Mena Marisa, Wang Xin, Tous Sara, Quiros Beatriz, Clavero Omar, Alejo Maria, Morey Francisca, Taberna Miren, Leon Vintro Xavier, Lloveras Rubio Belén, Alos Llúcia, Mehanna Hisham, Quint Wim, Pawlita Michael, Tommasino Massimo, Pavón Miguel Angel, Muñoz Nubia, De Sanjose Silvia, Bosch Francesc Xavier, Alemany Laia
Cancer Epidemiology Research Program, Catalan Institute of Oncology (ICO)-IDIBELL, L'Hospitalet de Llobregat, 08908 Barcelona, Spain.
Centro de Investigación Biomédica en Red de Epidemiología y Salud Pública (CIBERESP), Instituto de Salud Carlos III, 28029 Madrid, Spain.
Cancers (Basel). 2022 Aug 4;14(15):3787. doi: 10.3390/cancers14153787.
Background: Tests or test algorithms for diagnosing HPV-driven oral cavity and laryngeal head and neck carcinomas (HNC) have not been yet validated, and the differences among oral cavity and laryngeal sites have not been comprehensively evaluated. We aimed to assess the utility of a diagnostic algorithm for the detection of HPV-driven oral cavity (OCC), oropharyngeal (OPC) and laryngeal (LC) carcinomas using HPV-DNA testing followed by p16INK4a immunohistochemistry, taking E6I mRNA detection as the reference standard. Methods: Formalin-fixed paraffin-embedded OCC, OPC, and LC carcinomas were collected from pathology archives in 29 countries. All samples were subjected to histopathological evaluation, DNA quality control, and HPV-DNA detection. All HPV-DNA-positive samples (including 78 OCC, 257 OPC, and 51 LC out of 3680 HNC with valid HPV-DNA results) were also tested for p16INK4a immunohistochemistry and E6I mRNA. Three different cutoffs of nuclear and cytoplasmic staining were evaluated for p16INK4a: (a) >25%, (b) >50%, and (c) ≥70%. The concordance of p16INK4a and E6I mRNA among HPV-DNA-positive OCC, OPC, and LC cases was assessed. Results: A total of 78 OCC, 257 OPC, and 51 LC were HPV-DNA-positive and further tested for p16INK4a and E6I mRNA. The percentage of concordance between p16INK4a (cutoff ≥ 70%) and E6I mRNA among HPV-DNA-positive OCC, OPC, and LC cases was 79.5% (95% CI 69.9−89.1%), 82.1% (95% CI 77.2−87.0%), and 56.9% (95% CI 42.3−71.4%), respectively. A p16INK4a cutoff of >50% improved the concordance although the improvement was not statistically significant. For most anatomical locations and p16INK4a cutoffs, the percentage of discordant cases was higher for HPV16- than HPV-non16-positive cases. Conclusions: The diagnostic algorithm of HPV-DNA testing followed by p16INK4a immunohistochemistry might be helpful in the diagnosis of HPV-driven OCC and OPC, but not LC. A different p16INK4a expression pattern was observed in those cases HPV-DNA-positive for types other than HPV16, as compared to HPV16-positive cases. Our study provides new insights into the use HPV-DNA, p16INK4a, and HPV-E6I mRNA for diagnosing an HPV-driven HNC, including the optimal HPV test or p16INK4a cutoffs to be used. More studies are warranted to clarify the role of p16INK4a and HPV status in both OPC and non-OPC HNC.
用于诊断人乳头瘤病毒（HPV）驱动的口腔和喉头颈癌（HNC）的检测方法或检测算法尚未得到验证，口腔和喉部位之间的差异也未得到全面评估。我们旨在评估一种诊断算法在检测HPV驱动的口腔癌（OCC）、口咽癌（OPC）和喉癌（LC）中的效用，该算法采用HPV-DNA检测，随后进行p16INK4a免疫组织化学检测，并将E6I mRNA检测作为参考标准。方法：从29个国家的病理档案中收集福尔马林固定石蜡包埋的OCC、OPC和LC癌组织。所有样本均进行组织病理学评估、DNA质量控制和HPV-DNA检测。所有HPV-DNA阳性样本（在3680例有有效HPV-DNA结果的HNC中，包括78例OCC、257例OPC和51例LC）也进行了p16INK4a免疫组织化学和E6I mRNA检测。对p16INK4a的三种不同核染色和胞质染色临界值进行了评估：（a）>25%，（b）>50%，（c）≥70%。评估了HPV-DNA阳性的OCC、OPC和LC病例中p16INK4a与E6I mRNA的一致性。结果：共有78例OCC、257例OPC和51例LC为HPV-DNA阳性，并进一步进行了p16INK4a和E6I mRNA检测。在HPV-DNA阳性的OCC、OPC和LC病例中，p16INK4a（临界值≥70%）与E6I mRNA之间的一致性百分比分别为79.5%（95%CI为69.9−89.1%）、82.1%（95%CI为77.2−87.0%）和56.9%（95%CI为42.3−71.4%）。p16INK4a临界值>50%可提高一致性，尽管这种提高在统计学上不显著。对于大多数解剖部位和p16INK4a临界值，HPV16阳性病例的不一致病例百分比高于HPV非16阳性病例。结论：HPV-DNA检测随后进行p16INK-a免疫组织化学的诊断算法可能有助于诊断HPV驱动的OCC和OPC，但对LC诊断无帮助。与HPV16阳性病例相比，在HPV-DNA阳性的非HPV16型病例中观察到不同的p16INK4a表达模式。我们的研究为使用HPV-DNA、p16INK4a和HPV-E6I mRNA诊断HPV驱动的HNC提供了新的见解，包括最佳的HPV检测或p16INK4a临界值。需要更多的研究来阐明p16INK4a和HPV状态在OPC和非OPC HNC中的作用。
