Targeting CNS neuroinflammation in animal models of Alzheimer's disease

项目摘要/摘要 这是一份修改后的申请表。神经炎症已被确定为导致 阿尔茨海默病(AD)的发病机制。研究显示β斑块周围有激活的小胶质细胞 死后阿尔茨海默病的大脑提示炎症通路在疾病进展中有重要参与。宠物 用放射性标记的小胶质细胞激活示踪剂对AD患者进行成像证实了这些发现。 在用于研究AD的动物模型中，也观察到小胶质细胞参与的神经炎症。 然而，小胶质细胞在阿尔茨海默病中的研究因缺乏特异性分子而受到阻碍。 了解小胶质细胞的表型和功能，同样重要的是缺乏治疗方法 可以针对小胶质细胞和神经炎症的方法。我们假设神经炎是一种 阿尔茨海默病及其抑制小胶质细胞治疗的重要靶点 通过我们的新方法(鼻腔抗CD3)，我们将能够治疗阿尔茨海默病。 我们发现，鼻腔抗CD3抗体可诱导抗炎免疫反应，从而减弱 驻留在脑内的小胶质细胞中的神经炎症。在新的初步数据中，我们发现鼻腔抗CD3 改善了AD小鼠模型的短期记忆。我们发现鼻腔注射抗CD3抗体 定位于颈部淋巴结，在那里诱导分泌IL-10的T细胞，然后迁移到大脑和 抑制小胶质细胞的神经炎症。因此，鼻腔给药抗CD3是一种独特而未被探索的方法。 治疗阿尔茨海默病神经炎症的靶点。这项建议的目的是为了获得调查的潜力 鼻腔抗CD3抑制小胶质细胞神经炎症作为AD的治疗策略。 目的1.探讨鼻腔抗CD3抗体对小胶质细胞表型和吞噬功能的影响 衰老。给24月龄小鼠鼻腔注射抗CD3抗体，以评估治疗对体内小胶质细胞的影响 吞噬功能与小胶质细胞内稳态的恢复。我们还将测量IL-10的诱导- 在这些小鼠的外周分泌T细胞，这将成为一个容易测量的生物标记物，并在 IL-10在鼻腔抗CD3对衰老小胶质细胞功能影响中的作用此外，我们将使用 不表达小胶质细胞上IL-10受体的条件基因敲除小鼠 研究IL-10在小胶质细胞吞噬特性中的作用以及其降解抗体斑块的能力。 目的2.观察鼻腔抗CD3抗体对3xTg AD模型的影响。我们将执行详细的行为 鼻腔抗CD3治疗3xTg AD小鼠的分析及神经病理学研究 抗CD3抗体的治疗潜力。此外，我们还将研究鼻腔抗CD3治疗对慢性粒细胞白血病的影响。 利用高通量RNA测序(Smart-seq2)研究AD小鼠模型中的小胶质神经炎。

PROJECT SUMMARY/ABSTRACT This is a revised application. Neuroinflammation has been identified as a major contributing factor to the pathogenesis of Alzheimer’s disease (AD). Studies showing activated microglia surrounding Aβ plaques in the postmortem AD brain suggest significant involvement of inflammatory pathways in disease progression. PET imaging of AD patients with radio labeled tracers for microglial activation has confirmed these findings. Neuroinflammation involvement of microglia has also been observed in animal models used to study AD. However, the investigation of microglia in AD has been hampered by the lack of specific molecules for understanding microglial phenotype and function and equally as important there is the lack of therapeutic approaches that can target microglia and neuroinflammation. We hypothesize that neuroinflammation is an important target for developing new treatments for AD and that by dampening microglial neuroinflammation with our novel approach (nasal anti-CD3) we will be able to treat AD. We found that nasal anti-CD3 induces an anti-inflammatory immune response that attenuates neuroinflammation in brain-resident microglia cells. In new preliminary data, we found that nasal anti-CD3 improved short-term memory in an AD mouse model. We have found that nasally administered anti-CD3 localizes to cervical lymph nodes where it induces IL-10 secreting T cells that then migrate to the brain and suppress neuroinflammation in microglia. Thus, nasal administration of anti-CD3 is a unique and unexplored target for treating neuroinflammation in AD. The objective of this proposal is to obtain investigate the potential of nasal anti-CD3 to dampen microglia neuroinflammation as a treatment strategy in AD. Aim 1. Investigate the effect of nasal anti-CD3 on microglia phenotype and phagocytic function in aging. Nasal anti-CD3 will be given to 24-month old mice to assess the effect of treatment on in vivo microglia phagocytic function and restoration of microglia homeostasis. We will also measure the induction of IL-10- secreting T cells in the periphery of these mice, which will serve as an easily measured biomarker and on the role of IL-10 on mediating the effect of nasal anti-CD3 on microglia function in aging. Furthermore, we will use IL-10Rflox/floxCX3CR1Cre conditional knockout mice, which do not express the IL-10 receptor on microglia to investigate the role of IL-10 in microglia phagocytic properties as well as its ability to degrade Ab plaques. Aim 2. Investigate the effect of nasal anti-CD3 in the 3xTg AD model. We will perform detailed behavior analysis and neuropathology on 3xTg AD mice treated with nasal anti-CD3 to determine the biological effect and therapeutic potential of anti-CD3. Moreover, we will investigate the effect of nasal anti-CD3 treatment on microglial neuroinflammation in this mouse model of AD using high throughput RNA sequencing (Smart-Seq2).