Neurobiological Role of MicroRNA in Alzheimer's

MicroRNA 在阿尔茨海默病中的神经生物学作用

• 批准号: 10901008
• 负责人: DEBOMOY K LAHIRI
• 金额: $ 62.73万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10901008
• 关键词: 3' Untranslated Regions; Age; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease diagnosis; Alzheimer’s disease biomarker; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Anatomy; Apoptosis; Apoptotic; Autopsy; Biological; Biological Assay; Biological Markers; Brain; Brain region; Cell Culture Techniques; Cell Death; Cell Survival; Cell physiology; Cells; Clinical Trials; Correlation Studies; Databases; Dementia; Diagnostic; Disease; Disease Progression; Failure; Genes; Genotype; Goals; Growth; Health; Human; IL6 gene; Impairment; Interleukin-1; Lewy Body Disease; Link; MAPT gene; Measures; Mediating; Messenger RNA; MicroRNAs; Modeling; Morphology; Nerve Degeneration; Neurobiology; Neurodegenerative Disorders; Neuroglia; Neurons; Organism; Outcome; Pathogenesis; Pathology; Pattern; Physiological; Play; Progress Reports; Proteins; RE1-silencing transcription factor; Regulation; Regulatory Pathway; Reporter; Reporting; Research; Rest; Role; Sampling; Site; Symptoms; Testing; Transfection; Translating; Translations; Untranslated Regions; Work; age related; brain cell; cell type; cohort; cost; cytokine; endophenotype; gene product; genomic data; improved; induced pluripotent stem cell; innovation; mRNA Translation; member; mild cognitive impairment; neuroinflammation; neuronal survival; neurotoxicity; new therapeutic target; novel; novel diagnostics; novel therapeutic intervention; prevent; risk prediction; sex; synuclein; tau Proteins; tau-1; tool; translational study; treatment response

Our goal is to identify mechanisms leading to neurodegeneration in Alzheimer's disease (AD). We will study the regulation of amyloid-β precursor protein (APP), microtubule-associated protein tau (MAPT), −Synuclein (SNCA), and RE1-Silencing Transcription Factor (REST). This proposal will test miR153 as a 'regulator of the regulators'. We have established that miR153 regulates the translation of important AD-associated mRNAs for APP, SNCA, and REST via the untranslated region (UTR). We propose testing the hypothesis that miR153 serves as a nexus regulator of multiple AD-related proteins and control neuronal survival. This proposal will significantly advance the understanding of miR153's role in regulating proteins related to neurodegeneration. SA1. Analyze miR153 interacts with target sites in REST-153 network (R-153net) partner mRNAs. The proteins REST, APP, tau, IL1α, and IL6 play critical roles in AD. A "master regulator" that controls such protein levels could be a vital tool to prevent AD-related neurotoxicity. We will test the activity of miR153 on multiple mRNA-fused reporter clones based on the mRNA sequence of selected genes. We expect miR153 treatments will silence predicted target sites, and miRNA treatment will further alter protein levels in cell cultures. SA2. Test miR153 perturbs expression of the R-153net and cell health and growth. We will define the neurobiology of miR153's interactions, including reactive oxidizing species (ROS). We will establish effects on native protein levels and cell survival. Identifying the roles of miRNA-mediated regulation of the R-153net will elucidate miRNA-dependent mechanisms to enhance cell vitality. We expect that miR153 interacts with overall cell health, and these studies will reveal mechanisms for miR153 function in cell survival. SA3. Measure miR153 treatment alters the R-153net in induced pluripotent stem cells (iPSC). We will test the effects of miR153 treatment in differentiated iPSC from normal and AD donors. We expect that miR153 will alter R-153net levels and cell vitality and morphology. "Forking" iPSC cultures into neurons, glia, and mixed-type cell induction will allow for explicitly measuring effects in each of the major brain cell types. SA4. Assess miR153 is dysregulated in AD stage and brain region-specific manners. We will test and model miR153, mRNA, and protein levels in AD, MCI, and control human brain samples, along with APOE, sex, and age vs. disease progression. We expect levels of miR153 and R-153net proteins and mRNAs to vary in progression-dependent manners. Our results will predict the risk associated with R-153net member levels. SA5. Identify SNPs near MIR153 genes associate with altered CSF Aβ and phospho-tau levels. We will identify the effects of SNPs close to the MIR153-1 and -2 genes. We will investigate large-scale genomic data for SNP associations with AD endophenotypes, which will provide predictive AD biomarkers. Our studies in cell cultures, iPSCs, miRNA expression in human brains, and identifying SNPs near MIR153 genes will establish their relationships to AD and lead to novel diagnostic and therapeutic strategies.

我们的目标是确定导致阿尔茨海默病（AD）神经退行性变的机制。我们将研究 淀粉样蛋白-β前体蛋白（APP）、微管相关蛋白tau（MAPT）、β-突触核蛋白 （SNCA）和RE 1-沉默转录因子（REST）。这项提案将测试miR 153作为“调节剂”的作用。 监管机构的。我们已经确定miR 153调节重要AD相关mRNA的翻译， APP、SNCA和REST通过非翻译区（UTR）。我们提出检验miR 153 作为多种AD相关蛋白的连接调节剂并控制神经元存活。这项建议会 这一发现极大地推进了对miR 153在调节与神经变性相关的蛋白质中的作用的理解。 SA 1.分析miR 153与REST-153网络（R-153 net）伴侣mRNA中的靶位点相互作用。的 蛋白REST、APP、tau、IL 1 α和IL 6在AD中起关键作用。控制这种蛋白质的“主调节器” 水平可能是预防AD相关神经毒性的重要工具。我们将在多个细胞上测试miR 153的活性。 基于所选基因的mRNA序列的mRNA融合报告基因克隆。我们希望miR 153治疗 将沉默预测的靶位点，并且miRNA处理将进一步改变细胞培养物中的蛋白质水平。 SA 2.测试miR 153干扰R-153 net的表达以及细胞健康和生长。我们将定义 miR 153相互作用的神经生物学，包括活性氧化物质（ROS）。我们将建立对 天然蛋白质水平和细胞存活。确定miRNA介导的R-153 net调节的作用将 阐明miRNA依赖性机制以增强细胞活力。我们预计miR 153与总体 这些研究将揭示miR 153在细胞存活中的作用机制。 SA 3.测量miR 153处理改变诱导多能干细胞（iPSC）中的R-153 net。我们将 测试miR 153处理在来自正常和AD供体的分化iPSC中的作用。我们希望miR 153 将改变R-153 net水平以及细胞活力和形态。将iPSC培养物“分叉”成神经元、神经胶质和 混合型细胞诱导将允许明确地测量在每种主要脑细胞类型中的作用。 SA 4.评估miR 153在AD阶段和脑区域特异性方式中的失调。我们将测试和 AD、MCI和对照人脑样品中的模型miR 153、mRNA和蛋白水平，沿着APOE， 性别和年龄与疾病进展。我们预计miR 153和R-153 net蛋白和mRNA的水平会有所不同， 以依赖于进展的方式。我们的研究结果将预测与R-153 net成员水平相关的风险。 SA 5.鉴定与CSF Aβ和磷酸化tau水平改变相关的MIR 153基因附近的SNP。我们 将确定与MIR 153 -1和MIR 153 - 2基因接近的SNP的影响。我们将研究大规模的基因组 SNP与AD内表型相关的数据，这将提供预测性AD生物标志物。我们的研究 在细胞培养中，iPSC，人脑中的miRNA表达，以及识别MIR 153基因附近的SNP 将建立他们与AD的关系，并导致新的诊断和治疗策略。