Screening Test for HPV+ Head and Neck Cancer

描述（由申请人提供）：目前没有好的方法来筛查由高危类型的人乳头瘤病毒引起的头颈癌的早期阶段。该提案的目的是开发一种用于牙科患者的筛查检测方法，以基于病毒和宿主基因生物标志物的使用，在术前检测HPV+头颈癌（“HNC”）。该建议的总体假设是，随着HPV相关HNC的发展，发生了许多病毒和细胞事件，可以用作生物标志物。这种生物标志物的组合为HPV转化的细胞产生了分子特征，因此可用于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，来自这些癌症的细胞可以在唾液中找到足够数量用于检测这些生物标志物的概率很高，尽管另一种可用于筛查的液体是血液。这一假设的证据已经发表。我们将开始通过在高密度阵列平台上使用新的测定（qNPA”测定）来分析与HPV+和HPV- HNC相关的基因表达变化。在这项检测中，我们将验证先前研究中涉及的基因作为推定的生物标志物，并检测编码HPV相关基因的病毒DNA和mRNA，以及涵盖细胞途径和受体的其他基因和miRNA。我们将使用组织样本和匹配的血液和唾液样本来鉴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。该提案的总体目标是开发一种快速、准确和具有成本效益的诊断工具，用于使用唾液作为样本源来早期识别头颈部区域中的癌前病变和癌性病变，其中I期SBIR是使用高密度阵列测量来识别特征基因。该计划的目标是，在后续II期研究中开发的最终诊断检测组将由参考实验室对牙科诊所收集的样本进行检测，作为常规患者评估的一部分，以识别HPV感染和HNC。第一阶段将确定用于口腔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（第一阶段，目标4）设计一个集中的诊断阵列测试（或测试），以在第二阶段资助申请中开发和验证，这也将资助对来自大量牙科患者的可接近流体（预计是唾液，但也可以是血液）样本的测试。预计该总体计划将导致针对以下各项的经验证的诊断测试：i）口服HPV; ii）HPV+ HNC; iii）HPV-HNC;以及iv）HNC的症状前识别。 公共卫生相关性：目前，还没有好的方法来筛查由人乳头瘤病毒（HPV）引起的头颈癌（“HNC”）。我们提出，随着病毒感染这些细胞，癌症开始发展，受影响细胞中表达的mRNA和miRNA发生变化，这些变化可以被检测到并用作癌症发展的“生物标志物”，而且，这些变化可以在容易获得的材料中检测到，如血液或唾液。该提案旨在开发一组可以在唾液中检测到的生物标志物，从而更容易识别那些正在发展或有发展HNC风险的个体，特别是与HPV相关的个体。我们的目标是在常规牙科就诊期间快速准确地检测早期HPV相关HNC。

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.