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)

• 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
• 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' Untranslated Regions; Age; Alzheimer' s Disease; Alzheimer' 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

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.

迫切需要有效的治疗方法来治愈或延缓阿尔茨海默病（AD）。AD的主要特征 是β淀粉样蛋白（Aβ）肽的细胞外斑块沉积，A β肽来源于Aβ前体蛋白（APP）， 和过度磷酸化的tau蛋白的细胞内缠结。此外，可能涉及骨髓细胞，例如 AD和TREM 2蛋白之间的联系暗示了这一点。APP、tau和TREM 2的表达受 非编码microRNA（miRNA），通过其mRNA中的靶点。几种单核苷酸多态性（SNP） 与AD合作我们假设，通过改变miRNA， 识别位点可以改变AD的风险和/或进展。我们建议测试以下miRNAs：1）实验上 证实改变AD相关蛋白质水平，2）实验验证了这些mRNA中的靶点，3） 人类AD大脑中的干扰水平。我们提出了一个系统的阿尔茨海默病相关的microRNA探索 的UTR多态性（AdmiRe-Up），以阐明这些变化的影响。我们的创新是评估 miRNA靶位点中天然存在的多态性（SNP）与疾病相关的功能活性。 新的AdmiRe-Up平台通过连续的阶段合理地筛选候选多态性。我们 结果是在功能上验证已证实的miRNA序列中已知的天然存在的多态性， UTR，采用三个具体目标（SA）。SA 1：测试假设-天然存在的SNP改变报告基因 对miRNA的反应SA 2：测试假设-天然存在的SNP改变靶蛋白水平， miRNA治疗。SA 3：测试假设-自然发生的SNP改变靶蛋白水平， 人诱导多能干细胞（iPS）中的miRNA治疗。 AdmiRe-Up平台要求SNP位于1）已经实验证实的miRNA靶序列内 证实改变靶蛋白水平; 2）实验验证序列; 3）具有扰动水平 与AD有关。然后将变异与疾病基因组变异数据库交叉索引， 已经报道的协会。通过这些标准的SNP将用于基于双报告子的功能性分析。 分析。与野生型不同的SNP活性将被用作模板，以工程化染色体突变体， 细胞培养然后，通过这些阶段的SNP将用于来自临床来源的疾病特异性iPS细胞。 我们预计，该过程的每一个连续步骤都会产生一个“活跃”变体的收紧圈。那些变体 然后在ADNI数据库中搜索仍然显示出与野生型APP显著差异的基因， 与AD相关表型的关联。 在R21的限制内，概念的证明将为严格和系统地探索 在无数其他疾病相关基因的UTR序列中SNP的功能效应。一个系统 一种测试多态性对这种调节的影响的方法AdmiRe-Up将加速更高的吞吐量 功能分析和更好地理解miRNA对疾病效应或抗性的调节。