Alzheimer's disease-linked microRNA Exploration of UTR Polymorphisms (AdmiRE-UP)

阿尔茨海默病相关 microRNA UTR 多态性探索 (AdMRE-UP)

基本信息

批准号：
10391153
负责人：
DEBOMOY K LAHIRI
金额：
$ 43.53万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
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：
2022-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10391153
关键词：
3&apos Untranslated Regions Age Alzheimer&apos s Disease Alzheimer&apos s disease brain Amyloid beta-Protein Amyloid beta-Protein Precursor Biological Assay Cell Culture Techniques Cell Line Cells Clinical Clone Cells Code Databases Deposition Disease Elements Engineering Future Gene Expression Gene Silencing Genes Genetic Databases Genetic Polymorphism Genetic Translation Genetic Variation Genomics Human In Situ Knowledge Link MAPT gene Measures Medical Genetics Messenger RNA Methods MicroRNAs Minor Mutate Mutation Myeloid Cells Neurofibrillary Tangles Neuronal Differentiation Neurons Outcome Outcome Measure Phenotype Post-Transcriptional Regulation Process Protein Precursors Proteins Protocols documentation Regulation Reporter Reporting Research Resistance Risk Senile Plaques Severities Single Nucleotide Polymorphism Site Source Synapses TREM2 gene Testing Tissues Translations Untranslated RNA Untranslated Regions Variant Work base brain tissue disease phenotype effective therapy endophenotype extracellular genomic data genomic variation hyperphosphorylated tau indexing induced pluripotent stem cell innovation mutant neuroinflammation novel rapid technique response screening sex tau Proteins tau-protein kinase trait

项目摘要

Effective treatments are urgently needed to cure or delay Alzheimer’s disease (AD). Major hallmarks of AD are extracellular plaque deposits of amyloid-β (Aβ) peptide, which is derived from the Aβ precursor protein (APP), and intracellular tangles of hyperphosphorylated tau protein. Further, myeloid cells may be involved, such as implied by connections between AD and the TREM2 protein. APP, tau, and TREM2 expression are regulated by non-coding microRNA (miRNA), via targets in their mRNAs. Several single-nucleotide polymorphisms (SNP) associate with AD. We hypothesize that naturally occurring SNPs within the UTR sequences by altering miRNA recognition sites can alter risk and/or progression of AD. We propose to test miRNAs that 1) are experimentally confirmed to alter AD-related protein levels, 2) have experimentally verified targets in these mRNAs, and 3) have perturbed levels in human AD brain. We propose a systematic Alzheimer’s disease-linked microRNA exploration of UTR polymorphisms (AdmiRe-Up) to elucidate effects of these variations. Our innovation is to assess naturally-occurring polymorphisms (SNPs) in miRNA target sites for functional activity in relation to disease. Novel AdmiRe-Up platform rationally winnows candidate polymorphisms through successive stages. Our outcome is to functionally validate known naturally-occurring polymorphisms in confirmed miRNA sequences in the UTRs, employing three specific aims (SA). SA1: Test hypothesis—naturally occurring SNPs alter reporter response to miRNA. SA2: Test hypothesis—naturally occurring SNPs alter target protein levels in response to miRNA treatment. SA3: Test hypothesis—naturally, occurring SNPs alter target protein levels in response to miRNA treatment in human induced pluripotent stem (iPS) cells. The AdmiRe-Up platform requires a SNP be within an miRNA target sequence that 1) has been experimentally confirmed to alter target protein levels; 2) was experimentally validated for sequence; and 3) has perturbed levels in association with AD. Variants will then be cross-indexed with disease genomic variation databases to prioritize already-reported associations. SNPs that pass these criteria will be used for dual-reporter based functional assays. SNPs activity that differ from wildtype will be used as templates to engineer chromosomal mutants in cell cultures. SNPs that pass these stages would then be used in disease-specific iPS cells from clinical sources. We expect each successive step of the process to produce a tightening circle of “active” variants. Those variants that still show significant difference from wildtype APP will then be searched for in the ADNI database for potential associations with AD-associated phenotypes. Within the limits of an R21, proof of the concept will lay a path for rigorous and systematic exploration of functional effects of SNPs in the UTR sequences of uncounted other disease-associated genes. A systematic method, AdmiRe-Up, of testing polymorphism effects on such regulation will accelerate higher-throughput functional analysis and greater understanding of miRNA modulation of disease effects or resistance.

迫切需要治愈或延迟阿尔茨海默氏病（AD）有效治疗。广告的主要标志是淀粉样蛋白β（Aβ）肽的细胞外斑块沉积物，它源自Aβ前体蛋白（APP），和高磷酸化tau蛋白的细胞内缠结。此外，可能涉及髓样细胞，例如 AD和TREM2蛋白之间的连接暗示。应用，tau和trem2的表达受非编码microRNA（miRNA），通过其mRNA中的靶标。几种单核苷酸多态性（SNP）与AD合作。我们假设通过改变miRNA在UTR序列中自然存在的SNP 识别地点可以改变AD的风险和/或进展。我们建议测试miRNA 1）在实验上是确认改变与AD相关的蛋白水平，2）在这些mRNA中具有实验验证的靶标，3）人类广告大脑中的干扰水平。我们提出了一个系统的阿尔茨海默氏病疾病与MicroRNA探索 UTR多态性（欣赏）以阐明这些变化的影响。我们的创新是评估 miRNA靶位点中的自然出现多态性（SNP），用于与疾病有关的功能活性。新颖的欣赏平台通过成功的阶段合理地赢得了候选多态性。我们的结果是在功能上验证在确认的miRNA序列中验证已知的已知天然多态性 UTR，采用三个特定目标（SA）。 SA1：测试假设 - 自然发生的SNP改变报告基因对miRNA的反应。 SA2：检验假设 - 自然发生的SNP改变了目标蛋白水平 miRNA处理。 SA3：检验假设 - 自然而然地发生SNP会改变目标蛋白水平人类诱导多能干（IPS）细胞中的miRNA处理。欣赏的平台要求SNP在miRNA目标序列内，即1）已在实验中已确认改变靶蛋白水平； 2）对序列进行了实验验证； 3）具有干扰水平与AD联合。然后，变体将与疾病基因组变异数据库进行交叉索引，以优先考虑已经报告的协会。传递这些标准的SNP将用于基于双重复器的功能测定。与WildType不同的SNP活性将用作工程染色体突变体的模板细胞培养。然后，通过这些阶段的SNP将用于临床来源的疾病特异性IPS细胞中。我们期望该过程的每个成功步骤都会产生一个“主动”变体的收紧圆圈。那些变体然后将在ADNI数据库中搜索与Wildtype应用程序显着差异的潜力与广告相关表型的关联。在R21的范围内，该概念的证明将为严格而系统的探索提供一条途径 SNP在未成立的其他疾病相关基因的UTR序列中的功能效应。系统测试多态性对这种法规的测试效应的方法，令人钦佩功能分析以及对miRNA调节疾病效应或抗药性的更多了解。