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

基本信息

批准号：
10653353
负责人：
Subodh Kumar
金额：
$ 24.9万
依托单位：
TEXAS TECH UNIVERSITY HEALTH SCIENCES CENTER AT EL PASO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-15 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10653353
关键词：
Address Age Alzheimer&apos s Disease Alzheimer&apos 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 insight miRNA expression profiling mind control mutant neuronal growth neurotransmission postsynaptic density protein synaptic function tau Proteins tau mutation tau-1 transcriptome sequencing

项目摘要

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 MicroRNA、阿尔茨海默病的突触损伤和认知能力下降项目概要/摘要我们研究的目的是确定突触体 microRNA (miRNA) 在阿尔茨海默病中的作用疾病（AD）的进展和发病机制是神经元最重要的区室。向邻近神经传递信号并维持大脑的健康突触功能。由突触小泡、突触蛋白、线粒体、神经递质受体、突触后组成最近，一些研究鉴定了 miRNA。突触、突触小泡和突触体的富集然而，突触相关的作用。 AD 进展中的 miRNA 尚未被探索。在我们研究的第一部分中，我们将确定这些 miRNA。 AD 中失调的突触体特异性 miRNA，进一步，我们将描述它们在 AD 中的作用进展和发病机制将从额叶皮质区域提取。从 AD (n=15) 和健康对照 (n=15) 的死后大脑中提取总 RNA。突触体部分并进行 miRNA 和 mRNA 测序处理。将使用 miRNA 模拟物和抑制剂方法以及突变体在体外对特定 miRNA 进行表征 APP 和 p-tau cDNA 构建体将研究突触体 miRNA 对 Aβ 和 p-tau 的影响。在第二部分中，我们将确定突触体 miRNA 在 Aβ 诱导的突触中的作用。将研究突触体 miRNA 对健康神经元和认知功能障碍的影响。具有 Aβ 诱导毒性的 AD 神经元我们将重点关注神经元处理、神经元生长和网络。使用 APP 原代神经元培养物改变突触体 miRNA 的发育和突触可塑性。将研究 APP 转基因 (TG) 和 APP 中疾病进展的表达和突触蛋白我们将执行具有各种认知功能的突触体特异性 miRNA 表达。 2、6、12 和 18 个月大 APP TG 和 APP KO 小鼠系相对于年龄匹配的 TG 阴性野生型 (WT) 小鼠，第三部分我们将研究突触体的作用。磷酸化 tau (p-tau) 中的 miRNA 对 AD 中突触和认知功能障碍的影响。突触体 miRNA 将研究 p-tau 诱导的神经元毒性、健康神经元加工、使用 tau 原代神经元培养的神经元生长和网络发育以及突触可塑性。将研究 Tau TG 和 Tau 疾病进展中突触体 miRNA 和突触蛋白的变化我们将执行具有各种认知行为和认知行为的突触体特异性 miRNA 表达。与 WT 小鼠相比，2、6、12 和 18 个月大的 tau TG 和 KO 小鼠的生化测试。将提供以下问题的答案 - 1) AD 突触处存在突触体 miRNA，2) 突触体- 与 AD 中的 Aβ 和 p-tau 病理相关的相关 miRNA，3) 整体突触功能、突触活性、评估突触体相关 miRNA 在 AD 中的治疗价值。