Regulation of mRNA Stability in Human Saliva

描述(申请人提供)：研究正常唾液和癌症唾液中mRNA的表达和调控，为发现口腔癌的诊断标记物提供了巨大的潜力。毫无疑问，唾液是最无创的体液，可以很容易地收集和保存。先前的研究表明，唾液中含有可用于口腔癌诊断的生物标志物的人类mRNAs。David Wong博士在加州大学洛杉矶分校牙科学院的研究小组最近开发了口腔癌患者唾液中的诊断RNA标记物。这项技术是基于微阵列的，非常适合口腔病理的实时医疗诊断。这项建议的目的是为了了解人类唾液中信使核糖核酸的稳定和去稳定的作用。我们通过基因芯片鉴定的大多数口腔癌基因在其3‘非编码区序列上都含有富含AU的元件(ARE)。ARES通过其反式作用蛋白靶向mRNAs的快速降解。因此，这项建议的主要目的是确定唾液中的mRNA结合蛋白，以及什么因素参与了这一过程。在癌症条件下，含有转录物的唾液中转录产物上调并稳定。因此，我们的假设是，通过避免mRNA衰退机制，转录产物在癌症条件下是稳定的。我们计划利用信使核糖核酸衰变系统，结合我们之前对信使核糖核酸降解途径的研究，进一步测试、验证和确定唾液中信使核糖核酸稳定性的机制。这项建议的最初指导阶段包括以下目标：i)阐明与人唾液中mRNA稳定有关的因素。独立研究阶段包括以下目标：ii)确定癌症患者唾液中mRNAs稳定的基本机制，iii)验证ARE中含有mRNAs的衰退与MAP激酶/通路激活的相关性。综上所述，这些研究将打开正常和口腔癌唾液中RNA的稳定性，从而促进RNA生物标记物在癌症临床诊断中的进步。

DESCRIPTION (provided by applicant): Studying expression and regulation of mRNA in normal versus cancer saliva is garnering great potential for the discovery of diagnostic marker for oral cancer. Undoubtedly, saliva is the most non-invasive body fluid that can be easily collected and preserved. Previous research has shown that saliva contains human mRNAs that can be used for diagnostic biomarkers for oral cancer. Dr. David Wong's research group at the University of California Los Angeles, School of Dentistry, has recently developed diagnostic RNA markers in oral cancer patient saliva. This technology is a microarray based and well suited for real-time medical diagnostics for oral pathology. The goal of this proposal is directed toward understanding the role of mRNA stability and de-stability in human saliva. The majority of oral cancer genes which we identified through microarray contain AU-rich elements (AREs) at their 3' UTR sequences. AREs target mRNAs for rapid degradation via its trans-acting proteins. Hence the main aim of this proposal is to identify mRNA binding proteins in saliva and what factors are involved in this process. Under cancer conditions ARE containing transcripts are up-regulated and stabilized in saliva. Thus, our hypothesis is that ARE transcripts are stabilized in cancer conditions by means of avoiding mRNA decay machinery. We plan to utilize mRNA decay system along with expertise gained from our previous studies on mRNA degradation pathways so far to further test, validate and identify the mechanistic aspects of mRNA stability in saliva. The initial mentored phase of this .proposal includes the following aim: I) Elucidate the factors involved in mRNA stability in human saliva. The independent investigator phase includes the following aims: II) Determine the fundamental mechanism responsible for the stabilization of mRNA's in saliva of cancer patients, III) Validate the ARE containing mRNA decay in correlation with MAP kinase/pathway activation. In summary, these studies will open up the mRNA stability in normal and oral cancer saliva lead to advances in clinical diagnostics of RNA biomarkers in cancer.