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: 10582681
• 负责人: Srikant Rangaraju
• 金额: $ 41.99万
• 依托单位: EMORY 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 2023-10-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10582681
• 关键词: 3xTg-AD mouse; Acute; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease therapy; Amyloid beta-Protein; Anti-Inflammatory Agents; Apoptosis; Astrocytosis; Autoimmune Diseases; Award; Behavioral; Biological Assay; Biological Markers; Biological Response Modifiers; Biology; Bone Marrow; Brain; Cells; Central Nervous System; Cerebrospinal Fluid; Chimera organism; Coupled; Data; Dementia; Dependence; Disease; Disease Progression; Flow Cytometry; Foundations; Future; Gene Deletion; Goals; Health Expenditures; Hematopoietic; Human; Immune; Immune signaling; Immunophenotyping; Immunotherapeutic agent; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Kv1.3 potassium channel; Lead; LoxP-flanked allele; MAP Kinase Gene; Macrophage; Marrow; Mass Spectrum Analysis; Mediating; Microglia; Modeling; Mononuclear; Mus; Nerve Degeneration; Neurodegenerative Disorders; Neuroimmunomodulation; Outcome; PTPRC gene; Pathologic; Pathology; Pathway interactions; Peripheral; Phagocytes; Pharmacology; Phase; Phenotype; Pre-Clinical Model; Research Personnel; Role; Safety; Shapes; Signal Pathway; Stimulus; Synapses; Testing; Therapeutic; Therapeutic Effect; Therapeutic Studies; Translating; Translations; Treatment Efficacy; Wood material; abeta accumulation; analog; behavioral outcome; biomarker discovery; brain cell; cell type; conditioned fear; efficacy testing; experimental study; genetic approach; glial activation; immune function; immunoregulation; improved; in vivo; intervention effect; irradiation; monocyte; mouse model; multidisciplinary; neuro-immunomodulatory therapies; neurobehavioral; neuroimmunology; neuroinflammation; neuronal survival; neuropathology; new therapeutic target; novel; object recognition; p38 Mitogen Activated Protein Kinase; pharmacologic; pre-clinical; preclinical efficacy; protein biomarkers; radioresistant; ratiometric; reconstitution; release of sequestered calcium ion into cytoplasm; response; selective expression; small molecule; tau Proteins; tau aggregation; therapeutic target; transcriptomics; translation to humans

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（Alzheimer's disease，AD）是一种最常见的神经退行性疾病，可导致痴呆、依赖性痴呆、精神分裂症、认知障碍、认知障碍等 和医疗保健支出。大脑中的炎症（神经炎症）塑造了 通过促炎性（有害的）以及抗炎性（有害的）影响神经元存活， 炎症（保护）作用。中枢神经系统（CNS）单核吞噬细胞（CNS MP）是 由CNS驻留的小胶质细胞和CNS浸润的单核细胞组成），并且是 神经炎症在AD中选择性抑制促炎性CNS MP可能会减缓炎症反应的速率。 我们已经发现阻断钾通道Kv1.3是一种有前途的免疫策略 在AD中。Kv1.3通道调节钙流，在AD中由促炎性CNS MP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通道是 调节免疫信号传导和生存的促炎性中枢神经系统MP在AD中，可以治疗 选择性Kv1.3阻滞剂的靶向作用。在本提案中，我们将解决与角色相关的三个具体问题 AD中促炎性CNS MP表达的Kv1.3通道，总体目标是奠定临床前 为未来将Kv1.3阻滞剂转化为人类奠定了基础。首先，我们将进行体内实验， 在Aβ蓄积的5xFAD模型中使用Kv1.3阻断剂，以鉴定CNS MP中的免疫信号传导途径， 需要Kv1.3通道（目标1a）。我们还将确定阻断Kv1.3是否会降低促炎性细胞因子的表达。 通过增加细胞凋亡并将其转换为抗炎特性，在脑中产生CNS MP（目的1b）。 接下来，我们将检验表达大量Kv1.3通道的CNS MP来源于血液的假设 单核细胞，而不是小胶质细胞（目标2）。我们将对中枢神经系统MP和血液单核细胞进行转录组学研究， 并应用放射性骨髓嵌合体模型来确定AD模型中CNS MP是否起源于 从小胶质细胞或从血液单核细胞渗透到大脑。在目标3中，我们将进行长期体内研究 在两种AD小鼠模型中测试Kv1.3阻断剂对行为和病理终点的功效， 对脑脊液进行质谱分析，以鉴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，我们将使用一种新的方法来确定CNS MP Kv1.3通道在AD病理学中的作用。 AD模型中的条件缺失遗传方法。为达致R01建议的目的，我们 组建了一个多学科的研究团队，他们在神经免疫学、Kv1.3生物学和 神经变性小鼠模型（Rangaraju博士，PI），免疫信号传导（Wood博士），质谱和 生物标志物发现（Drs Seyfried和Levey）以及小分子药理学（Drs. Wulff）。