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

阿尔茨海默病中的突触体 MicroRNA、突触损伤和认知能力下降

• 批准号: 10260431
• 负责人: Subodh Kumar
• 金额: $ 9.51万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10260431
• 关键词: Address; Age; Alzheimer' s Disease; Alzheimer' s disease brain; Amyloid beta-Protein; Autopsy; Behavior; Behavioral; Biochemical; Brain; Cognitive; Complementary DNA; Complex; Data Analyses; Dendritic Spines; Development; Disease Progression; Functional disorder; Genomics; Goals; Hippocampus (Brain); Impaired cognition; In Vitro; Knockout Mice; Lead; Link; Mediating; Messenger RNA; MicroRNAs; Microarray Analysis; Mitochondria; Molecular; Mus; Neurodegenerative Disorders; Neurons; Neurotransmitter Receptor; Neurotransmitters; Outcome; Outcome Study; Pathogenesis; Pathology; Physiological; Process; Property; Protein Biosynthesis; Proteins; Proteomics; Publishing; Quantitative Reverse Transcriptase PCR; RNA; Regulation; Reporting; Research; Role; Signal Transduction; Small RNA; Synapses; Synaptic Vesicles; Synaptic plasticity; Synaptosomes; Testing; Therapeutic; Toxic effect; Transcript; Transgenic Mice; Transgenic Organisms; United States National Institutes of Health; Wild Type Mouse; base; cognitive function; design; differential expression; disorder control; frontal lobe; improved; inhibitor/antagonist; insight; miRNA expression profiling; mind control; mutant; neuronal growth; neurotransmission; postsynaptic density protein; synaptic function; tau Proteins; tau mutation; tau-1; transcriptome sequencing

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.

标题.突触体microRNAs、突触损伤与阿尔茨海默病认知功能减退 项目概要/摘要 我们研究的目的是确定突触体微小RNA（miRNA）在阿尔茨海默病中的作用 疾病（AD）进展和发病机制。突触是神经元中最重要的部分， 将信号传递到相邻的神经元并维持大脑的健康突触功能。突触的领地是 由突触囊泡、突触蛋白、线粒体、神经递质受体、突触后组成 密度蛋白和定位的小RNA和miRNA。最近，几项研究鉴定了miRNAs， 在突触、突触囊泡和突触体处富集。然而，突触相关的作用 miRNAs在AD进展中的作用完全未被探索。在我们研究的第一部分，我们将确定 突触体特异性miRNA在AD中的失调，进一步，我们将描述它们在AD中的作用 进展和发病机制。突触体部分将从额叶皮质区域提取 来自AD（n=15）和健康对照（n=15）的死后脑。总RNA将从 突触体部分并加工用于miRNA和mRNA测序。去调节的突触体- 特异性miRNA将使用miRNA模拟物和抑制剂方法在体外表征，并且突变体 APP和p-tau cDNA构建体。将研究突触体miRNA对Aβ和p-tau的影响 诱发毒性。在第二部分中，我们将确定突触体miRNAs在Aβ诱导的突触形成中的作用。 和认知功能障碍。将研究突触体miRNA对健康神经元的影响， Aβ可诱导AD神经元毒性。我们将集中在神经元加工，神经元生长和网络 使用APP原代神经元培养物的发育和突触可塑性。突触体miRNA的改变 将在APP转基因（TG）和APP中研究突触蛋白的表达和疾病进展。 敲除（KO）小鼠。我们将用不同的认知方法来执行突触体特异性miRNAs的表达， 在2、6、12和18月龄APP TG和APP KO小鼠系中进行的行为和生化试验， 年龄匹配的TG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蛋白（p-tau）中的miRNA诱导AD中的突触和认知功能障碍。的影响 将研究突触体miRNAs对神经元中p-tau诱导的毒性，健康神经元加工， 使用tau原代神经元培养物的神经元生长和网络发育以及突触可塑性。改变 突触体miRNA和突触蛋白的表达将随着Tau TG和Tau KO小鼠。我们将在不同的认知行为和认知功能的情况下进行突触体特异性miRNAs的表达， 在2月龄、6月龄、12月龄和18月龄的tau TG和KO小鼠中相对于WT小鼠的生物化学测试。我们的研究结果 将为以下问题提供答案：1）AD突触中突触体miRNA的存在，2）突触体- 与AD中的Aβ和p-tau病理学相关的相关miRNA，3）总体突触功能，突触活性， 评估突触体相关miRNA在AD中的治疗价值。