Lisa M Rooper1, Manoj Gandhi2, Justin A Bishop3, William H Westra4. 1. Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD, USA. 2. Affymetrix, Inc., Santa Clara, CA, USA. 3. Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD, USA; Department of Otolaryngology-Head and Neck Surgery, The Johns Hopkins Medical Institutions, Baltimore, MD, USA. 4. Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD, USA; Department of Otolaryngology-Head and Neck Surgery, The Johns Hopkins Medical Institutions, Baltimore, MD, USA; Department of Oncology, The Johns Hopkins Medical Institutions, Baltimore, MD, USA. Electronic address: wwestra@jhmi.edu.
Abstract
OBJECTIVES: Evaluation of human papillomavirus (HPV) status in oropharyngeal squamous cell carcinoma (OPSCC) has become increasingly important for prognostication and clinical trial enrollment. This assessment is confounded in OPSCCs that are p16 positive by immunohistochemistry (IHC) but HPV negative by DNA in situ hybridization (DISH). This study evaluates whether E6/E7 mRNA in situ hybridization (RISH) can detect transcriptionally active HPV in these problematic cases. MATERIALS AND METHODS: Eighty-two head and neck squamous cell carcinoma cases that had previously undergone p16 IHC and HPV DISH were evaluated with two RISH platforms and a second-generation DISH probe. The study included 21 p16+/DISH+ concordant cases, 19 p16-/DISH- concordant cases, and 42 p16+/DISH- discordant cases. RESULTS: RISH identified E6/E7 mRNA in 37 (88%) p16+/DISH- cases, 21 (100%) p16+/DISH+ cases, and 0 (0%) p16-/DISH- cases. RISH signals were clearly visible at low to medium magnification in 97% of positive cases, facilitating almost-perfect inter-observer reproducibility. The performance of the manual and automated RISH platforms were equivalent (kappa=0.915). Only 29% of carcinomas that demonstrated E6/E7 mRNA transcriptional activity were positive using the 2nd generation DISH probe. CONCLUSIONS: HPV RISH is a highly sensitive and specific platform that can clarify the HPV status of those perplexing OPSCCs that are p16 positive by IHC but HPV negative by DISH. Moreover, it is easy to interpret, readily adaptable to the clinical laboratory, and provides direct evidence of HPV transcriptional activity. E6/E7 RISH should be considered as a first-line platform for determination of HPV status in OPSCCs.
OBJECTIVES: Evaluation of human papillomavirus (HPV) status in oropharyngeal squamous cell carcinoma (OPSCC) has become increasingly important for prognostication and clinical trial enrollment. This assessment is confounded in OPSCCs that are p16 positive by immunohistochemistry (IHC) but HPV negative by DNA in situ hybridization (DISH). This study evaluates whether E6/E7 mRNA in situ hybridization (RISH) can detect transcriptionally active HPV in these problematic cases. MATERIALS AND METHODS: Eighty-two head and neck squamous cell carcinoma cases that had previously undergone p16 IHC and HPV DISH were evaluated with two RISH platforms and a second-generation DISH probe. The study included 21 p16+/DISH+ concordant cases, 19 p16-/DISH- concordant cases, and 42 p16+/DISH- discordant cases. RESULTS: RISH identified E6/E7 mRNA in 37 (88%) p16+/DISH- cases, 21 (100%) p16+/DISH+ cases, and 0 (0%) p16-/DISH- cases. RISH signals were clearly visible at low to medium magnification in 97% of positive cases, facilitating almost-perfect inter-observer reproducibility. The performance of the manual and automated RISH platforms were equivalent (kappa=0.915). Only 29% of carcinomas that demonstrated E6/E7 mRNA transcriptional activity were positive using the 2nd generation DISH probe. CONCLUSIONS:HPV RISH is a highly sensitive and specific platform that can clarify the HPV status of those perplexing OPSCCs that are p16 positive by IHC but HPV negative by DISH. Moreover, it is easy to interpret, readily adaptable to the clinical laboratory, and provides direct evidence of HPV transcriptional activity. E6/E7 RISH should be considered as a first-line platform for determination of HPV status in OPSCCs.
Authors: Elaine S Keung; Rhona J Souers; Julia A Bridge; William C Faquin; Rondell P Graham; Meera R Hameed; James S Lewis; Jason D Merker; Patricia Vasalos; Joel T Moncur Journal: Arch Pathol Lab Med Date: 2019-09-04 Impact factor: 5.534
Authors: Lisa M Rooper; Melina J Windon; Tahyna Hernandez; Brett Miles; Patrick K Ha; William R Ryan; Annemieke Van Zante; David W Eisele; Gypsyamber D'Souza; Carole Fakhry; William H Westra Journal: Am J Surg Pathol Date: 2020-05 Impact factor: 6.394
Authors: Carole Fakhry; William H Westra; Steven J Wang; Annemieke van Zante; Yuehan Zhang; Eleni Rettig; Linda X Yin; William R Ryan; Patrick K Ha; Alicia Wentz; Wayne Koch; Jeremy D Richmon; David W Eisele; Gypsyamber D'Souza Journal: Cancer Date: 2017-02-27 Impact factor: 6.860
Authors: Peter P Luk; Christina I Selinger; Wendy A Cooper; Annabelle Mahar; Carsten E Palme; Sandra A O'Toole; Jonathan R Clark; Ruta Gupta Journal: Head Neck Pathol Date: 2018-11-22
Authors: Melina J Windon; Gypsyamber D'Souza; Eleni M Rettig; William H Westra; Annemieke van Zante; Steven J Wang; William R Ryan; Wojciech K Mydlarz; Patrick K Ha; Brett A Miles; Wayne Koch; Christine Gourin; David W Eisele; Carole Fakhry Journal: Cancer Date: 2018-04-30 Impact factor: 6.860
Authors: Linda X Yin; Gypsyamber D'Souza; William H Westra; Steven J Wang; Annemieke van Zante; Yuehan Zhang; Eleni M Rettig; William R Ryan; Patrick K Ha; Alicia Wentz; Wayne Koch; David W Eisele; Carole Fakhry Journal: Laryngoscope Date: 2018-03-14 Impact factor: 3.325