Anti-inflammatory regulation of beta-amyloidosis

描述（由申请人提供）：本项目的主要目标是确定抗炎分子对阿尔茨海默病（AD）啮齿动物模型中神经炎症活动和β-淀粉样变性（A？聚集和沉积）的调节作用。我们以前的研究双转基因小鼠表达家族性AD突变体的淀粉样前体蛋白（APP）和胶质纤维酸性蛋白启动子驱动的小鼠CCL 2和APP转基因小鼠缺乏干扰素？I型受体证实趋化因子和促炎细胞因子在脑中β-淀粉样变性和小神经胶质增生的进展中起关键作用。因此，醋酸格拉替雷免疫，一种已知的抗炎疗法，减少了β-淀粉样变性，增强了神经发生，并改善了APP小鼠的认知功能。此外，特异性抗炎细胞因子（白细胞介素-4; IL-4、IL-10等）可以直接诱导小胶质细胞的抗炎和神经保护表型。因此，我们假设抗炎细胞因子（IL-4或IL-10等）可能诱导抑制体内神经炎症和β-淀粉样变性相关的认知功能障碍。我们的初步研究支持这一假设，因为慢性表达的中和CCL 2突变体（7 ND，缺乏前7个氨基酸序列）抑制小胶质细胞增生和A？寡聚体积累，并改善认知功能的双转基因小鼠（APP/PS1）表达APP和早老素-1（PS1）。利用腺相关病毒（AAV）介导的基因递送系统在APP/PS1小鼠中表达IL-4、IL-10、7 ND，在记忆功能障碍的症状前和症状后阶段，我们将提出以下问题：1）7 ND、IL-4或IL-10是否抑制星形/小胶质细胞增生？如果是，它是局限于海马区还是同时存在于皮层和海马区？2)IL-4或IL-10是否诱导树突样（CD 11 c+）小胶质细胞？如果是，它有神经保护作用吗？3)IL-4或IL- 10是否诱导炎症调节分子（CD 200、CD 200 R）？4)IL-4或IL-10能减轻β淀粉样变性吗？如果是，它是特异于致密斑块、弥散斑块还是A？寡聚体？5)IL-4或IL-10能促进神经发生吗？如果是这样，它是否伴随着增强的新合成的神经元或星形胶质细胞？6)IL-4或IL-10能促进突触发生吗？如果是，它是突触前还是突触后分子特有的？和7）7 ND、IL-4或IL-10是否在注射低剂量AAV后增强记忆形成？如果是，它在症状前和症状后阶段都有效吗？这一提议是重要的，因为据我们所知，7 ND、IL-4或IL-10基因递送的治疗功效从未在APP或APP/PS1小鼠体内测试过。这些方法可能对AD和其他神经退行性疾病的免疫治疗具有重要意义。公共卫生相关性：阿尔茨海默病（AD）是一种主要的神经系统疾病，在美国影响超过400万人，他们没有有效的治疗。本研究利用已建立的AD基因工程小鼠模型（APP/PS1小鼠），研究抗炎细胞因子（白细胞介素-4和10）和中和性趋化因子突变体（CCL 2）对APP/PS1小鼠β-淀粉样变性和认知功能的影响。这些方法可能对AD和其他神经退行性疾病具有治疗意义。

DESCRIPTION (provided by applicant): The over-arching goal of this project is to determine the regulatory role of anti-inflammatory molecules on neuroinflammatory activities and beta-amyloidosis (A¿ aggregation and deposition in brain) in rodent models of Alzheimer's disease (AD). Our previous studies on a double transgenic mouse expressing familial AD mutants of ¿-amyloid precursor protein (APP) and glial fibrillar acidic protein promoter-driven murine CCL2 and APP transgenic mice lacking interferon-? receptor type I demonstrated that chemokines and pro-inflammatory cytokines are critically involved in progression of beta-amyloidosis and microgliosis in brain. Accordingly, glatiramer acetate immunization, a known anti-inflammatory therapy, reduced beta-amyloidosis, enhanced neurogenesis, and improved cognitive function in APP mice. In addition, specific anti-inflammatory cytokines (interleukin-4; IL-4, IL-10, among others) can directly induce anti-inflammatory and neuroprotective phenotype of microglia. Thus, we hypothesize that anti-inflammatory cytokines (IL-4 or IL-10, among others) may induce suppression of neuroinflammation and beta-amyloidosis-related cognitive dysfunction in vivo. Our preliminary studies support this hypothesis, since chronic expression of neutralizing CCL2 mutant (7ND, lacking the first 7 amino acid sequence) suppresses microgliosis and A¿ oligomer accumulation, and improves cognitive function in a double transgenic mice (APP/PS1) expressing APP and presenilin-1 (PS1). Using adeno-associated virus (AAV)-mediated gene delivery system for expressing IL-4, IL-10, 7ND in APP/PS1 mice at pre- and post- symptomatic stages of memory dysfunction, we will ask the following questions: 1) Does 7ND, IL-4, or IL-10 suppress astro/microgliosis? If so, is it restricted to hippocampal region or both in cortex and hippocampus?; 2) Does IL-4 or IL-10 induce dendritic-like (CD11c+) microglia? If so, is it neuroprotective?; 3) Does IL-4 or IL- 10 induce inflammation regulatory molecules (CD200, CD200R)?; 4) Does IL-4 or IL-10 reduce beta- amyloidosis? If so, is it specific to compact plaques, diffuse plaques, or A¿ oligomers?; 5) Does IL-4 or IL-10 enhance neurogenesis? If so, is it accompanied with enhanced newly synthesized neurons or astrocytes? 6) Does IL-4 or IL-10 enhance synaptogenesis? If so, is it specific to presynaptic or postsynaptic molecules?; and 7) Does 7ND, IL-4 or IL-10 enhance memory formation after injection of lower doses of AAV? If so, is it effective in both pre-symptomatic and post-symptomatic stages? This proposal is significant, since to the best of our knowledge, therapeutic efficacy of 7ND, IL-4 or IL-10 gene delivery has never been tested in APP or APP/PS1 mice in vivo. These approaches may have significant implication for immunotherapy of AD and other neurodegenerative diseases. PUBLIC HEALTH RELEVANCE: Alzheimer's disease (AD) is a leading neurological disease that affects more than 4 million people in the US, who are left without effective therapy. Using the established the genetically engineered mouse model of AD (APP/PS1 mice), we will characterize the beneficial effect of anti-inflammatory cytokines (interleukin-4 and 10) and neutralizing chemokine mutant (for CCL2), on beta-amyloidosis and cognitive function inAPP/PS1 mice. These approaches may have therapeutic implication for AD and other neurodegenerative diseases.