Diagnostic utility of high‐risk human papillomavirus mRNA in situ hybridisation in squamous cell carcinoma of unknown primary in the head and neck and implementing American Society of Clinical Oncology guideline recommendations

Abstract

Introduction: The American Society of Clinical Oncology (ASCO)-endorsed College of American Pathologists guideline recommends high-risk human papillomavirus (HPV) testing for metastatic squamous cell carcinoma (SCC) of lymph nodes level II/ III of unknown primary. Herein, the performance of HPV-RNA in situ hybridisation (ISH) in detection of HPV-related SCC is evaluated implementing the ASCO guideline recommendations. Methods: Eighty head and neck (HN) SCC fine needle aspirations, which utilized HPV-RNA ISH/P16, were evaluated at Johns Hopkins Hospital (2015-2018) to investigate their performance and concordance with histology. The results were compared to a prior study of 59 HNSCCs, which HPV-DNA ISH. Results: Of the 80 reviewed fine needle aspirations, 65 (50 male, 15 female) were included. The mean age was 63.2 ± 14.0 years. The most common site was neck lymph nodes (47, 72.3%). Fifty-five cases (84.6%) were accompanied by concurrent core biopsy, and 48 cases (59.4%) had surgical follow-ups. HPV-RNA ISH was positive in 44 (67.7%), and P16 was strongly positive in 46 (70.8%). The HPV-RNA ISH/ P16 concordance rate was 92.3% on cytology material. The cytology/surgical concordance rate for HPV-RNA ISH was 88.9% (16/18). There was a discordance between the results in five cases (7.7%; HPV-RNA ISH-/P16+). Conclusion: HPV-RNA ISH is a robust and reliable method for detecting HPV-related HNSCC on cytology material showing concordance rate of 92.3% between HPV-RNA ISH and P16, which is a sensitive but non-specific marker. Compared to HPV-DNA ISH, HPV-RNA ISH reproducibly identifies HPV-related HNSCC with fewer discrepancies between cytology and histology. The findings of this study are in agreement with the ASCO recommendations.

Evaluation of the Suitability of RNAscope as a Technique to Measure Gene Expression in Clinical Diagnostics: A Systematic Review

“…In contrast, Daneshpajouhnejad et al . [ 46 ] (#6) nominated RNAscope over DNA ISH for the diagnostic process as it provides an interface with easier features for interpretation. Gafeer et al .…”

Head and Neck Cystic Lesions: A Cytology Review of Common and Uncommon Entities

“…RNA probes are often complementary to E6/E7 mRNA. Since it only targets transcriptionally active viruses, it does not detect bystander viruses [36][37][38].…”

Human Papillomavirus and Squamous Cell Carcinoma of Unknown Primary in the Head and Neck Region: A Comprehensive Review on Clinical Implications

“…HPV–DNA testing can be performed by PCR with a sensitivity of 95% and specificity of 100% or in situ hybridization with a sensitivity and specificity of 91% and 94%, respectively [ 18 , 29 ], with an excellent concordance between HPV status of the cervical metastasis and identified primary tumors [ 12 ]. p16-positive (+) immunocytochemistry has similarly been proposed as a surrogate marker for HPV+ with a reasonable concordance with HPV-DNA/mRNA detection, but the technique has a notable inferior specificity [ 29 , 61 ] and is currently widely debated in terms of determining a cutoff criterion for p16 staining [ 18 ]. The technique additionally carries a risk of false positivity in benign conditions, e.g., branchial cleft cysts [ 43 , 62 ].…”