Targeting Kv1.3 potassium channels for neuro-immunomodulation in Alzheimer's Disease

靶向 Kv1.3 钾通道用于阿尔茨海默病的神经免疫调节

• 批准号: 10986277
• 负责人: Srikant Rangaraju
• 金额: $ 18.13万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10986277
• 关键词: Targeting; Kv1.3; potassium; channels; neuro

Alzheimer’s disease (AD) is the most common neurodegenerative disease resulting in dementia, dependence and healthcare expenditure. Inflammation in the brain (neuroinflammation) shapes the course of neurodegeneration by impacting neuronal survival through pro-inflammatory (detrimental) as well as anti- inflammatory (protective) effects. Central nervous system (CNS) mononuclear phagocytes (CNS MPs) are comprised of CNS-resident microglia and CNS-infiltrating monocytes), and are primary enactors of neuroinflammation. Selective inhibition of pro-inflammatory CNS MPs in AD may slow down the rate of neurodegeneration and we have found that blockade of potassium channel Kv1.3 is a promising immune strategy in AD. Kv1.3 channels regulate calcium flux, are expressed selectively by pro-inflammatory CNS MPs in AD and their blockade in AD mouse models limits neuropathology. We hypothesize that Kv1.3 channels are key regulators of immune signaling and survival in pro-inflammatory CNS MPs in AD that can be therapeutically targeted by selective Kv1.3 blockers. In this proposal, we will tackle three specific questions related to the role of Kv1.3 channels expressed by pro-inflammatory CNS MPs in AD, with the overall goal of laying the pre-clinical foundation for future translation of Kv1.3 blockers to humans. First, we will perform in-vivo experiments using Kv1.3 blockers in the 5xFAD model of Aβ accumulation to identify immune signaling pathways in CNS MPs that require Kv1.3 channels (Aim 1a). We will also determine whether blockade of Kv1.3 decreases pro-inflammatory CNS MPs in the brain by increasing apoptosis and switching them towards anti-inflammatory profiles (Aim 1b). Next, we will test the hypothesis that CNS MPs expressing high numbers of Kv1.3 channels originate from blood monocytes, rather than microglia (Aim 2). We will perform transcriptomics of CNS MPs and blood monocytes and apply the irradiation bone marrow chimera model to determine whether CNS MPs in AD models originate from microglia or from blood monocytes that infiltrate the brain. In Aim 3, we will perform long-term in-vivo studies to test the efficacy of Kv1.3 blockers on behavioral and pathological endpoints in two AD mouse models and will perform mass spectrometry of cerebrospinal fluid to identify protein biomarkers associated with therapeutic effects of Kv1.3 blockers. Lastly, we will determine the role of CNS MP Kv1.3 channels in AD pathology using a conditional deletion genetic approach in AD models. To achieve the aims of this R01 proposal, we have assembled a multi-disciplinary team of investigators with expertise in neuro-immunology, Kv1.3 biology and mouse models of neurodegeneration (Dr. Rangaraju, PI), immune signaling (Dr Wood), mass spectrometry and biomarker discovery (Drs Seyfried and Levey) as well as small-molecule pharmacology (Drs. Wulff).

阿尔茨海默病(AD)是一种最常见的导致痴呆、依赖的神经退行性疾病 和医疗保健支出。大脑中的炎症(神经炎症)塑造了 通过促炎(有害)和抗炎影响神经元存活的神经变性 发炎(保护)作用。中枢神经系统(CNS)单核巨噬细胞(CNS MPS) 由CNS驻留的小胶质细胞和CNS浸润性单核细胞组成)，是 神经炎。选择性抑制阿尔茨海默病中促炎症的中枢神经系统MPS可能减慢 我们发现阻断钾通道Kv1.3是一种很有前途的免疫策略 在公元后。Kv1.3通道调节钙流，在AD和AD中由促炎症的CNS MPS选择性表达 它们在AD小鼠模型中的阻断限制了神经病理学。我们假设Kv1.3渠道是关键 阿尔茨海默病中促炎症中枢神经系统MPS的免疫信号和生存调节可用于治疗 被选择性的Kv1.3阻滞剂瞄准。在这项提案中，我们将解决与角色相关的三个具体问题 在AD中由促炎症的CNS MPS表达的Kv1.3通道，总体目标是奠定临床前 为将来将Kv1.3阻滞剂翻译成人类奠定了基础。首先，我们将进行体内实验，使用 Kv1.3阻滞剂在Aβ蓄积的5xFAD模型中识别中枢神经系统MPS中的免疫信号通路 需要Kv1.3频道(目标1a)。我们还将确定封锁Kv1.3是否会减少促炎作用 中枢神经系统MPS通过增加细胞凋亡和将其转换为抗炎特征(目标1b)。 接下来，我们将检验一种假设，即高表达Kv1.3通道的CNS MPS来自血液 单核细胞，而不是小胶质细胞(目标2)。我们将进行中枢神经系统MPS和单核细胞的转录 并应用放射骨髓嵌合体模型确定AD模型中中枢神经系统MPS是否起源 来自小胶质细胞或渗入大脑的单核细胞。在目标3中，我们将进行长期的体内研究 测试Kv1.3阻滞剂对两种AD小鼠模型和Will的行为和病理终点的疗效 对脑脊液进行质谱分析以确定与治疗相关的蛋白质生物标志物 Kv1.3阻滞剂的作用。最后，我们将使用一种方法来确定CNS MP Kv1.3通道在AD病理中的作用 阿尔茨海默病模型中的条件缺失遗传方法。为达致R01建议的目标，我们已 组建了一支由神经免疫学、Kv1.3生物学和 神经变性的小鼠模型(Rangaraju博士，Pi)，免疫信号(Wood博士)，质谱学和 生物标记物的发现(塞弗里德博士和利维博士)以及小分子药理学(伍尔夫博士)。