Biomarker Expression and Regulatory Haplotypes in Alzheimer's Disease

项目概要/摘要 阿尔茨海默病（AD）死后脑组织的特征性表现是神经元变性 以及大量的淀粉样蛋白沉积物（A <$42）（淀粉样蛋白前体的裂解产物 由APP基因编码的蛋白）和tau（由MAPT基因编码）。脑脊液（CSF）A <$42和 tau蛋白水平可以预测AD，但在区分AD与其他神经退行性疾病方面的可靠性有限。 疾病载脂蛋白（APOE）<$4等位基因和年龄是目前唯一与糖尿病密切相关的因素。 晚发性AD我们对AD遗传方面的理解存在很大差距，可能包括其他遗传因素。 影响迟发性AD的发病年龄和表型表达的因素。个体遗传学发现（非 与复杂疾病（如AD）相关的同义SNP不太可能完全解释AD的实质性差异。 遗传变异对疾病发病机制影响。多层次的病因可能是复杂的 疾病，并且可以包括影响转录调节的基因内和周围的多个基因座 和转录后，强调需要综合评估大的遗传区域， 与蛋白质生物标志物水平的相关性作为预测疾病风险的手段。该提案的重点是 大基因区域周围和内部的多个遗传基因座起调节基因表达的作用 以AD特异性方式表达。在本研究的指导阶段（K99），第一个目标是 寻找APOE、APP和MAPT周围和内部的多个基因座或SNP（单倍型）组合 与CSF和PMB中的表达水平相关的基因。候选遗传和蛋白质生物标志物将 扩展到APOE、APP和MAPT基因之外，以包括可能与以下生物学相关的其他基因： 神经退行性疾病第二个目的是证明假定的调节单倍型在功能上 通过利用基因组DNA影响表达，所述基因组DNA含有特定的推定调节单倍型，作为活性成分， 报告基因和小基因测定中基因调控位点。在独立阶段（R 00），最终目标是 测试调节单倍型在区分不同AD表型和AD表型之间的可靠性， 和其他神经退行性疾病。总的来说，这些提议的实验是独一无二的，因为它们 除了核心启动子基因座和生物标志物表达水平之间的简单相关性之外， 遗传、统计和功能技术的组合，以评估多个基因座在 推测AD相关基因表达的远端调控元件，以发现预测AD的单倍型。的 该K99/R 00应用程序的研究和职业发展部分将提供必要的培训 申请人成为一个成功的独立调查员谁可以整合这些技术， 提高我们对神经退行性疾病风险的认识。

Project Summary/Abstract The characteristic findings in Alzheimer's disease (AD) post-mortem brain (PMB) are degeneration of neurons together with extensive amounts of amyloid deposits (A¿42) (a cleavage product of the amyloid precursor protein encoded by the APP gene) and tau (encoded by the MAPT gene). Cerebrospinal fluid (CSF) A¿42 and tau levels can predict AD but have a limited reliability in discriminating AD from other neurodegenerative diseases. The apolipoprotein (APOE) ¿4 allele and age are currently the only factors strongly associated with late onset AD. The large gaps in our understanding of the genetic aspects of AD may include additional genetic factors that impact age of onset and phenotypic expression of late onset AD. Individual genetic findings (non- synonomous SNPs) associated with complex diseases, such as AD, are unlikely to fully explain the substantial impact of genetic variation on disease pathogenesis. Multilevel etiologic factors are likely to underlie complex diseases and may include multiple loci within and surrounding a gene that influence regulation of transcription and post-transcription, emphasizing the need for integrative evaluation of large genetic regions and correlations with protein biomarker levels as a means for predicting disease risk. This proposal focuses on the overall hypothesis that multiple genetic loci surrounding and within large gene regions act to regulate gene expression in an AD specific manner. During the mentored phase (K99) of this investigation the first aim is to find multiple loci or combinations of SNPs (haplotypes) surrounding and within the APOE, APP and MAPT genes that correlate with expression levels in CSF and PMB. Candidate genetic and protein biomarkers will expand beyond APOE, APP and MAPT genes to include other genes likely to be biologically relevant to neurodegenerative disease. The second aim is to demonstrate that putative regulatory haplotypes functionally impact expression by utilizing genomic DNA, containing a particular putative regulatory haplotype, as the active site of gene regulation in reporter and minigene assays. During the independent phase (R00), the final aim is to test regulatory haplotypes for their reliability in discerning between different AD phenotypes and between AD and other neurodegenerative diseases. Collectively, these proposed experiments are unique because they go beyond the simple correlation between core promoter loci and biomarker expression levels by using a combination of genetic, statistical and functional techniques to evaluate the influence of multiple loci within putative distant regulatory elements on AD relevant gene expression to find haplotypes that predict AD. The research and career development components of this K99/R00 application will provide the necessary training for the applicant to become a successful independent investigator who can integrate these techniques to improve our understanding of neurodegenerative disease risk.