Salivary MicroRNAs as Biomarkers for Alcohol Dependence

描述（由申请人提供）：酒精依赖（AD）是一种严重的公共健康问题，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。识别有效的AD生物标志物对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，在转录后水平调控靶基因表达的小非编码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（即唾液、血清、尿液等）中存在mirna，这表明细胞外mirna是疾病诊断和预后的潜在生物标志物。迄今为止，关于人类阿尔茨海默病患者miRNA表达变化的信息非常有限，目前还没有研究检测阿尔茨海默病患者唾液中miRNA表达的变化。本应用程序的目的是鉴定AD相关的唾液mirna，并探索使用这些唾液mirna（与AD相关的snp结合）预测AD的可能性。核心假设是，长期饮酒会导致各种组织中miRNA表达的改变，而这些改变的表达水平反映在miRNA分泌到唾液等体液中。这一假设将通过追求三个具体目标来验证：(1)使用下一代测序（NGS）在非洲裔美国人和欧洲裔美国人中分析AD受试者的唾液miRNA转录组变化；(2)评估差异表达的mirna预测AD状态的能力；(3)结合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发现）来改善AD的预测。我们希望发现一组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，因为将通过NGS技术检测AD受试者唾液中的miRNA转录组变化，该技术在检测基因表达方面具有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提高对AD的预测。我们的长期目标是利用已鉴定的AD相关唾液mirna作为新的诊断和预后生物标志物，以及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mirna有望在监测AD的发展方面具有临床意义。此外，本研究提出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.