 治疗阿尔茨海默病和其他神经退行性疾病的疾病修正疗法。钾通道 MPS在大脑中的表达是免疫功能的关键调节因素，我们的初步数据表明 促炎症MP功能受Kv1.3钾通道调节，而抗炎MP功能受Kv1.3钾通道调节 功能受另一种名为Kir2.1的K通道调节。在这项提案中，我们将首先确定 Kv1.3和Kir2.1通道在促炎和抗炎MP功能中的功能重要性包括 体外产生活性氧，细胞因子/趋化因子的产生；并将研究 MPS中Kv1.3和Kir2.1通道阻断的流信号和转录效应(目标1)。接下来，我们将 测定外周血单核细胞来源的这两种钾通道的表达模式 从小鼠AD脑中分离出巨噬细胞和小胶质细胞，并将测试这些通道是否调节 在AD病理的小鼠模型中，外周来源的MPS进入大脑的能力(目标2)。最后，我们 将对Kv1.3和Kir2.1通道的慢性阻断进行初步研究，使用新型高选择性和 有效的阻滞剂作为治疗策略减轻5xFAD的神经退行性变和认知缺陷 阿尔茨海默病(AD)小鼠模型(目的3)。这项工作的结果将为小胶质细胞和巨噬细胞提供新的见解 神经退行性疾病的生物学、免疫调节和潜在的新治疗策略 这可以改变AD的疾病进展。从这些研究目标产生的数据也将推动 候选人的职业发展，并将导致未来的独立调查员(R01)资助和未来 在神经退行性疾病和神经炎症性疾病的范围内进行合作研究。

Project summary/Abstract Candidate I am a board-certified neurologist committed to a career in health-oriented basic and translational research. I have research expertise in ion channel biology and immunology, clinical research methods and clinical neurology. My long-term goal is to establish myself as a successful independent physician-scientist in neuroinflammation and translational neuroscience. To achieve this goal and facilitate my transition to independence, I seek 5 years of mentored training through a NIH K08 career development award to fill gaps in my research training, establish a network of future research collaborators, build my publication record and grant-writing skills and pursue training in leadership and faculty development. Environment The training and mentoring environment at Emory is ideally suited for my career development. My mentorship committee at Emory University is comprised of experts and independent scientists in the fields of neurodegeneration (Dr. Allan Levey), neuroinflammation (Dr. Malu Tansey) and proteomics/systems biology (Dr. Nicholas Seyfried), each with exemplary records of NIH and non-federal funding, leading successful and productive research groups and training post-doctoral and physician-scientist trainees. My mentors and their research groups will provide me with hands-on training in (1) Animal models of neurodegeneration as tools to facilitate translational research, (2) Functional assays of microglia and macrophages, immune signaling, immunoassays, glial biology, and (3) Proteomics, bioinformatics and systems biology. I will receive research mentorship through weekly and monthly interactions and direct oversight of my research progress, presentations at journal clubs, seminars and presentations at local and national meetings. I will also receive formal coursework in glial biology, immunology, advanced methods in neuroscience research, bioinformatics, R programing, systems biology (WeiGhted Correlation Network Analysis) and in the responsible conduct of research (>12 credit hours and in-person training that fulfils NIH requirements) at Emory University. A team of intra- and extra-mural contributors with expertise in systems biology, peptide synthesis, potassium channel biology and biostatistics will also guide the proposed research. Emory University is a large academic institution with Neuroscience and Neurology receiving one of highest research funding from the NIH (9th in the US). The Emory Center for Neurodegenerative Diseases (CND) is a collaborative and cross-disciplinary research environment with shared laboratory and research resources supported by NIH-funded research core facilities in neuropathology, proteomics, multiplexed immunoassays, flow cytometry, cell imaging and rodent neurobehavioral methods. Research Disease-modifying therapies are lacking for Alzheimer’s disease (AD), the most common cause of dementia worldwide. Immune responses mediated by mononuclear phagocytes (MPs) in AD and other neurodegenerative diseases are increasingly recognized to influence neurodegeneration through pro- inflammatory (disease-promoting) and anti-inflammatory (protective) mechanisms. Selective inhibitors of pro- inflammatory functions that spare and/or promote anti-inflammatory effects may lead to development of novel disease-modifying therapies for AD as well as other neurodegenerative diseases. Potassium channels expressed by MPs in the brain are key regulators of immune functions and our preliminary data suggest that pro-inflammatory MP functions are regulated by Kv1.3 potassium channels while anti-inflammatory MP functions are regulated by a different K channel called Kir2.1. In this proposal, we will first determine the functional importance of Kv1.3 and Kir2.1 channels in pro- and anti-inflammatory MP functions including reactive oxygen species production, cytokine/chemokine production in-vitro; and will investigate the down- stream signaling and transcriptional effects of Kv1.3 and Kir2.1 channel blockade in MPs (Aim 1). Next, we will determine expression patterns of these two potassium channels in peripheral blood monocyte-derived macrophages and microglia isolated from the mouse AD brain and will test whether these channels regulate the ability of peripherally derived MPs to enter the brain in a mouse model of AD pathology (Aim 2). Lastly, we will perform initial studies of chronic blockade of Kv1.3 and Kir2.1 channels using novel highly-selective and potent blockers as therapeutic strategies to mitigate neurodegeneration and cognitive deficits in the 5xFAD mouse model of AD (Aim 3). The results of this work will provide novel insights into microglial and macrophage biology, immune regulation in neurodegenerative diseases and potentially identify novel therapeutic strategies that can modify disease progression in AD. The data generated from these research aims will also advance the candidate’s career development and will lead to future independent investigator (R01) grants and future collaborative research across the spectrum of neurodegenerative and neuro-inflammatory diseases.