Abstract

OBJECTIVES: In this feasibility study, we explored the combined use of circulating tumor human papillomavirus (HPV) DNA (ctHPVDNA) and HPV serology as diagnostic tests for HPV-associated oropharyngeal squamous cell carcinoma (OPSCC).

METHODS: Among patients with research-banked serum or plasma at diagnosis, IgG antibodies to oncoproteins from HPV types 16, 18, 31, 33, 35, 45, 52, and 58 were detected with multiplex serology. Positivity for HPV 16 was defined based on detection of combinations of anti-E6, E1, E2, and E7 and for other high-risk types on detection of anti-E6 and anti-E7. Circulating tumor HPV DNA was detected by custom digital droplet polymerase chain reaction (ddPCR) assays for HPV types 16, 18, 33, 35, and 45. p16 immunohistochemistry and high-risk HPV RNA in situ hybridization (ISH) using a cocktail of 18 high-risk HPV types were performed on tissue.

RESULTS: Of 75 patients, 67 (89.3%) were HPV-associated (p16 and HPV RNA ISH positive) and 8 (10.7%) were HPV-independent. All 8 HPV-independent patients were seronegative and negative for ctHPVDNA (100% specificity). Serology was positive in 53 (79.1%) of 67 HPV-associated patients, while ddPCR was positive for ctHPVDNA in 59 (88.6%) of 67 HPV-associated patients. Requiring both tests to be positive resulted in a sensitivity of 50 (74.6%) of 67 while combining assays (either positive) improved sensitivity to 62 (92.6%) of 67.

CONCLUSIONS: Compared to HPV RNA ISH, HPV serology and ctHPVDNA are sensitive and highly specific biomarkers for HPV-associated OPSCC at the time of presentation.

Document Type

Article

Publication Date

6-3-2024

Notes/Citation Information

© The Author(s) 2024. Published by Oxford University Press on behalf of American Society for Clinical Pathology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.

Digital Object Identifier (DOI)

https://doi.org/10.1093/ajcp/aqad185

Funding Information

Daniel L. Faden receives salary support from National Institutes of Health grants (K23DE029811, R03DE030550, NIH R21CA267152) and also receives support from Calico; holds equity in Illumina; receives consulting fees from Merck, Noetic, Chrysalis Biomedical Advisors, and Focus; and receives research funding from BMS, Predicine, and BostonGene. Funding for this work was supported by National Institutes of Health under award number R03DE030550 and also supported by the National Cancer Institute of the National Institutes of Health under award number K07CA218247 (principal investigator: Krystle A. Lang Kuhs). Tim Waterboer serves on the advisory boards of Merck and Sharp and Dohme (MSD). James S. Lewis Jr receives salary support from National Institutes of Health grants (1R01CA249992-01A1, 1R01CA220581-01A1) and Department of Defense grant (Award # W81XWH2110160 Subaward # RES516237). The Vanderbilt Tissue Pathology Shared Resource, which constructed the tissue microarrays and performed the p16 immunohistochemistry and high-risk HPV RNA ISH for this study, is supported by the National Cancer Institute of the National Institutes of Health Cancer Center Support Grant (P30CA068485), the Shared Instrumentation Grant (S10 OD023475-01A1) for the Leica Bond RX, and the Shared Instrumentation Grant (S10 OD016355) for the TMA Grandmaster. The other authors have nothing to disclose.

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