Regulation of Microglial Bioenergetics and Neuroinflammation by Kv1.3 Channels in Alzheimer's Disease

阿尔茨海默病中 Kv1.3 通道对小胶质细胞生物能和神经炎症的调节

• 批准号: 10670430
• 负责人: Christine Alyssia Bowen
• 金额: $ 4.77万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10670430
• 关键词: Address; Adopted; Affect; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease therapy; American; Amyloid beta-Protein; Autopsy; Biochemical; Bioenergetics; Biological Assay; Biology; Brain; Cell Respiration; Cell physiology; Cells; Cellular Membrane; Cre-LoxP; Dementia; Deposition; Disease; Disease Progression; Disease associated microglia; Flow Cytometry; Future; Gene Expression; Genes; Genetic; Genus Hippocampus; Glycolysis; Goals; Human; Immune; Immune response; Immunohistochemistry; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Laboratory Finding; Link; LoxP-flanked allele; Measures; Mediating; Membrane Potentials; Metabolic; Microglia; Mitochondria; Molecular; Monitor; Morphology; Mus; Neurodegenerative Disorders; Neuronal Injury; Neurons; Oxidative Phosphorylation; Pathogenesis; Pathologic; Phagocytes; Pharmacology Study; Phenotype; Play; Potassium; Potassium Channel; Quantitative Reverse Transcriptase PCR; Regulation; Research; Research Personnel; Role; Small Interfering RNA; Synapses; System; Testing; Therapeutic; Training; Visualization; Western Blotting; Work; abeta accumulation; beta amyloid pathology; brain cell; career; cell type; cellular targeting; cytokine; experimental study; extracellular; genetic approach; immune function; in vivo; inhibitor; knock-down; live cell imaging; mitochondrial membrane; molecular phenotype; mouse model; neuroinflammation; neuron loss; novel; overexpression; pharmacologic; preservation; programs; protein aggregation; release of sequestered calcium ion into cytoplasm; response; tau Proteins; therapeutic biomarker; therapeutic development; therapeutic target

PROJECT SUMMARY Alzheimer’s disease (AD) is a devastating neurodegenerative disease is characterized by accumulation of amyloid-β (Aβ) and tau proteins, neuroinflammation, neuronal loss, and dementia. Currently, there are no disease-altering therapies for AD. Microglia, the immune cells of the brain, represent promising cellular targets for therapeutic development to modify the course of AD progression. Microglia transition to a unique state called disease associated microglia (DAM) in AD which contribute to AD pathogenesis via both protective and detrimental responses. A subset of DAM, known as proinflammatory DAM, promote neuronal injury and fail to phagocytize Aβ effectively. Preliminary studies indicate proinflammatory DAM undergo a bioenergetic shift toward a glycolytic state to sustain their detrimental responses. Therefore, identifying the regulators of the glycolytic shift is critical in determining novel avenues to inhibit proinflammatory responses in AD. Our lab found proinflammatory DAM highly express a potassium channel called Kv1.3, which regulates potassium efflux and cellular functions. Our lab’s work showed blockade of Kv1.3 channels in mouse models of AD pathology reduces neuroinflammation and Aβ pathology, but how Kv1.3 regulates the proinflammatory response in DAM remains unclear. I propose that Kv1.3 regulates inflammatory responses by controlling the bioenergetics of microglia. My central hypothesis is the Kv1.3 channel is a critical regulator of the bioenergetic switch and proinflammatory responses of microglia in AD pathology. I will use in vitro and in vivo approaches to examine how Kv1.3 modulates the bioenergetics and inflammatory responses of DAM. The long-term goal of this project is to examine the role of Kv1.3 in microglial bioenergetics and inflammation. In Aim 1, I will use an in vitro system to examine how Kv1.3 alters the microglial bioenergetics and immune function of microglia which will be activated by Aβ. Aim 1 will use seahorse assays, live cell imaging, immune profiling, and flow cytometry to describe these changes in bioenergetics and inflammatory response. In Aim 2, I will utilize a novel genetic approach to delete Kv1.3 selectively in microglia in vivo in an AD mouse model. This will allow me to measure the effects of Kv1.3 deletion in microglia on microglial bioenergetics and inflammatory responses via immune profiling, western blots, assays for key metabolites, and immunohistochemistry. These two parallel aims will allow me to elucidate a mechanism by which Kv1.3 regulates proinflammatory DAM responses in AD. By understanding Kv1.3 influence of microglia in AD, I will contribute to the understanding of biology behind AD and establish a potential therapeutic marker. Through the training I will gain in this proposal, I will be well prepared for my future career goal of becoming an independent investigator.

项目摘要 阿尔茨海默氏病（AD）是一种毁灭性神经退行性疾病的特征是 淀粉样蛋白（Aβ）和tau蛋白，神经炎症，神经元丧失和痴呆。目前，没有 AD改变疾病的疗法。小胶质细胞，大脑的免疫细胞，代表有望的细胞靶标 为了进行热发育来修改AD进程的过程。小胶质细胞过渡到独特的状态 AD中称为疾病相关的小胶质细胞（DAM），这些小胶质细胞（DAM）通过受保护和 有害的反应。大坝的一部分，称为促炎性大坝，促进神经元损伤，无法 有效地吞噬Aβ。初步研究表明经历了生物能量的促炎性大坝 走向糖酵解状态，以维持其有害的反应。因此，确定 糖酵解转移对于确定新型途径抑制AD中的促炎反应至关重要。我们的实验室 发现促炎大坝高度表达称为KV1.3的钾通道，该通道调节钾 外排和细胞功能。我们的实验室的工作显示了AD鼠标模型中KV1.3通道的封锁 病理学降低了神经炎症和Aβ病理学，但是KV1.3如何调节促炎性 大坝中的反应尚不清楚。我建议KV1.3通过控制 小胶质细胞的生物能学。我的中心假设是KV1.3通道是 小胶质细胞在AD病理学中的生物能开关和促炎反应。我将在体外和体内使用 检验KV1.3如何调节大坝的生物能和炎症反应的方法。这 该项目的长期目标是检查KV1.3在小胶质细胞生物能学和炎症中的作用。在 AIM 1，我将使用体外系统来检查KV1.3如何改变小胶质细胞生物能和免疫 小胶质细胞的功能，该功能将被Aβ激活。 AIM 1将使用Seahorse测定，实时细胞成像，免疫 分析和流式细胞仪描述生物能和炎症反应中的这些变化。在AIM 2中， 在AD小鼠模型中，我将利用一种新型的遗传方法在体内选择性地删除KV1.3。 这将使我能够测量小胶质细胞中KV1.3缺失对小胶质细胞生物能和 通过免疫分析，蛋白质印迹，关键代谢产物的测定法和炎症反应以及 免疫组织化学。这两个平行目的将使我能够阐明一种机制KV1.3 调节AD中的促炎大坝反应。通过了解KV1.3小胶质细胞在AD中的影响，我将 有助于理解AD背后的生物学并建立潜在的治疗标记。通过 我将在此提案中获得培训，我将为成为独立的职业目标做好充分的准备 研究者。