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. 2023 Nov;149(14):12597-12604.
doi: 10.1007/s00432-023-05077-3. Epub 2023 Jul 14.

Digital droplet PCR-based quantification of ccfHPV-DNA as liquid biopsy in HPV-driven cervical and vulvar cancer

Fabinshy Thangarajah  1   2 , Jana Busshoff  3 , Janina Salamon  3 , Marie-Sandrine Pruss  3 , Caroline Lenz  3 , Bernd Morgenstern  3 , Martin Hellmich  4 , Hans Anton Schlößer  5 , Maximilian Lenz  6 , Christian Domröse  3 , Michael R Mallmann  3 , Peter Mallmann  3 , Jonathan Weiß  7 , Fabian Franzen  7 , Sabine Merkelbach-Bruse  8 , Elke Binot  8 , Marie-Lisa Eich  8 , Reinhardt Büttner  8 , Anne Maria Schultheis #  9 , Christina Alidousty #  8
Affiliations

Digital droplet PCR-based quantification of ccfHPV-DNA as liquid biopsy in HPV-driven cervical and vulvar cancer

Fabinshy Thangarajah et al. J Cancer Res Clin Oncol. 2023 Nov.

Abstract

Purpose: More than 99% of cervical cancers and up to 40% of vulvar cancers are human papillomavirus (HPV) related. HPV 16 and 18 are the most relevant subtypes. Novel technologies allow the detection of minimal amounts of circulating cell-free HPV DNA (ccfHPV-DNA). The aim of this study was to evaluate ccfHPV-DNA assessed by droplet digital PCR (ddPCR) as a biomarker for molecular therapy monitoring in early, advanced, relapsed and metastatic HPV-driven cervical and vulvar cancer.

Methods: Inclusion criteria of the study were histologically proven HPV 16/18-driven cervical and vulvar cancer with first diagnosed disease, newly diagnosed recurrence, or progression of disease. Blood samples were taken pre- and post-therapeutically. Circulating cell-free HPV DNA was quantified using ddPCR and the results were correlated with clinical data.

Results: The mean copy number of ccfHPV-DNA was 838.6 (± 3089.1) in pretreatment and 2.3 (± 6.4) in post-treatment samples (p < 0.05). The copy number of ccfHPV-DNA increased with higher FIGO stages (p < 0.05), which are commonly used for clinical staging/assessment. Furthermore, we compared the distribution of copy numbers between T-stage 1 versus T-stage 2/3. We could show higher copy number level of ccfHPV-DNA in T-stage 2/3 (p < 0.05).

Conclusions: Therapy monitoring with determination of ccfHPV-DNA by ddPCR with a small amount of plasma reflects response to therapy and appears feasible for patients in advanced cancer stages of cervical and vulvar cancer. This promising tool should be examined as marker of therapy monitoring in particular in novel HPV-directed therapies.

Keywords: Cervical cancer; Liquid biopsy; Vulvar cancer; cHPV; cfDNA; ddPCR.

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Conflict of interest statement

The authors have no conflict of interest to declare.

Figures

Fig. 1
Fig. 1
Illustration of pre- and post-therapeutic ccfHPV-DNA level (copies per ml plasma). Wilcoxon matched-pairs signed rank test for paired comparisons of pre- and post-therapeutic samples. Significant difference is indicated by asterisks. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001
Fig. 2
Fig. 2
Comparison of mean copy number ccfHPV-DNA level per ml plasma between Figo stages I/II versus Figo stages III/IV. Significant differences were calculated using the nonparametric, unpaired and two-tailed Mann–Whitney test for unpaired comparisons. Significant difference is indicated by asterisks. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001
Fig. 3
Fig. 3
Comparison of mean copy number ccfHPV-DNA per ml plasma between T1 versus T2/3. Significant differences were calculated using the nonparametric, unpaired and two-tailed Mann–Whitney test for unpaired comparisons. Significant difference is indicated by asterisks. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001
Fig. 4
Fig. 4
Dynamics of ccfHPV-DNA during therapy of first diagnosed cervical cancer. This figure shows patients with FIGO IIIC stage (A) und FIGO IVA stage (B) cervical cancer. It shows the copy number at baseline (1), after lymphadenectomy (2) and induction chemotherapy followed by radiochemotherapy (3). In both patients, ccfHPV-DNA was not detectable after the therapy
See this image and copyright information in PMC

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