Regulation of mRNA Stability in Human Saliva

描述（申请人提供）：研究正常与癌症唾液中mRNA的表达和调控，对于发现口腔癌的诊断标志物具有很大的潜力。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，唾液中含有人类mrna，可用于口腔癌的诊断生物标志物。加州大学洛杉矶分校牙科学院的David Wong博士的研究小组最近在口腔癌患者的唾液中开发出了诊断性RNA标记。该技术是基于微阵列的，非常适合于口腔病理的实时医学诊断。这一建议的目标是直接了解mRNA的稳定性和去稳定性在人类唾液中的作用。我们通过微阵列技术鉴定的口腔癌基因在其3' UTR序列上含有富au元素（AREs）。AREs通过其反式作用蛋白靶向mrna进行快速降解。因此，本研究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，含有转录本的ARE在唾液中被上调和稳定。因此，我们的假设是，通过避免mRNA衰变机制，ARE转录本在癌症条件下稳定下来。我们计划利用mRNA衰变系统以及迄今为止我们从mRNA降解途径的先前研究中获得的专业知识来进一步测试，验证和确定mRNA在唾液中稳定性的机制方面。最初的指导阶段。建议包括以下目的：1)阐明参与人唾液mRNA稳定性的因素。独立研究者阶段包括以下目标：II)确定癌症患者唾液中mRNA稳定的基本机制，III)验证含有mRNA衰减与MAP激酶/通路激活的相关性。综上所述，这些研究将揭示正常和口腔癌唾液中mRNA的稳定性，从而推动RNA生物标志物在癌症中的临床诊断。

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.