Screening Test for HPV+ Head and Neck Cancer

描述（由申请人提供）：目前没有很好的方法来筛查由高危类型的人乳头瘤病毒引起的早期头颈部癌症。本提案的目的是开发一种基于病毒和宿主基因生物标志物的筛查方法，用于牙科患者在症状前检测HPV+头颈癌（“HNC”）。该提案的总体假设是，随着hpv相关HNCs的发展，许多病毒和细胞事件的发生可以被用作生物标志物。这种生物标志物的组合为HPV转化的细胞创造了分子特征，因此可以用来识别可能患癌症的个体。更广泛地说，与HPV感染无关的HNC也是如此。此外，我们提出，由于这些肿瘤的解剖位置，在唾液中发现足够数量的这些癌症细胞以检测这些生物标志物的可能性很高，尽管另一种可用于筛选的液体是血液。这一假设的证据已经发表。我们将首先分析与HPV+和HPV- HNC相关的基因表达变化，使用高密度阵列平台上的新检测，qNPA”检测。在这个实验中，我们将验证先前研究中涉及的基因作为假定的生物标志物，并测试病毒DNA和mRNA编码与HPV相关的基因，覆盖细胞通路和受体的其他基因，以及miRNA。我们将使用组织样本和匹配的血液和唾液样本来确定那些候选生物标志物，这些生物标志物可以在唾液中找到（理想情况下），并且有可能区分HPV相关的HNC，与HPV无关的HNC和对照组。本提案的总体目标是开发一种快速、准确和经济有效的诊断工具，用于头颈部癌前病变和癌性病变的早期识别，以唾液为样本来源，第一阶段的SBIR是使用高密度阵列测量来识别特征基因。该项目的目标是，在后续二期研究中开发的最终诊断测试小组将由参考实验室对牙科诊所收集的样本进行测试，作为常规患者评估的一部分，以确定HPV感染和HNC。I期将确定用于口腔HPV诊断和HPV+ HNC早期诊断的推定DNA、mRNA和miRNA生物标志物。预计，基于这些结果，可以为低成本的ArrayPlate微孔板平台（I期，Aim 4）设计一个集中诊断阵列测试（或多个测试），并在II期拨款申请中开发和验证，这也将资助对大量牙科患者可获得的液体样本（预计是唾液，但也有可能是血液）的测试。预计这一总体规划将导致有效的诊断测试：i)口腔HPV；ii) HPV+ HNC；iii) HPV- HNC；iv) HNC的症状前识别。

DESCRIPTION (provided by applicant): There is currently no good way to screen for early stages of head and neck cancers caused by high risk types of human papillomavirus. The objective of this proposal is to develop a screening assay for dental patients to detect HPV+ head and neck cancer ("HNC") pre-symptomatically, based on the use of viral and host gene biomarkers. The overall hypothesis of this proposal is that as the development of HPV-associated HNCs proceeds, a number of viral and cellular events occur that can be exploited as biomarkers. This combination of biomarkers creates a molecular signature for cells transformed by HPV and consequently could be used to identify individuals likely to develop cancer. More broadly, the same is true of HNC not associated with HPV infection. Furthermore, we propose that, due to the anatomical location of these tumors, the probability that cells from these cancers can be found in saliva in sufficient numbers for the detection of these biomarkers is high, though another accessible fluid for screening would be blood. Evidence for this hypothesis has been published. We will begin by profiling the gene expression changes associated with HPV+ and HPV- HNC using a new assay on a high density array platform, the qNPA" assay. In this assay, we will validate genes implicated by previous studies as putative biomarkers, and also test viral DNA and mRNA coding for genes associated with HPV, additional genes that cover cellular pathways and receptors, and miRNA. We will use tissue samples and matched blood and saliva samples to identify those candidate biomarkers that can be found (ideally) in saliva and that have the potential to distinguish between HPV-associated HNC, HNC not associated with HPV, and controls. The overall goal of this proposal is to develop a rapid, accurate and cost- effective diagnostic tool for the early identification of pre-cancerous and cancerous lesions in the head and neck area, using saliva as a sample source, with Phase I SBIR being the identification of signature genes using high-density array measurements. It is the objective of this program that the ultimate diagnostic test panel developed in a follow-on Phase II will be performed by reference laboratories on samples collected in dental offices as part of routine patient assessments to identify HPV infection and HNC. Phase I will identify putative DNA, mRNA and miRNA biomarkers for use in the diagnosis of oral HPV and the early diagnosis of HPV+ HNC. It is anticipated that, based on these results, a focused diagnostic array test (or tests) can be designed for the low-cost ArrayPlate microplate platform (Phase I, Aim 4) to be developed and validated in a Phase II grant application, which will also fund the testing of samples of an accessible fluid (expected to be saliva, but alternatively blood) from a large number of dental patients. It is anticipated that this overall program will result in validated diagnostic tests for: i) oral HPV; ii) HPV+ HNC; iii) HPV- HNC; and for iv) the pre- symptomatic identification of HNC. PUBLIC HEALTH RELEVANCE: At this time, there is no good way to screen for head and neck cancers ("HNC") that are caused by Human Papillomaviruses (HPV). We propose that as the virus infects these cells and cancer begins to develop, changes in the mRNA and miRNA expressed in the affected cells occur that can be detected and used as "biomarkers" for the development of cancer, and further, that these can be detected in easily-accessible material, such as blood or saliva. This proposal is designed to develop a panel of biomarkers that could be detected in saliva, thus making it much easier to identify those individuals who are developing or are at risk for developing HNCs, especially those associated with HPV. The goal is to make it possible to quickly and accurately detect early stage HPV-associated HNC in individuals during routine dental visits.