Clinical and molecular analysis of JCPyV and BKPyV infection and associated risk of urothelial carcinoma development in the upper tract

Abstract

Background: Human polyomaviruses (HPyVs), JC polyomavirus (JCPyV) and BK polyomavirus (BKPyV), have been found in upper tract urothelial carcinoma UTUC; however, the association of the viral oncogenic factors and clinical characteristics of UTUC remains unclear. This study aimed to investigate the prevalence of JCPyV and BKPyV in UTUC and their correlation with cancer progression among the southwest Taiwanese population from 2020 to 2022.

Methods: A total of 72 paraffin-embedded UTUC tissue samples and 41 adjacent tissue samples were collected from 72 patients. Nested polymerase chain reaction and DNA sequencing were used to detect viral DNA and genotypes. Immunohistochemistry was performed using anti- large T (LT) and anti-p53 monoclonal antibodies to detect the expression of viral early LT protein and cellular p53 protein, respectively.

Results: The overall prevalence of JCPyV and BKPyV were higher in UTUC than in adjacent tissue samples (65.3% [47/72] vs. 17.1% [7/41]). JCPyV and BKPyV were detected in 95.7% (45/47) and 4.3% (2/47) of the HPyVs-positive UTUC samples, respectively. JCPyV-TW-3 was the predominant strain of JCPyV infection. In UTUC samples, the LT protein of JCPyV and BKPyV positivity rate was 65.3%, while that of mutant p53 protein was 52.7%. JCPyV infection and LT protein expression increased the odds ratio (OR) of UTUC by 9.13-fold. The OR of UTUC was higher by 10.34-fold in patients with mutant p53 and by 10.37-fold in those with simultaneous LT and mutant p53 expression. The presence of LT protein in UTUC patients may increase the OR of mutant p53 protein expression by 2.93-fold compared to its absence. Women had a 5.19-fold higher superiority of JCPyV infection and LT expression than men. Patients with chronic kidney disease (CKD) had a 3.15-fold higher OR for mutant p53 protein expression than those without it. In the UTUC advanced stages, the OR of virus and LT expression was 3.18-fold higher compared to those who do not require chemotherapy.

Conclusions: JCPyV infection is highly prevalent in UTUC, and the presence of CKD concurrent with high expressions of LT and mutant p53 proteins in patients may be a useful indicator for chemotherapy and poor prognosis.

Keywords: Chronic kidney disease (CKD); JCPyV infection; LT protein; Upper tract urothelial carcinoma; p53 protein.

Fig. 1

Gel electrophoresis analysis of PCR-amplified products from UTUC and adjacent tissue samples. A set of conserved primers targeting the regulatory regions of JCPyV and BKPyV is used for amplification, followed by gel analysis on a 2.5% agarose gel. Lane M contains DNA markers with molecular weights of 50, 100, 150, 200,250, 350, and 450 bp. (a) Lane numbers correspond to tissue sample numbers No. 1–72 from UTUC samples. (b) Lane numbers represent tissue sample numbers No. 1–41 from adjacent tissue samples. A positive control (P) is included using JCPyV CY and BKPyV UT genomic DNA templates, resulting in bands of approximately 243 bp and 289 bp in size, respectively. Negative control (N) indicates that PCR is performed without the DNA template. Additionally, β-actin is included as an internal control for PCR analysis. BKPyV, BK polyomavirus; JCPyV, JC polyomavirus; PCR, polymerase chain reaction; UTUC, upper tract urothelial carcinoma

Fig. 2

Immunohistochemical staining detect the LT proteins in UTUC and adjacent tissues. Sections of UTUC tissues were immunostained using anti-LT monoclonal antibodies (a–f). Positive and negative control staining was performed using human prostate adenocarcinoma tissue sections. Positive control staining is shown in panel (a), while negative control staining is shown in panel (b). In UTUC tissue Sect. 33, positive staining for LT protein is observed (c), whereas in tissue Sect. 54, negative staining is evident (d). Similarly, in adjacent tissue Sect. 14, positive staining for LT protein is observed (e), while negative staining is observed in tissue Sect. 31 (f). Magnification, × 200. LT, large T; UTUC, upper tract urothelial carcinoma

Fig. 3

Immunohistochemical staining detect the p53 proteins in UTUC tissues. Sections of UTUC tissues were immunostained using anti-p53 monoclonal antibodies (a, a＇, b, b＇, c, c＇). In UTUC tissue Sect. 18, a scattered pattern of sporadic, scattered p53 nuclear staining within urothelial cells is observed, suggesting a normal/wild-type pattern (panel a, magnification × 40, a＇× 200). Two abnormal (mutant) patterns are observed; in tissue Sect. 48, the staining shows diffuse overexpression, with strong nuclear staining extending from the basal layer into the suprabasal layers of urothelial cells (panel b, magnification × 40, panel b＇× 200), tissue Sect. 67, null pattern (mutant) is present, with a complete absence of p53 staining in the tumor, while positivity is observed in the background inflammatory cells (panel c, magnification × 40, panel c＇× 200)