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-β (Aβ) 肽的细胞外斑块沉积物，该肽源自 Aβ 前体蛋白 (APP)， 以及过度磷酸化 tau 蛋白的细胞内缠结。此外，骨髓细胞也可能参与其中，例如 AD 和 TREM2 蛋白之间的联系暗示了这一点。 APP、tau 和 TREM2 表达受调节 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 相关。我们假设 UTR 序列中自然存在的 SNP 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 miRNA：1) 通过实验 已证实可以改变 AD 相关蛋白水平，2) 已经通过实验验证了这些 mRNA 中的靶点，并且 3) 人类 AD 大脑中的扰动水平。我们提出了与阿尔茨海默病相关的 microRNA 的系统探索 UTR 多态性 (AdmiRe-Up) 来阐明这些变异的影响。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。 新颖的 AdmiRe-Up 平台通过连续阶段合理筛选候选多态性。我们的 结果是在功能上验证已确认的 miRNA 序列中已知的自然发生的多态性 UTR，采用三个具体目标（SA）。 SA1：检验假设——自然发生的 SNP 改变报告基因 对 miRNA 的反应。 SA2：测试假设——天然存在的 SNP 会改变靶蛋白水平以响应 miRNA 治疗。 SA3：检验假设——自然地，发生的 SNP 会改变靶蛋白水平以响应 人诱导多能干 (iPS) 细胞中的 miRNA 处理。 AdmiRe-Up 平台要求 SNP 位于 miRNA 目标序列内，1) 已通过实验 证实可以改变目标蛋白水平； 2) 序列经过实验验证； 3) 水平受到干扰 与 AD 相关。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 已经报告的协会。通过这些标准的 SNP 将用于基于双报告基因的功能 化验。与野生型不同的 SNP 活性将被用作模板来设计染色体突变体 细胞培养物。通过这些阶段的 SNP 将被用于临床来源的疾病特异性 iPS 细胞。 我们期望该过程的每个连续步骤都会产生“活跃”变体的紧缩循环。那些变体 然后将在 ADNI 数据库中搜索与野生型 APP 仍显示出显着差异的潜在 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的探索铺平道路 未计数的其他疾病相关基因的 UTR 序列中 SNP 的功能影响。一个系统的 测试多态性对此类调节的影响的方法 AdmiRe-Up 将加速更高的通量 功能分析并更好地了解 miRNA 对疾病影响或抵抗力的调节。