Haïtham Mirghani1, Odile Casiraghi2, Joanne Guerlain3, Furrat Amen4, Ming-Xiao He5, Xiao-Jun Ma5, Yuling Luo5, Céline Mourareau6, Françoise Drusch7, Aïcha Ben Lakdhar2, Antoine Melkane3, Lacau St Guily8, Cécile Badoual9, Jean Yves Scoazec10, Isabelle Borget11, Anne Aupérin11, Veronique Dalstein6, Philippe Vielh12. 1. Department of Otolaryngology - Head and Neck Surgery, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France. Electronic address: haitham.mirghani@gustaveroussy.fr. 2. Department of Biopathology, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France. 3. Department of Otolaryngology - Head and Neck Surgery, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France. 4. Department of Otolaryngology, Peterborough City Hospital and Addenbrooke's Hospital, Cambridge, UK. 5. Advanced Cell Diagnostics, 3960 Point Eden Way, Hayward, CA 94545, USA. 6. INSERM UMR-S 903, SFR CAP-Santé FED 4231, Université de Reims Champagne-Ardenne, F-51100 Reims, France. 7. Biobank, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France. 8. Department of Otolaryngology-Head and Neck Surgery, Hôpital Tenon, Assistance Publique-Hôpitaux de Paris, France. 9. Department of Pathology, Hôpital Européen Georges Pompidou, Assistance Publique-Hôpitaux de Paris, France. 10. Department of Biopathology, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France; Biobank, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France; Laboratory of Translational Research, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France. 11. Department of Biostatistics and Epidemiology, Gustave Roussy Cancer Campus and University Paris-Sud, Villejuif, France. 12. Department of Biopathology, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France; Laboratory of Translational Research, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France.
Abstract
BACKGROUND: Accurate identification of HPV-driven oropharyngeal cancer (OPC) is a major issue and none of the current diagnostic approaches is ideal. An in situ hybridization (ISH) assay that detects high-risk HPV E6/E7 mRNA, called the RNAscope HPV-test, has been recently developed. Studies have suggested that this assay may become a standard to define HPV-status. METHODS: To further assess this test, we compared its performance against the strategies that are used in routine clinical practice: p16 immunohistochemistry (IHC) as a single test and algorithms combining p16-IHC with HPV-DNA identification by PCR (algorithm-1) or ISH (algorithm-2). RESULTS: 105 OPC specimens were analyzed. The prevalence of HPV-positive samples varied considerably: 67% for p16-IHC, 54% for algorithm-1, 61% for algorithm-2 and 59% for the RNAscope HPV-test. Discrepancies between the RNAscope HPV-test and p16-IHC, algorithm-1 and 2 were noted in respectively 13.3%, 13.1%, and 8.6%. The 4 diagnostic strategies were able to identify 2 groups with different prognosis according to HPV-status, as expected. However, the greater survival differential was observed with the RNAscope HPV-test [HR: 0.19, 95% confidence interval (CI), 0.07-0.51, p=0.001] closely followed by algorithm-1 (HR: 0.23, 95% CI, 0.08-0.66, p=0.006) and algorithm-2 (HR: 0.26, 95% CI, 0.1-0.65, p=0.004). In contrast, a weaker association was found when p16-IHC was used as a single test (HR: 0.33, 95% CI, 0.13-0.81, p=0.02). CONCLUSIONS: Our findings suggest that the RNAscope HPV-test and p16-based algorithms perform better that p16 alone to identify OPC that are truly driven by HPV-infection. The RNAscope HPV-test has the advantage of being a single test.
BACKGROUND: Accurate identification of HPV-driven oropharyngeal cancer (OPC) is a major issue and none of the current diagnostic approaches is ideal. An in situ hybridization (ISH) assay that detects high-risk HPV E6/E7 mRNA, called the RNAscope HPV-test, has been recently developed. Studies have suggested that this assay may become a standard to define HPV-status. METHODS: To further assess this test, we compared its performance against the strategies that are used in routine clinical practice: p16 immunohistochemistry (IHC) as a single test and algorithms combining p16-IHC with HPV-DNA identification by PCR (algorithm-1) or ISH (algorithm-2). RESULTS: 105 OPC specimens were analyzed. The prevalence of HPV-positive samples varied considerably: 67% for p16-IHC, 54% for algorithm-1, 61% for algorithm-2 and 59% for the RNAscope HPV-test. Discrepancies between the RNAscope HPV-test and p16-IHC, algorithm-1 and 2 were noted in respectively 13.3%, 13.1%, and 8.6%. The 4 diagnostic strategies were able to identify 2 groups with different prognosis according to HPV-status, as expected. However, the greater survival differential was observed with the RNAscope HPV-test [HR: 0.19, 95% confidence interval (CI), 0.07-0.51, p=0.001] closely followed by algorithm-1 (HR: 0.23, 95% CI, 0.08-0.66, p=0.006) and algorithm-2 (HR: 0.26, 95% CI, 0.1-0.65, p=0.004). In contrast, a weaker association was found when p16-IHC was used as a single test (HR: 0.33, 95% CI, 0.13-0.81, p=0.02). CONCLUSIONS: Our findings suggest that the RNAscope HPV-test and p16-based algorithms perform better that p16 alone to identify OPC that are truly driven by HPV-infection. The RNAscope HPV-test has the advantage of being a single test.
Keywords:
E6/E7 mRNA/ transcripts; Head and neck cancer; Human papillomavirus 16; In situ hybridization; Oropharyngeal; Oropharynx; P16 immunostaining; RNAscope HPV-test®
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