Intranasal Treatment of Stem Cell-derived Extracellular Vesicles for Alzheimer's Disease

项目摘要 该项目的主要目标是开发一种非侵入性的，基于细胞外囊泡（EV）的治疗策略 用于改善阿尔茨海默病（AD）的大脑功能。一种新的方法，调查的有效性， 鼻内（IN）施用由人诱导多能干细胞（hiPSC）产生的治疗性EV- 提出了在AD小鼠模型中的衍生神经干细胞（NSC）。科学前提是hNSC- 携带有益的miRNA和神经保护蛋白的货物的衍生EV（hNSC-EV）可以激活 靶细胞中的有利信号传导途径，积极调节脑微环境，小胶质细胞， 和神经发生，并改善受伤或疾病后的脑功能。值得注意的是， 已经显示IN给予hNSC-EV导致它们被神经元和小胶质细胞掺入， 在5XFAD小鼠的几乎所有大脑区域，并导致更好的认知和情绪功能，更高水平的 海马神经发生，以及氧化应激、神经炎症和淀粉样蛋白沉积的减少。这 使用AD的小鼠模型，本项目将测试以下假设：IN施用hNSC衍生的EV：（i）在 AD早期将维持较好的认知和情绪功能;以及（ii）AD晚期将 逆转认知和情绪功能障碍，并显著调节神经病理学。特定目标研究 1将使用5XFAD小鼠并研究在AD的早期阶段用hNSC-EV干预是否会 保持更好的认知和情绪功能，以及这种积极影响是否会持续很长时间。 使用5XFAD和A-β-敲入小鼠的特定目标2的研究将测试IN是否 在AD的晚期施用hNSC-EV将逆转认知和情绪功能障碍。 具体目标3研究将确定小胶质细胞调节在hNSC-EV介导的改善中的作用。 通过使用PLX 5622引入的选择性小胶质细胞耗竭， 通过饮食。具体目标4中的研究将首先检查hNSC-EV的生物学特性的变化 使用hiPSC衍生的小胶质细胞培养物敲低或过表达特定的miRNA和蛋白质。 接下来，研究了IN施用过表达选择的miRNA和/或蛋白质的hNSC-EV的作用，所述选择的miRNA和/或蛋白质具有 将在6个月大的5XFAD小鼠中测试强的抗肿瘤活性，以确定这种抗肿瘤活性是否 该策略将改善hNSC-EV在AD晚期的治疗益处。男性和 使用雌性小鼠。海马体和内侧前额叶皮层将被严格检查 在所有目的中，由hNSC-EV介导的各种细胞和分子变化。特别 hNSC-EV治疗的功能改善是否包括抑制氧化应激 和神经炎症，海马神经发生的更高水平，以及淀粉样蛋白β沉积的减少， 将确定p-tau、突触丢失和神经变性。建议的翻译研究 高度有助于开发用于AD的同种异体hNSC-EV疗法。

Project Summary This project's principal goal is to develop a non-invasive, extracellular vesicle (EV) based therapeutic strategy for improving brain function in Alzheimer's disease (AD). A novel approach that investigates the efficacy of intranasal (IN) administration of therapeutic EVs generated from human induced pluripotent stem cell (hiPSC)- derived neural stem cells (NSCs) in mouse models of AD are proposed. The scientific premise is that hNSC- derived EVs (hNSC-EVs) carrying a cargo of beneficial miRNAs and neuroprotective proteins can activate advantageous signaling pathways in target cells, positively modulate the brain microenvironment, microglia, and neurogenesis in the brain, and improve brain function after injury or disease. Notably, preliminary studies have shown that IN administration of hNSC-EVs results in their incorporation by neurons and microglia in virtually all brain regions in 5XFAD mice and leads to better cognitive and mood function, higher levels of hippocampal neurogenesis, and reductions in oxidative stress, neuroinflammation, and amyloid deposits. This project, using mouse models of AD, will test the hypothesis that IN administration of hNSC derived EVs: (i) in the early stage of AD will maintain better cognitive and mood function; and (ii) in the advanced stage of AD will reverse cognitive and mood dysfunction with significant modulation of neuropathology. Studies in Specific Aim 1 will employ 5XFAD mice and investigate whether intervention with hNSC-EVs in the early stage of AD would maintain better cognitive and mood function and whether such positive effects persist for prolonged periods. Investigations in Specific Aim 2, using both 5XFAD and A-beta-Knock-in mice, will test whether IN administration of hNSC-EVs in the advanced stage of AD would reverse cognitive and mood dysfunction. Specific Aim 3 studies will ascertain the role of microglial modulation in hNSC-EV mediated improvements in cognitive and mood function in 5XFAD mice through selective microglia depletion using PLX5622 introduced through diet. Studies in Specific Aim 4 will first examine changes in the antiinflammatory property of hNSC-EVs with knock-down or overexpression of specific miRNAs and proteins, using hiPSC-derived microglia cultures. Next, the effects of IN administration of hNSC-EVs overexpressing the select miRNA and/or protein having robust antiinflammatory activity will be tested in 6 months old 5XFAD mice to determine whether such a strategy would improve the therapeutic benefits of hNSC-EVs in the advanced stage of AD. Both male and female mice will be employed. The hippocampus and the medial prefrontal cortex will be rigorously examined for the various cellular and molecular changes mediated by hNSC-EVs in all Aims. Particularly whether functional improvements with hNSC-EV treatment would comprise the suppression of oxidative stress and neuroinflammation, a higher level of hippocampal neurogenesis, and reductions in amyloid-beta deposits, p-tau, synapse loss, and neurodegeneration will be ascertained. The proposed translational research studies are highly conducive to developing an allogeneic hNSC-EV therapy for AD.