Damerla Rama R, Lee Nancy Y, You Daoqui, Soni Rekha, Shah Rachna, Reyngold Marsha, Katabi Nora, Wu Vanessa, McBride Sean M, Tsai Chiaojung Jillian, Riaz Nadeem, Powell Simon N, Babady N Esther, Viale Agnes, Higginson Daniel S
Memorial Sloan-Kettering Cancer Center, New York, NY.
JCO Precis Oncol. 2019;3. doi: 10.1200/PO.18.00276. Epub 2019 Apr 3.
A circulating tumor DNA (ctDNA) test to detect plasma Epstein-Barr viral DNA can be used to screen for early nasopharyngeal cancers; however, the reported sensitivity of viral ctDNA tests to detect human papillomavirus (HPV)-associated cancers is modest. We assessed the utility of droplet digital polymerase chain reaction (ddPCR) to detect early-stage HPV-associated cancers using sequential HPV16 and HPV33 assays that account for HPV subtype distribution and subtype sequence variants.
We collected plasma specimens from 97 HPV-positive patients with oropharyngeal squamous cell carcinoma and eight patients with HPV-positive anal squamous cell carcinoma, each with locoregionally confined disease. Negative controls included samples from seven patients with HPV-negative head and neck cancers and 20 individuals without cancer.
Of 97 patients with nonmetastatic, locoregionally confined oropharyngeal squamous cell carcinoma, 90 patients had detectable HPV16 ctDNA and three patients had HPV33 ctDNA, indicating an overall sensitivity of 95.6%. Seven of eight patients with early anal cancer were HPV16 ctDNA positive. No HPV ctDNA was detected in 27 negative controls, indicating 100% specificity. HPV16 ctDNA was detected in 19 of 19 patients with low-volume disease, defined as patients with a single, asymptomatic positive lymph node (N1) or an isolated T1-2 asymptomatic primary tumor. HPV16 ctDNA levels directly corresponded to tumor responses to chemoradiation and surgery.
With an updated understanding of HPV subtypes and sequence variation, HPV ctDNA by ddPCR is highly sensitive and specific, identifying HPV16 and HPV33 subtypes in a similar distribution as reported in major genomic profiling studies. The detection of small tumors indicates that HPV16 and HPV33 ctDNA ddPCR could be readily used in early detection screening trials and in disease response monitoring, analogous to Epstein-Barr virus DNA.
一种用于检测血浆中爱泼斯坦-巴尔病毒DNA的循环肿瘤DNA（ctDNA）检测方法可用于早期鼻咽癌的筛查；然而，报告显示病毒ctDNA检测对人乳头瘤病毒（HPV）相关癌症的敏感性一般。我们使用考虑了HPV亚型分布和亚型序列变异的HPV16和HPV33序列检测方法，评估了液滴数字聚合酶链反应（ddPCR）检测早期HPV相关癌症的效用。
我们收集了97例HPV阳性的口咽鳞状细胞癌患者以及8例HPV阳性的肛门鳞状细胞癌患者的血浆样本，这些患者均患有局限性疾病。阴性对照包括7例HPV阴性的头颈癌患者和20例无癌症个体的样本。
在97例非转移性、局限性口咽鳞状细胞癌患者中，90例患者可检测到HPV16 ctDNA，3例患者可检测到HPV33 ctDNA，总体敏感性为95.6%。8例早期肛门癌患者中有7例HPV16 ctDNA呈阳性。27例阴性对照中未检测到HPV ctDNA，特异性为100%。在19例疾病负荷低的患者（定义为有单个无症状阳性淋巴结（N1）或孤立的无症状T1-2期原发肿瘤的患者）中，有19例检测到HPV16 ctDNA。HPV16 ctDNA水平与肿瘤对放化疗和手术的反应直接相关。
随着对HPV亚型和序列变异的进一步了解，通过ddPCR检测HPV ctDNA具有高度敏感性和特异性，可识别出与主要基因组分析研究报告的分布相似的HPV16和HPV33亚型。对小肿瘤的检测表明，HPV16和HPV33 ctDNA ddPCR可很容易地用于早期检测筛查试验和疾病反应监测，类似于爱泼斯坦-巴尔病毒DNA。
