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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease model Alzheimer&apos s disease pathology Alzheimer&apos 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

项目摘要

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.

项目概要阿尔茨海默病（AD）是一种破坏性的神经退行性疾病，其特征是 β 淀粉样蛋白 (Aβ) 和 tau 蛋白、神经炎症、神经元损失和痴呆。小胶质细胞（大脑的免疫细胞）是治疗 AD 疾病的有希望的细胞靶标。用于治疗开发以改变小胶质细胞向独特状态转变的过程。 AD 中被称为疾病相关小胶质细胞 (DAM) 的疾病相关小胶质细胞通过保护性和 DAM 的一个子集（称为促炎性 DAM）会促进神经元损伤，并且无法发挥作用。初步研究表明促炎 DAM 发生生物能转变。因此，确定糖酵解状态以维持其应激反应。糖酵解转变对于确定抑制 AD 促炎症反应的新途径至关重要。发现促炎 DAM 高度表达称为 Kv1.3 的钾通道，该通道可调节钾我们实验室的工作表明 AD 小鼠模型中 Kv1.3 通道被阻断。病理学减少炎症和 Aβ 病理学，但 Kv1.3 如何调节促炎性 DAM 中的反应仍不清楚。我认为 Kv1.3 通过控制炎症反应来调节炎症反应。我的中心假设是 Kv1.3 通道是小胶质细胞的关键调节因子。我将在体外和体内使用小胶质细胞的生物能开关和促炎症反应。研究 Kv1.3 如何调节 DAM 的生物能学和炎症反应的方法。该项目的长期目标是研究 Kv1.3 在小胶质细胞生物能学和炎症中的作用。目标 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目标 1 将使用海马检测、活细胞成像、免疫来激活小胶质细胞的功能。分析和流式细胞术来描述生物能学和炎症反应的这些变化。我将利用一种新的遗传方法选择性地删除 AD 小鼠模型体内小胶质细胞中的 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的影响通过免疫分析、蛋白质印迹、关键代谢物检测来检测炎症反应，以及这两个平行的目标将使我能够阐明 Kv1.3 的机制。调节 AD 中的促炎性 DAM 反应通过了解小胶质细胞 Kv1.3 对 AD 的影响，我将有助于了解 AD 背后的生物学并建立潜在的治疗标记。我将在这个建议中获得培训，我将为我未来成为独立的职业目标做好充分的准备研究者。