Screening Test for HPV+ Head and Neck Cancer

描述(申请人提供)：目前还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部癌症的早期阶段。这项建议的目的是开发一种针对牙科患者的筛查方法，以基于病毒和宿主基因生物标记物的使用，在症状前检测HPV+头颈癌(HNC)。这一建议的总体假设是，随着HPV相关HNCs的发展，一些病毒和细胞事件发生，可用作生物标记物。这种生物标记物的组合为HPV转化的细胞创造了一个分子特征，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，我们认为，由于这些肿瘤的解剖位置，这些癌症的细胞可以在唾液中找到足够数量以检测这些生物标记物的可能性很高，尽管另一种可用于筛查的液体将是血液。支持这一假说的证据已经发表。我们将首先在高密度阵列平台上使用一种新的分析方法-qNPA“分析与HPV+和HPV-HNC相关的基因表达变化。在这项试验中，我们将验证以前的研究所涉及的基因作为假定的生物标志物，并测试与HPV相关的基因、覆盖细胞通路和受体的额外基因以及miRNA的病毒DNA和mRNA编码。我们将使用组织样本和匹配的血液和唾液样本来识别那些可以在唾液中找到的候选生物标记物，它们有可能区分HPV相关的HNC、与HPV无关的HNC和对照。这项建议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，以唾液为样本来源，早期识别头颈部癌前病变和癌前病变，第一阶段SBIR是使用高密度阵列测量识别签名基因。这项计划的目标是，在后续阶段开发的最终诊断测试小组将由参考实验室对牙科办公室收集的样本进行测试，作为识别HPV感染和HNC的常规患者评估的一部分。第一阶段将确定可能的DNA、mRNA和miRNA生物标志物，用于口腔HPV的诊断和HPV+HNC的早期诊断。基于这些结果，预计可以为将在第二阶段赠款申请中开发和验证的低成本阵列微板平台(第一阶段，AIM 4)设计一种有针对性的诊断阵列测试(或多项测试)，该平台还将资助对大量牙科患者的可接触液体(预计是唾液，但也可以是血液)样本的测试。预计这一整体计划将为以下方面带来有效的诊断测试：i)口服HPV；ii)HPV+HNC；iii)HPV-HNC；以及iv)HNC的症状前鉴定。 与公共卫生相关：目前，还没有很好的方法来筛查由人乳头瘤病毒(HPV)引起的头颈部癌症(HNC)。我们认为，当病毒感染这些细胞并开始发展时，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“生物标记物”，此外，这些变化可以在血液或唾液等容易接触到的物质中检测到。这项提议旨在开发一组可以在唾液中检测到的生物标记物，从而更容易识别正在发生或有患人乳头状瘤病毒风险的人，特别是那些与HPV相关的人。其目标是能够在常规牙科就诊期间快速准确地检测出个体中早期HPV相关的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.