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 是淀粉样蛋白(Aβ，Aβ)的胞外斑块沉积，它源于Aβ前体蛋白(APP)， 以及过度磷酸化的tau蛋白在细胞内的缠绕。此外，髓系细胞可能参与其中，例如 AD和TREM2蛋白之间的联系暗示。APP、tau和TREM2的表达受 非编码microRNA(MiRNA)，通过其mRNAs中的靶标。几种单核苷酸多态(SNP) 与AD关联。我们假设，通过改变miRNA，在UTR序列中自然产生SNPs 识别站点可以改变AD的风险和/或进展。我们建议在实验中测试miRNAs 证实可以改变AD相关的蛋白质水平，2)已经实验验证了这些mRNAs中的靶点，以及3)已经 人类阿尔茨海默病大脑中的扰动水平。我们提出了一项与阿尔茨海默病相关的系统性microRNA探索 以阐明这些变异的影响。我们的创新是评估 MiRNA中与疾病相关的功能活动靶点的自然发生的多态(SNP)。 新颖的仰慕平台通过连续的阶段理性地筛选候选多态。我们的 结果是从功能上验证已知的自然发生的多态在已确认的miRNA序列中 UTRs，采用三个具体目标(SA)。SA1：检验假说--自然发生的SNPs改变报告 对miRNA的反应。SA2：检验假说--自然产生的SNPs改变靶蛋白水平以应对 MiRNA治疗。SA3：检验假说-自然地，发生的SNPs改变目标蛋白水平以响应 MiRNA对人诱导多能干细胞(IPS)的治疗。 Ademire-Up平台要求SNP位于实验上已经被1)获得的miRNA靶序列内 确认改变了目标蛋白质水平；2)实验验证了序列；以及3)干扰了水平 与AD有关。然后，变异将与疾病基因组变异数据库交叉索引，以确定优先顺序 已报告的关联。符合这些标准的SNP将用于基于双报告程序的功能 化验。不同于野生型的SNPs活性将被用作模板来设计染色体突变 细胞培养。然后，通过这些阶段的SNPs将被用于来自临床来源的疾病特异性iPS细胞。 我们预计，这一过程的每一步都会产生一个“活跃”变种的收紧循环。那些变种 然后将在ADNI数据库中搜索仍显示与野生型应用程序显著差异的潜在应用程序 与AD相关表型的关联。 在R21的范围内，概念的证明将为严格和系统地探索 未统计的其他疾病相关基因非编码区序列中SNPs的功能效应。一个系统的 赞赏的是，测试多态对这种调节的影响的方法将加速更高的吞吐量 功能分析和更好地了解miRNA对疾病的影响或抗性的调节。