Crosstalk between innate-immunity human microglia and adaptive-immunity Tregs in Alzheimer's disease

阿尔茨海默病中先天免疫人类小胶质细胞和适应性免疫 Tregs 之间的串扰

• 批准号: 10515987
• 负责人: STUART A LIPTON
• 金额: $ 44.38万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10515987
• 关键词: Adaptive Immune System; Affect; Alzheimer' s Disease; Alzheimer' s disease brain; Amyloid beta-Protein; Anti-Inflammatory Agents; Astrocytes; Binding; Blood; Blood donor; Bone Marrow; Brain; Cell Death; Cells; Cessation of life; Coculture Techniques; Cues; Development; Disease; Electrophysiology (science); Excitatory Amino Acid Antagonists; Exposure to; Fluorescent Probes; Functional disorder; Future; Gene Expression; Glutamate Receptor; Glutamates; Human; Immune; Immune response; Inflammation; Inflammatory; Inflammatory Response; Interleukin-10; Ion Channel; Knowledge; Lead; Light; Measures; Memantine; Methods; Microglia; Mutation; N-Methylaspartate; NMDA receptor A1; Natural Immunity; Neuraxis; Neurodegenerative Disorders; Neurons; Pathogenesis; Pathway interactions; Persons; Prevalence; Protocols documentation; Publishing; Receptor Activation; Regulatory T-Lymphocyte; Reporting; Signal Transduction; Stimulus; Synapses; System; T-Lymphocyte; Testing; Time; Transforming Growth Factor beta; Western Blotting; Work; Yolk Sac; acquired immunity; adaptive immunity; alpha synuclein; base; cell motility; chemokine; cytokine; exposed human population; gene correction; human old age (65+); induced pluripotent stem cell; insight; macrophage; migration; misfolded protein; neuroinflammation; neuronal survival; novel therapeutic intervention; patch clamp; prevent; receptor expression; recruit; response

Title: Crosstalk between innate-immunity human microglia and adaptive-immunity Tregs in Alzheimer’s disease (RFA-AG-22-017) Project Summary This application is in response to RFA-AG-22-017, looking at crosstalk between innate and acquired immunity in Alzheimer’s disease (AD). AD affects ~5% of people over age 65, and its prevalence is increasing. Neuroinflammation has been described in neurodegenerative disorders, and particularly in AD. Microglial cells, the resident macrophages of the central nervous system (CNS), respond to different pro- inflammatory stimuli by releasing glutamate, as well as pro-inflammatory cytokines and chemokines. Glutamate release from astrocytes and microglia has also been observed after exposure to Aβ and to αSyn, proteins that are misfolded in AD. T lymphocytes, express several NMDA-type glutamate receptors (NMDARs), and we have also detected their expression in Tregs, whose activity can reportedly ameliorate neurodegenerative disorders. Using patch-clamp recording, our Preliminary Studies show for the first time the presence of functional NMDAR-operated ion channels on human Tregs. Glutamate is known to affect T cell migration and activation; however, the effect of glutamate on Treg migration/activation has not been reported, and, specifically, it is not known how this might impact neurons in AD brain. We hypothesize that glutamate exposure of Tregs will limit their release of anti-inflammatory and neuroprotective factors, thus contributing to neuronal synaptic damage and cell loss. This crosstalk between the innate and acquired immune system may represent a significant contributor to the pathogenesis of AD. We have recently developed a protocol for generating human iPSC-derived microglia (hiMG) following a yolk sac differentiation method (brain microglia originate from the yolk sac and not from the bone marrow). We will test our hypothesis using a platform of hiPSC-derived microglia (hiMG), human Tregs (isolated from normal blood donors), and hiPSC-derived cerebrocortical neurons (hiN) bearing AD mutations (vs. their isogenic/gene- corrected controls). Using this platform, we show in our Preliminary Studies that hiMG release glutamate following activation with oligomeric Aβ and αSyn. We will systematically investigate the mechanism of the activation of Tregs by glutamate released by hiMG, and the effect of this cellular crosstalk on neuronal function. We will evaluate neuronal synaptic integrity and function as well as neuronal survival. The knowledge gained from this study will shed light on the effect of glutamate on neuroinflammation in a human context, and will provide insight into the effect of crosstalk between human microglial and Tregs in stimulating Treg activation. Thus, the work may aid in the development of future novel therapeutic interventions based on human microglial and Treg function and interaction.

标题：阿尔茨海默病中先天免疫人类小胶质细胞和适应性免疫 Tregs 之间的串扰 疾病 (RFA-AG-22-017) 项目概要 此应用程序是为了响应 RFA-AG-22-017，着眼于先天和后天之间的串扰 阿尔茨海默病（AD）的免疫力。 AD 影响约 5% 的 65 岁以上人群，其患病率是 增加。神经炎症已被描述为神经退行性疾病，特别是 AD。 小胶质细胞是中枢神经系统 (CNS) 的常驻巨噬细胞，对不同的亲 通过释放谷氨酸以及促炎细胞因子和趋化因子来刺激炎症。谷氨酸盐 在暴露于 Aβ 和 αSyn 后，也观察到星形胶质细胞和小胶质细胞的释放，这些蛋白质 AD 中折叠错误。 T 淋巴细胞表达多种 NMDA 型谷氨酸受体 (NMDAR)，我们 还检测到它们在 Tregs 中的表达，据报道，Tregs 的活性可以改善神经退行性疾病 失调。使用膜片钳记录，我们的初步研究首次表明存在 人类 Tregs 上功能性 NMDAR 操作的离子通道。已知谷氨酸会影响 T 细胞迁移 激活;然而，谷氨酸对 Treg 迁移/激活的影响尚未有报道，并且， 具体来说，尚不清楚这会如何影响 AD 大脑中的神经元。 我们假设 Tregs 的谷氨酸暴露将限制它们释放抗炎和 神经保护因子，从而导致神经元突触损伤和细胞损失。这种串扰之间 先天性和获得性免疫系统可能是 AD 发病机制的重要贡献者。我们 最近开发了一种在卵黄囊后生成人 iPSC 衍生小胶质细胞 (hiMG) 的方案 分化方法（脑小胶质细胞起源于卵黄囊而不是骨髓）。我们将测试 我们的假设使用 hiPSC 衍生的小胶质细胞 (hiMG)、人类 Tregs（从正常血液中分离）的平台 供体）和携带 AD 突变的 hiPSC 衍生的脑皮质神经元（hiN）（与其同基因/基因相比） 修正控制）。使用这个平台，我们在初步研究中表明 hiMG 释放谷氨酸 用寡聚 Aβ 和 αSyn 激活后。我们将系统地研究其机制 hiMG 释放的谷氨酸激活 Tregs，以及这种细胞串扰对神经元的影响 功能。我们将评估神经元突触的完整性和功能以及神经元的存活率。 从这项研究中获得的知识将揭示谷氨酸对神经炎症的影响 人类背景，并将深入了解人类小胶质细胞和 Tregs 之间的串扰效应 刺激 Treg 激活。因此，这项工作可能有助于开发未来新型治疗干预措施 基于人类小胶质细胞和 Treg 的功能和相互作用。