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: 9977483
• 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/9977483
• 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。阿尔茨海默氏病的突触体microRNA，突触损伤和认知能力下降 项目摘要/摘要 我们研究的目的是确定突触体microRNA（miRNA）在阿尔茨海默氏症中的作用 疾病（AD）进展和发病机理。突触是神经元中最重要的隔室 向相邻神经元传递信号并保持大脑的健康突触功能。突触领域是 由突触蔬菜，突触蛋白，线粒体，神经递质受体，突触后组成 密度蛋白以及局部的小RNA和miRNA。最近，一些研究确定了miRNA 突触，突触蔬菜和突触体的富集。但是，突触相关的作用 广告进展中的miRNA是完全出乎意料的。在我们研究的第一部分中，我们将确定 在AD中放松管制的突触体特异性miRNA，此外，我们将表征它们在AD中的作用 进展和发病机理。突触体分数将从额叶皮质区域提取 AD（n = 15）和健康对照（n = 15）的尸体大脑。总RNA将从 突触体馏分，用于miRNA和mRNA测序。放松管制的突触体 - 特定的miRNA将在体外使用miRNA模拟物和抑制剂方法和突变体进行体外表征 App和P-TAU cDNA构建体。突触体miRNA的影响将对Aβ和P-TAU进行研究 诱导的毒性。在第二部分中，我们将确定突触小体miRNA在Aβ诱导的突触中的作用 和AD中的认知功能障碍。突触体miRNA的影响将研究健康的神经元和 Aβ诱导毒性的AD神经元。我们将专注于神经元处理，神经元增长和网络 使用应用原发性神经元培养物的开发和突触可塑性。突触小体miRNA的改变 表达和突触蛋白将研究APP转基因（TG）和APP中的疾病进展 敲除（KO）小鼠。我们将用各种认知执行突触体特异性miRNA表达 相对于2个，6-，6，12个月和18个月大的APP TG和APP KO小鼠线的行为和生化测试相对于 年龄匹配的TG负野生型（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）诱导AD的突触和认知功能障碍的miRNA。的影响 突触体miRNA将在p-TAU诱导的神经元，健康神经元加工的毒性上进行研究， 神经元的生长和网络发展和突触可塑性使用TAU原发性神经元培养。改造 突触体miRNA和突触蛋白的疾病将在tau tg和tau中进行疾病进展。 KO老鼠。我们将使用各种认知行为和 相对于WT小鼠，在2个，6，12个月和18个月大的Tau Tg和Ko小鼠中进行生化测试。我们的研究结果 将提供-1）AD突触时存在突触体miRNA的答案，2）突触体 - 与Aβ和P-TAU病理相关的相关miRNA，3）总体突触功能，突触活动， 在AD中与突触体相关的miRNA的治疗价值评估。