Salivary MicroRNAs as Biomarkers for Alcohol Dependence

描述(申请人提供)：酒精依赖(AD)是一种严重的公共卫生问题，其特征是酒精耐受性、身体依赖和无法控制酒精摄入量。它每年影响大约3.8%的美国成年人，每年造成巨大的经济损失。寻找有效的AD生物标志物是防治AD的关键。细胞培养和动物模型的研究表明，在转录后水平调节靶基因表达的小非编码microRNAs(MiRNAs)与慢性饮酒诱导的神经适应有关。新的证据支持miRNAs存在于细胞外空间或体液(如唾液、血清、尿液等)中，提示细胞外miRNAs是疾病诊断和预后的潜在生物标志物。到目前为止，关于人类AD患者miRNA表达变化的信息非常有限，目前还没有研究检测AD患者唾液中miRNA的表达变化。这项应用的目的是识别与AD相关的唾液miRNAs，并探索使用这些唾液miRNAs(结合与AD相关的SNPs)预测AD的可能性。中心假设是，长期饮酒会导致各种组织中miRNA表达的变化，这些变化的表达水平反映在miRNAs分泌到体液中，如唾液中。这一假说将通过追求三个具体目标来验证：(1)使用下一代测序(NGS)在非裔美国人和欧洲裔美国人中描述AD受试者唾液miRNA转录组的变化；(2)评估差异表达的miRNAs预测AD状态的能力；以及(3)结合AD相关唾液miRNA和AD相关SNPs(在我们最近的全基因组关联研究中确定)以改进AD的预测。我们希望发现一组与AD相关的唾液miRNAs和SNPs，它们可以作为AD的生物标志物。这种方法是创新的，因为AD患者唾液中miRNA转录组的变化将通过NGS技术进行检测，该技术在检测基因表达方面提供了前所未有的敏感性和特异性。此外，将miRNA表达和SNP基因分型数据结合起来有望大大提高AD的预测能力。我们的长期目标是使用已识别的AD相关唾液miRNAs作为新的诊断和预后生物标记物，并作为AD和AD相关疾病的潜在治疗靶点。这项拟议的研究具有重要意义，因为已识别的唾液miRNAs有望在监测AD的发展方面具有临床意义。此外，在这项拟议的研究中制定的研究策略可以应用于识别对其他物质使用障碍重要的细胞外miRNAs。

DESCRIPTION (provided by applicant): Alcohol dependence (AD) is a serious public health concern characterized by alcohol tolerance, physical dependence, and an inability to control one's alcohol intake. It affects approximately 3.8% of adult Americans each year, and causes substantial economic loss annually. Identification of effective biomarkers for AD is critical for A prevention and treatment. Studies of both cell culture and animal models have demonstrated that small non-coding microRNAs (miRNAs), which regulate the expression of their target genes at the post- transcriptional level, are implicated in chronic alcohol consumption-induced neuroadaptations. Emerging evidence supports the presence of miRNAs in extracellular spaces or body fluids (i.e., saliva, serum, urine, etc), suggesting that the extracellular miRNAs are potential biomarkers for disease diagnosis and prognosis. To date, there is very limited information on miRNA expression changes in human AD subjects, and no study is known to have examined miRNA expression alterations in the saliva of AD subjects. The objective of this application is to identify AD-associated salivary miRNAs and explore the possibility of using these salivary miRNAs (in combination with AD-associated SNPs) to predict AD. The central hypothesis is that chronic alcohol consumption induces altered miRNA expression in various tissues, and that these altered expression levels are reflected in the secretion of miRNAs into body fluids such as saliva. This hypothesis will be tested by pursuing three specific aims: (1) profile salivary miRNA transcriptome changes in AD subjects using next- generation sequencing (NGS) in both African Americans and European Americans; and (2) assess the ability of the differentially expressed miRNAs in predicting the status of AD; and (3) combine AD-associated salivary miRNA and AD-associated SNPs (identified in our recent genome-wide association studies) to improve the prediction of AD. We expect to discover a set of AD-associated salivary miRNAs and SNPs that can serve as AD biomarkers. The approach is innovative because miRNA transcriptome alterations in the saliva of AD subjects will be examined by NGS technology, which offers unprecedented sensitivity and specificity in the detection of gene expression. Moreover, integrating miRNA expression and SNP genotype data is expected to greatly improve the prediction of AD. Our long-term goal is to use the identified AD-associated salivary miRNAs as novel diagnostic and prognostic biomarkers and as potential therapeutic targets for AD and AD- related disorders. The proposed research is significant because the identified salivary miRNAs are expected to have clinical implications in monitoring the development of AD. Additionally, the research strategies developed in this proposed study can be applied to identify extracellular miRNAs that are important for other substance use disorders.