Evaluation of Integrated HPV DNA as Individualized Biomarkers for the Detection of Recurrent CIN2/3 during Post-Treatment Surveillance

Abstract

Purpose: Post-treatment follow-up in women with cervical pre-cancers (CIN3) is mandatory due to relapse in up to 10% of patients. Standard follow-up based on hrHPV-DNA/cytology co-testing has high sensitivity but limited specificity. The aim of our prospective, multicenter, observational study was to test the hypothesis that an individualized viral-cellular-junction test (vcj-PCR) combined with cytology has a lower false positive rate for the prediction of recurrence compared to standard co-testing. Methods: Pre-surgical cervical swabs served for the identification of HPV16/18 DNA integration sites by next-generation-sequencing (NGS). Samples taken at 6, 12 and 24 months post-surgery were evaluated by cytology, hrHPV-DNA and the patients’ individual HPV-integration sites (vcj-PCR on the basis of NGS). Results: Integration sites were detected in 48 of 445 patients (10.8%), 39 of them had valid follow-up data. The false positive rate was 18.2% (95% CI 8.6–34.4%) for standard hrHPV/cytology at six months compared to 12.1% (95% CI 4.8–27.3%) for vcj-PCR/cytology, respectively (McNemar p = 0.50). Six patients developed recurrences (1 CIN2, 5 CIN3) during follow-up. Standard co-testing detected all, whereas vcj-PCR/cytology detected only five patients with recurrences. Data of 269 patients without evidence of HPV16/18 integration were subject to post-hoc analyses. Standard co-testing revealed a false positive rate of 15.7% (95% CI 11.7–20.7%) and predicted ten of fourteen recurrences at six months. Conclusions: Although highly specific on its own vcj-PCR could not detect all recurrent CIN2/3. Possible reasons for this unexpected result may be multifocal lesions, intratumoral heterogeneity with respect to HPV integration and/or incident CIN.

Human papillomavirus associated cervical lesion: pathogenesis and therapeutic interventions

“…HPV integration into the host genome results from heightened genome instability in HPV‐infected cells 199 . The frequency of HPV integrations progressively rises during the progression of cervical lesions, 200–202 rendering it a plausible biomarker for the surveillance of cervical cancer and precancerous disease conditions 203–205 …”

“…199 The frequency of HPV integrations progressively rises during the progression of cervical lesions, [200][201][202] rendering it a plausible biomarker for the surveillance of cervical cancer and precancerous disease conditions. [203][204][205] HPV integration events transpire across all human chromosomes; however, a Chinese study unveiled an excessive concentration of these events on chromosome 19. 206 Although HPV integration might seem stochastic, recent studies have pinpointed a growing array of HPV integration hotspot genes, including FHIT, LRP1B, PP1R37, HECW2, EMBP1, ANKRD50, SPTBN4, LINC00895, LYRM4-AS1, LINC00374, RBFOX1, CSMD1, CDH13, KLHL4, KLF12, KLF5, CCDC106.…”

“…199 The frequency of HPV integrations progressively rises during the progression of cervical lesions, 200 , 201 , 202 rendering it a plausible biomarker for the surveillance of cervical cancer and precancerous disease conditions. 203 , 204 , 205 …”

Standardized Procedures for Patients with Dysplasia and Other Diseases of the Cervix, Vulva, and Vagina at a Certified Dysplasia Unit Prior to the Introduction of the Organized Cervical Cancer Screening Program

“…1 dargestellt [4,5] Wissenschaftliche Fragestellungen der Dysplasie-Einheit wurden über Publikationen zu individuellen Prognosefaktoren, wie Tumor Budding für die Entstehung von plaettenepithelialen Zervixkarzinomen, Integraten und Subtypisierungen von HPV, sowie die Übereinstimmung von externer und intern durchgeführter Zytologie, von kolposkopischer bzw. vulvoskopischer Eindrücke und der durchgeführten Histologie und die Genauigkeit von kolposkopisch durchgeführten Biopsien der Zervix, Vulva und Vagina veröffentlicht [6,7,8,9,10,11,12]. Diese wurden auf verschiedenen wissenschaftlichen Kongressen vorgestellt.…”

“…Scientific investigations of the dysplasia unit have been published on individual prognosis factors such as tumor budding in the development of squamous epithelial cervical carcinoma, HPV integration and subtyping, the consistency of external vs. in-house cytologic results, of colposcopic or vulvoscopic impressions and histology performed, and the accuracy of biopsies of the cervix, vulva and vagina performed by colposcopy 6 7 8 9 10 11 12 . This research has been presented at various scientific conferences.…”