Detection of HPV infection in urothelial carcinoma using RNAscope: Clinicopathological characterization

Abstract

Background Human papillomavirus (HPV) is a well‐established mucosotropic carcinogen, but its impact on urothelial neoplasm is unclear. We aimed to clarify the clinical and pathological features of HPV‐related urothelial carcinoma (UC). Methods Tissue samples of 228 cases of UC were obtained from the bladder, upper and lower urinary tract, and metastatic sites to construct a tissue microarray. The samples were analyzed for the presence of HPV by a highly sensitive and specific mRNA in situ hybridization (RISH) technique (RNAscope) with a probe that can detect 18 varieties of high‐risk HPV. We also conducted immunohistochemistry (IHC) for a major HPV capsid antibody and DNA‐PCR. Results The HPV detection rates varied among the methods; probably due to low HPV copy numbers in UC tissues and the insufficient specificity and sensitivity of the IHC and PCR assays. The RISH method had the highest accuracy and identified HPV infection in 12 (5.2%) of the cases. The histopathological analysis of the HPV‐positive UC showed six cases of usual type UC, five cases of UC with squamous differentiation (UC_SqD), and one case of micropapillary UC. The HPV detection rate was six‐fold higher in the cases of UC_SqD than in the other variants of UC (odds ratio [OR] =8.9, p = 0.002). In addition, HPV infection showed a significant association with tumor grade (OR =9.8, p = 0.03) and stage (OR =4.7, p = 0.03) of UC. Moreover, the metastatic rate was higher in HPV‐positive than in negative UC (OR =3.4). Conclusion These data indicate that although the incidence of HPV infection in UC is low, it is significantly associated with squamous differentiation and poor prognosis. Furthermore, our observations show that RNAscope is an ideal method for HPV detection in UC compared with the other standard approaches such as IHC and PCR assays.

Prevalence of Human Papilloma Virus Infection in Bladder Cancer: A Systematic Review

“…Studies were published during the period 1995 [ 31 , 35 , 36 , 38 ]–2021 [ 44 , 57 ]. Patients were enrolled between 1985 [ 17 ] and 2019 [ 57 ].…”

“…Patients were enrolled between 1985 [ 17 ] and 2019 [ 57 ]. The epidemiological study types were observational retrospective (26, 56.5%) [ 15 , 16 , 17 , 18 , 23 , 24 , 25 , 26 , 27 , 29 , 30 , 31 , 34 , 35 , 36 , 38 , 39 , 40 , 44 , 46 , 52 , 55 , 56 , 57 , 58 , 59 ], case-control (11, 23.9%) [ 14 , 19 , 20 , 32 , 37 , 42 , 47 , 49 , 50 , 53 , 54 ], prospective (8, 17.4%) [ 21 , 22 , 33 , 41 , 43 , 45 , 48 , 51 ], and comparative (1, 2.2%) [ 28 ]. Most of them were single-center (44/46, 95.7%) [ 14 , ...…”

“…Most of them were single-center (44/46, 95.7%) [ 14 , 15 , 16 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 ], and only two (4.3%) were multi-center [ 17 , 18 ]. Single-center studies were performed in Europe (18, 40.9%) [ 27 , 28 , 29 , 30 , 35 ,…”

“…The sample size ranged from 10 [ 31 ] to 689 [ 39 ] patients, for a total of 3975 subjects. Information on gender was reported by 36 (73.5%) studies [ 14 , 15 , 16 , 17 , 19 , 20 , 21 , 22 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 37 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 50 , 51 , 53 , 55 , 56 , 57 , 58 ], including 555 and 1969 females and males, respectively. The mean/median age ranged from 47 [ 26 ] to 74.8 [ 44 ] years ( Table S2 ).…”

“…The majority of the samples were FFPE (2706/3518; 76.9%) [ 14 , 15 , 16 , 17 , 18 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 48 , 49 , 50 , 51 , 53 , 55 , 56 , 58 , 59 ], and only 812 (23.1%) were fresh tissue specimens [ 19 , 28 , 29 , 42 , 47 , 48 , 50 , 54 , 57 , 59 ]. Information on the type of specimen was not available for three (3/46; 6.5%) studies [ 27 , 39 , 52 ].…”