Salivary MicroRNAs as Biomarkers for Alcohol Dependence

描述（由申请人提供）：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、身体依赖性和无法控制酒精摄入。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。识别有效的AD生物标志物对于AD的预防和治疗至关重要。细胞培养和动物模型的研究表明，在转录后水平调节其靶基因表达的小的非编码microRNA（miRNA）与慢性酒精消耗诱导的神经适应有关。新出现的证据支持细胞外空间或体液中存在miRNA（即，唾液、血清、尿液等），表明细胞外miRNA是用于疾病诊断和预后的潜在生物标志物。迄今为止，关于人类AD受试者中miRNA表达变化的信息非常有限，并且没有已知的研究已经检查了AD受试者唾液中miRNA表达的改变。本申请的目的是鉴定AD相关的唾液miRNAs并探索使用这些唾液miRNAs（与AD相关的SNP组合）预测AD的可能性。核心假设是长期饮酒诱导各种组织中miRNA表达的改变，并且这些改变的表达水平反映在miRNA分泌到体液如唾液中。该假设将通过追求三个具体目标来测试：（1）在非裔美国人和欧洲美国人中使用下一代测序（NGS）来描述AD受试者中唾液miRNA转录组的变化;和（2）评估差异表达的miRNA在预测AD状态中的能力;和（3）结合联合收割机AD相关的唾液miRNA和AD相关的SNP（在我们最近的全基因组关联研究中鉴定的）以改善AD的预测。我们期望发现一组AD相关的唾液miRNA和SNP，可以作为AD的生物标志物。该方法具有创新性，因为NGS技术将检查AD受试者唾液中的miRNA转录组改变，该技术在基因表达检测方面提供了前所未有的灵敏度和特异性。此外，整合miRNA表达和SNP基因型数据有望大大改善AD的预测。我们的长期目标是将鉴定的AD相关唾液miRNA用作新的诊断和预后生物标志物，并作为AD和AD相关疾病的潜在治疗靶点。这项研究具有重要意义，因为所鉴定的唾液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开发的研究策略可以应用于识别对其他物质使用障碍很重要的细胞外miRNA。

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.