microRNA (miRNA) signaling in Alzheimer's disease(AD)

阿尔茨海默病(AD)中的microRNA(MiRNA)信号传导 通过广泛的基于miRNA和DNA的表达阵列、LED-Northern、ELISA、 基于西方、免疫学和生物信息学的分析，我们发现了高度 核因子-kB敏感、上调的促炎microRNAs(MiRNAs)的相互作用网络 以及它们下调的信使RNA(MRNAs)靶标和这些mRNAs的蛋白质 在散发性阿尔茨海默病(AD)脑中编码。这些致病因子的上调 对应激人类miRNAs及其下调的mRNAs靶标进行了进一步分析 原代培养的脑细胞和高表达5xFAD淀粉样蛋白的转基因小鼠。 通过miRNA和mRNA丰度分析，miRNA-mRNA互补作图， 联合能量(EA)指数、酶联免疫吸附试验和Western分析可以解释大部分 通过分析AD过程中的显著干扰来观察AD过程的神经病理学特征 选择性miRNA-mRNA信号转导。我们的假设是存在一个至少6口人的小家庭 AD脑内关键的促炎miRNAs负责靶向和下调a 一组致病信使核糖核糖核酸(信使核糖核酸)负责淀粉样变的靶标 伴随着突触发生缺陷的病理和神经炎症，先天的- 免疫、吞噬和Aβ42肽清除进行性损害。 我们之前的5年NIA R01的续订将调查 6个促炎症、致病的miRNAs在散发性AD脑中上调，在应激状态下 5xFAD小鼠脑内原代培养的人脑细胞。6个上调的PRO- 需要详细研究的炎症性microRNA有miRNA-7、miRNA-9、miRNA-34a、miRNA-125b、 MiRNA-146a和miRNA-155。应激源将是在老化的AD大脑中发现的那些 -包括活性氧(ROS)、促炎细胞因子IL-1β和肿瘤坏死因子α 和Aβ42肽。具体目标1将分析这些因素在中等至 晚期散发性阿尔茨海默病和唐氏综合症(DS)脑；特定目标2将分析 这些因素在应激人脑细胞中的作用，即在神经元-神经胶质原代细胞联合 培养；特定目标3将分析这些因素在5xFAD淀粉样蛋白过多中的贡献。 表达转基因小鼠品系。我们将特别强调最UP的研究- 受调控的miRNA：miRNA-7、miRNA-9、miRNA 34a和miRNA-146a及其靶向 UBE2A(泛素结合酶蛋白)和TREM2(触发受体，表达于 小胶质细胞)信号在阿尔茨海默病脑内，以及在体外和体内的AD模型。我们还将 分析选择性核因子-kB抑制剂和抗miRNA(AM)策略在 在这个系统中恢复动态平衡。我们的长期目标是治疗 这些miRNA调控的表观遗传途径为临床提供了一种有效的治疗方法 AD的早期管理。

microRNA (miRNA) signaling in Alzheimer's disease (AD) Through extensive miRNA- and DNA-based expression array-, LED-Northern-, ELISA, Western-, immunological and bioinformatics-based analysis we have discovered a highly interactive network of NF-kB sensitive, up-regulated pro-inflammatory microRNAs (miRNAs) and their down-regulated messenger RNA (mRNA) targets and the proteins these mRNAs encode in sporadic Alzheimer's disease (AD) brain. The up-regulation of these pathogenic miRNAs and their down-regulated mRNA targets has been further analyzed in stressed human brain cells in primary culture and in 5xFAD amyloid over-expressing transgenic mouse lines. Through miRNA and mRNA abundance analysis, miRNA-mRNA complementarity mapping, association energy (EA) indexing, ELISA and Western analysis we can explain much of the observed neuropathology characteristic of the AD process by analyzing significant disruptions in selective miRNA-mRNA signaling. Our hypothesis is that there exists a small family of at least 6 critical pro-inflammatory miRNAs in AD brains responsible for targeting and down-regulating a group of pathogenic messenger RNA (mRNA) targets responsible for amyloidogenesis, tau pathology and neuroinflammation with accompanying deficits in synaptogenesis, innate- immunity, phagocytosis and a progressive impairment in Aβ42 peptide clearance. This renewal of our previous 5 year NIA R01 will investigate the integrated actions of this group of 6 pro-inflammatory, pathogenic miRNAs up-regulated in sporadic AD brain, in stressed human brain cells in primary culture in the brains of 5xFAD mice. The 6 up-regulated pro- inflammatory microRNAs to be studied in detail are miRNA-7, miRNA-9, miRNA-34a, miRNA-125b, miRNA-146a and miRNA-155. Stressors will be those encountered as are found in aging AD brain – these include reactive oxygen species (ROS), the pro-inflammatory cytokines IL-1β and TNFα and Aβ42 peptides. Specific Aim 1 will analyze the contribution of these factors in moderate-to- advanced sporadic AD and Down's syndrome (DS) brain; Specific Aim 2 will analyze the contribution of these factors in stressed human brain cells, i.e. in neuronal-glial primary cell co- cultures; Specific Aim 3 will analyze the contribution of these factors in 5xFAD amyloid over- expressing transgenic mouse lines. We will specifically accentuate the study of the most up- regulated miRNAs: miRNA-7, miRNA-9, miRNA 34a and miRNA-146a and their targeted disruption of UBE2A (ubiquitin conjugase protein) and TREM2 (triggering receptor expressed in microglial cells) signaling in AD brain, and in in vitro and in vivo AD models. We will also analyze the applicability of selective NF-kB inhibitors and anti-miRNA (AM) strategies in restoring homeostasis in this system. Our long term goal is the therapeutic manipulation of these miRNA-regulated epigenetic pathways to provide an efficacious treatment for the clinical management of AD at an early stage.