HPV RNA in‐situ hybridization as a diagnostic aid in papillary laryngeal lesions

Abstract

Objectives In the larynx, differentiating squamous papillomas from de‐novo papillary squamous dysplasias or squamous cell carcinomas (SCC) has significant consequences for management. Overlapping clinical presentations and cytologic changes across the spectrum of papillary lesions presents diagnostic challenges for otolaryngologists and pathologists. In this study, we evaluate whether ribonucleic acid (RNA) in‐situ hybridization (ISH) for low‐risk and high‐risk human papillomavirus (HPV) can help distinguish these lesions. Methods We constructed tissue microarrays from 97 papillary laryngeal lesions, including 61 squamous papillomas, two papillomas with dysplasia, two SCCs‐ex papilloma, 14 papillary squamous dysplasias, and 18 papillary SCCs identified at the Johns Hopkins Hospital between 2000 and 2017. We performed RNA ISH using probes for low‐risk and high‐risk HPV types. Results Low‐risk HPV RNA was identified in 55 benign papillomas (90%), two papillomas with dysplasia (100%), and two SCCs‐ex papilloma (100%) but was absent in de‐novo papillary dysplasias and SCCs (0%). High‐risk HPV RNA ISH was positive only in four papillary SCC (22%). Overall, low‐risk HPV RNA ISH was 90% sensitive and 89% specific for benign papillomas with a positive predictive value of 93% and negative predictive value of 84%. In contrast, high‐risk HPV was 20% sensitive for SCC. Conclusion Low‐risk HPV RNA ISH is a useful diagnostic adjunct for distinguishing laryngeal squamous papillomas from papillary squamous dysplasia and SCC. However, it is not entirely specific for benign processes as it is also retained in papillomas with dysplasia and SCCs‐ex papilloma. Because high‐risk HPV is rare in papillary laryngeal lesions, high‐risk HPV RNA ISH has limited utility. Level of Evidence Level 4 Laryngoscope, 130:955–960, 2020

A Novel In Vivo Model of Laryngeal Papillomavirus-Associated Disease Using Mus musculus Papillomavirus

“…Laryngeal dysplasia was severe by week 2, cancers developed by week 8, and all laryngeal disease was severe dysplasia or cancer at week 12. By contrast, in RRP, which can persist for decades, moderate or greater dysplasia is found in less than 20% of cases [ 83 , 84 , 85 ] and lesions undergo malignant transformation in <1% of children and 3–6% of adults [ 86 ]. The percent of laryngeal cancers that arise from RRP is unknown, but it is likely very small.…”

Detection of Human Papillomavirus in Squamous Lesions of the Conjunctiva Using RNA and DNA In-Situ Hybridization

“…In addition, detailed review of electronic medical records was not possible for all cases in this series, as many were older pathology consultations submitted by outside physicians. While RNA in archived formalin-fixed paraffin-embedded tissue is labile and sensitive to degradation, potentially affecting the sensitivity of in-situ hybridization, we do not believe this is of concern in our study, as all in-situ hybridization assays were performed around the time of initial histologic review [63,65].…”

“…RNA in-situ hybridization permits direct visualization of intralesional transcriptionally active virus, with probes capable of detecting E6 and E7 mRNA, and provides morphologic evaluation of nuclear HPV localization within virally infected neoplastic cells [51,52,[60][61][62][63][64][65][66][67]. In addition to the reported high sensitivity and specificity, viral mRNA in-situ hybridization continues to gain popularity due to the automated staining platform, which facilitates rapid turnaround time and standardization across diagnostic laboratories and incorporation into standard slide-based workflows [60][61][62]65,[68][69][70][71]. Here, we present a case series of in-situ hybridization for detection of LR and HR HPV in conjunctival papillomas and OSSNs with the objective of describing the detection of HPV infection in these lesions, using either DNA or RNA approaches, and determining the degree to which HPV is associated with atopy.…”

Expanded Basal Compartment and Disrupted Barrier in Vocal Fold Epithelium Infected with Mouse Papillomavirus MmuPV1

“…We found that, while vocal fold infections with MmuPV1 could cause benign disease, progression to cancer was frequent in our model, whereas it is rare in human RRP [ 4 ]. Viral gene expression is found in laryngeal tissues in our MmuPV1 infection model, as seen in human RRP [ 5 , 6 , 7 ]. However, we found MmuPV1 capsid production in squamous metaplasia only in our mouse model, while low-risk HPV capsid protein is found in sparse, superficial cells in human RRP lesions but is not absent [ 8 ].…”