Synaptosomal MicroRNAs, Synaptic Damage and Cognitive Decline in Alzheimer's Disease

标题。突触体微RNA与阿尔茨海默病的突触损伤和认知功能减退 项目摘要/摘要 本研究的目的是确定突触体微RNAs(MiRNAs)在阿尔茨海默病中的作用 疾病(AD)的进展和发病机制。突触是神经元最重要的隔膜， 将信号传递给邻近的神经元，维持大脑健康的突触功能。Synapse领域是 由突触小泡、突触蛋白、线粒体、神经递质受体、突触后 密度蛋白和定位的小RNA和miRNAs。最近，几项研究确认了miRNAs 突触、突触小泡和突触小体的丰富。然而，突触相关的作用 在AD进展中的miRNAs是完全未知的。在我们研究的第一部分，我们将确定 突触体特异的miRNAs在AD中被解除调控，我们将进一步描述它们在AD中的作用 进展和发病机制。突触体组分将从大鼠额叶皮质提取 AD组和健康对照组各15例。总RNA将从 并对突触体组分进行miRNA和mRNA测序。失控的突触体- 将使用miRNA模拟和抑制(S)方法和突变体在体外对特定的miRNA进行表征 APP和p-tau基因的构建。将研究突触体miRNA对Aβ和p-tau的影响 诱导性毒性。在第二部分中，我们将确定突触体miRNA在β诱导的突触中的作用 以及AD患者的认知功能障碍。将研究突触体miRNAs对健康神经元和 A-β对AD神经元的毒性作用我们将专注于神经元处理、神经元生长和网络。 应用APP原代神经元培养的发育和突触可塑性。突触体miRNAs的改变 APP转基因(TG)和APP中的表达和突触蛋白将随着疾病的进展而研究 基因敲除(KO)小鼠。我们将以不同的认知方式执行突触体特异的miRNAs表达 2、6、12和18月龄APP TG和APP KO小鼠品系的行为和生化测试 年龄匹配的甘油三酯阴性野生型(WT)小鼠。在第三部分中，我们将研究突触体的作用 磷酸化tau(p-tau)中的miRNAs可诱导AD患者的突触和认知功能障碍。网络的影响 突触体miRNAs将研究p-tau对神经元的毒性，健康神经元的加工， 使用tau原代神经元培养的神经元生长和网络发育及突触可塑性。蚀变 将研究突触体miRNAs和突触蛋白与Tau、TG和Tau疾病进展的关系 Ko老鼠。我们将执行突触体特定的miRNAs表达，具有不同的认知行为和 2、6、12和18个月龄的tau TG和KO小鼠与WT小鼠的生化测试。我们的研究结果 将提供答案-1)突触体miRNAs存在于AD突触，2)突触体- 与AD的Aβ和p-tau病理相关的miRNA，3)整体突触功能，突触活性， 突触体相关miRNAs在阿尔茨海默病中的治疗价值

Title. Synaptosomal MicroRNAs, Synaptic Damage and Cognitive Decline in Alzheimer's Disease Project summary/abstract The purpose of our study is to determine the role of synaptosomal microRNAs (miRNAs) in Alzheimer's disease (AD) progression and pathogenesis. Synapses are the most important compartments of neuron that deliver signals to adjacent neuron and maintain healthy synaptic functions of the brain. Synapse territories are composed of synaptic vesicles, synaptic proteins, mitochondria, neurotransmitters receptors, postsynaptic density protein and localized small RNAs and miRNAs. Recently, several studies identified the miRNAs enrichments at synapse, synaptic vesicles and synaptosomes. However, the role of synapse-associated miRNAs in AD progression is completely unexplored. In the first part of our study, we will identify the synaptosome-specific miRNAs that are deregulated in AD, and further, we will characterize their roles in AD progression and pathogenesis. Synaptosomal fraction will be extracted from frontal cortex region of postmortem brains from AD (n=15) and healthy controls (n=15). Total RNA will be extracted from synaptosomal fraction and processed for miRNA and mRNA sequencing. The deregulated synaptosome- specific miRNAs will be characterized in vitro using miRNA mimics and inhibitor(s) approaches and mutant APP and p-tau cDNA constructs. Impact of synaptosomal miRNAs will be studied against Aβ and p-tau induced toxicities. In second part we will determine the role of synaptosomal miRNAs in Aβ induced synaptic and cognitive dysfunction in AD. The impact of synaptosomal miRNAs will be studied on healthy neurons and AD neurons with Aβ induced toxicities. We will focus on neuronal processing, neuronal growth and network development and synaptic plasticity using APP primary neuronal cultures. Alteration of synaptosomal miRNAs expression and synaptic proteins will be studied with disease progression in APP transgenic (TG) and APP knockout (KO) mice. We will execute the synaptosome-specific miRNAs expression with various cognitive behavioral and biochemical tests in 2-, 6-, 12- and 18-month-old APP TG and APP KO mice lines relative to age-matched TG negative wild-type (WT) mice. And in third part we will study the role of synaptosomal miRNAs in phosphorylated-tau (p-tau) induced synaptic and cognitive dysfunction in AD. The impact of synaptosomal miRNAs will be studied on p-tau induced toxicity in neurons, healthy neurons processing, neuronal growth and network development and synaptic plasticity using tau primary neuronal culture. Alteration of synaptosomal miRNAs and synaptic proteins will be studied with disease progression in Tau TG and Tau KO mice. We will execute the synaptosome-specific miRNAs expression with various cognitive behavioral and biochemical tests in 2-, 6-, 12- and 18-month-old tau TG and KO mice relative to WT mice. Our study outcome will provide the answers for - 1) presence of synaptosomal miRNAs at AD synapse, 2) synaptosome- associated miRNAs linked with Aβ and p-tau pathologies in AD, 3) Overall synapse function, synaptic activity, assessment of therapeutic value of synaptosome-associated miRNAs in AD.