Regulation of mRNA Stability in Human Saliva

描述（由申请人提供）：研究mRNA在正常唾液与癌唾液中的表达和调节对于发现口腔癌的诊断标志物具有巨大的潜力。毫无疑问，唾液是最容易收集和保存的非侵入性体液。先前的研究表明，唾液中含有人类mRNA，可用于口腔癌的诊断生物标志物。洛杉矶加州牙科学院的大卫王博士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。该技术是基于微阵列的，非常适合用于口腔病理学的实时医学诊断。该提案的目标是了解人类唾液中mRNA稳定性和去稳定性的作用。我们通过微阵列鉴定的大多数口腔癌基因在其3' UTR序列处含有富含AU的元件（战神）。战神靶向mRNA，通过其反式作用蛋白快速降解。因此，该提案的主要目的是确定唾液中的mRNA结合蛋白以及参与这一过程的因素。在癌症条件下，含有ARE的转录物在唾液中上调并稳定。因此，我们的假设是ARE转录物通过避免mRNA衰变机制在癌症条件下稳定。我们计划利用mRNA衰变系统沿着，结合我们之前对mRNA降解途径的研究所获得的专业知识，进一步测试、验证和鉴定唾液中mRNA稳定性的机制方面。该建议的初始指导阶段包括以下目标：I）阐明涉及人唾液中mRNA稳定性的因素。独立研究者阶段包括以下目标：II）确定负责癌症患者唾液中mRNA稳定的基本机制，III）确定与MAP激酶/途径活化相关的含有ARE的mRNA衰变。总之，这些研究将打开正常和口腔癌唾液中mRNA的稳定性，从而推动癌症RNA生物标志物的临床诊断。

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.