Circulating HPV DNA as a Prediagnostic Marker of Oropharyngeal Cancer

项目摘要 HPV驱动的口咽癌（HPV-OPC）是一种头颈部癌症，其发病率在2010年迅速增加。 美国（US）。HPV-OPC最近超过宫颈癌成为最常见的HPV相关恶性肿瘤。 与宫颈癌不同，HPV-OPC没有早期检测的方法。HPV 16 E6抗体阳性 被鉴定为HPV-OPC的有希望的早期生物标志物。本课题组的前期工作表明，HPV 16 E6抗体 存在于高达90%的HPV-OPC患者中，并且在诊断前10年以上出现。但鉴于 HPV 16 E6血清转化和癌症诊断之间的长滞后时间，HPV 16 E6血清阳性个体可能需要 在癌症可被临床诊断之前跟踪十年或更长时间。因此，早期检测生物标志物表明， 在考虑将HPV 16 E6抗体检测用于临床之前，需要确定是否存在肿瘤。循环 肿瘤（CT）HPV DNA检测是检测HPV-OPC的一种有前途的标志物。我们小组的工作证明了 ctHPVDNA对HPV-OPC诊断的敏感性为98.4%，特异性为98.6%。ctHPVDNA的既往研究 已经集中于ctHPVDNA用于诊断HPV-OPC和监测治疗后复发的效用。到 迄今为止，尚未探索ctHPVDNA用于HPV-OPC早期检测的潜力。评估的主要障碍 ctHPVDNA在诊断前环境中的应用是，迄今为止开发的检测需要大量血液（> 1 ml）， 排除前瞻性队列中的巢式研究，因为HPV-OPC诊断前采集的血液样本是 有限而珍贵。利用我们在ctDNA方面的经验，我们重新优化了基于微滴数字PCR（ddPCR）的 用于检测小体积存档标本中ctHPVDNA的测定。这项研究的目的是进行第一次 一项大型研究，旨在评估诊断前ctHPVDNA的动力学，并将该生物标志物与HPV 16 E6进行比较 血清阳性将在前列腺、肺、结直肠和卵巢（PLCO）中进行巢式病例对照研究 癌症筛查试验，一项对来自美国各地的154，935名中年男性和女性进行的前瞻性队列研究。预处理 将对来自81例OPC病例和162例匹配对照（1：2比例）的诊断血液样本进行ctHPVDNA检测。所有串行 还将测试来自ctHPVDNA阳性病例的样本，以评估导致诊断的标记物的动力学。我们 假设ctHPVDNA在癌症诊断之前是可检测，且ctHPVDNA比 HPV 16 E6血清阳性。该建议是创新的，因为使用了一种新的ctHPVDNA测定法，该测定法专门针对 小体积的存档标本，这将使我们能够进行第一项研究，以评估ctHPVDNA在一个前， 诊断设置。这项研究可能会导致更好的方法，早期检测HPV-OPC和直接回应， 由美国国家癌症研究所（NCI）和美国国家牙科和口腔医学研究所联合发布的特别关注通知 颅面研究（NIDCR）：推进头颈癌早期检测研究（AHEAD）; NOT-CA-20-031。

PROJECT SUMMARY The incidence of HPV-driven oropharyngeal cancer (HPV-OPC), a type of head and neck cancer, is rapidly increasing in the United States (US). HPV-OPC recently surpassed cervical cancer as the most common HPV-associated malignancy. Unlike cervical cancer, there are no methods of early detection for HPV-OPC. HPV16 E6 antibody positivity has been identified as a promising early biomarker of HPV-OPC. Previous work from our group showed that HPV16 E6 antibodies are present in up to 90% of HPV-OPC patients and appear more than 10 years prior to diagnosis. However, given the long lag time between HPV16 E6 seroconversion and cancer diagnosis, HPV16 E6 seropositive individuals may need to be followed for a decade or more before the cancer is clinically diagnosable. Thus, early detection biomarkers that indicate the presence or absence of tumor are needed before considering HPV16 E6 antibody testing for clinical use. Circulating tumor (ct)HPV DNA detection is a promising marker for detection of HPV-OPC. Work from our group has demonstrated that ctHPVDNA is 98.4% sensitive and 98.6% specific for HPV-OPC at the time of diagnosis. Prior studies of ctHPVDNA have focused on the utility of ctHPVDNA for diagnosing HPV-OPC and monitoring for recurrence post-treatment. To date, the potential of ctHPVDNA for early detection of HPV-OPC has not been explored. A major barrier to evaluating ctHPVDNA in a pre-diagnostic setting is that assays developed to date require a high volume of blood (>1ml), which has precluded nested studies in prospective cohorts given that blood samples collected prior to HPV-OPC diagnosis are limited and precious. Leveraging our experience in ctDNA, we have re-optimized our droplet digital PCR (ddPCR) based assay for detecting ctHPVDNA in small volumes of archived specimen. The objective of this study is to conduct the first large study to evaluate the kinetics of ctHPVDNA prior to diagnosis and to compare this biomarker to HPV16 E6 seropositivity. A nested case-control study will be conducted within the Prostate Lung Colorectal and Ovarian (PLCO) Cancer Screening Trial, a prospective cohort study of 154,935 middle aged men and women from across the US. Pre- diagnostic blood samples from 81 OPC cases and 162 matched controls (1:2 ratio) will be tested for ctHPVDNA. All serial samples from ctHPVDNA positive cases will also be tested to evaluate kinetics of the marker leading up to diagnosis. We hypothesize that ctHPVDNA will be detectable prior to cancer diagnosis and that ctHPVDNA will be more sensitive than HPV16 E6 seropositivity. This proposal is innovative given the use of a novel ctHPVDNA assay specifically optimized for small volumes of archived specimen, which will allow us to conduct the first study to evaluate ctHPVDNA in a pre- diagnostic setting. This research may lead to better methods for early detection of HPV-OPC and directly responds to the joint Notice of Special Interest issued by the National Cancer Institute (NCI) and National Institute for Dental and Craniofacial Research (NIDCR): Advancing Head and Neck Cancer Early Detection Research (AHEAD); NOT-CA-20-031.