The Diagnostic Value of Circulating Cell-Free HPV DNA in Plasma from Cervical Cancer Patients

Abstract

Circulating cell-free HPV DNA (ccfHPV DNA) may serve as a marker for cervical cancer. In this study, we used digital droplet PCR (ddPCR) to detect and quantify ccfHPV DNA in plasma from patients with HPV16- or HPV18-associated cervical cancer. Blood samples from 60 patients diagnosed with cervical cancer (FIGO IA1-IVA) at Aarhus or Odense University Hospital (June 2018 to March 2020) were collected prior to treatment, and patients were subdivided into an early stage (n = 30) and a late-stage subgroup (n = 30) according to disease stage. Furthermore, blood samples from eight women with HPV16- or18-associated premalignant conditions (CIN3), and 15 healthy controls were collected. ddPCR was used to analyze plasma from all participants. ccfHPV DNA was detected in 19 late-stage patients (63.33%), 3 early stage patients (10.00%), and none of the CIN3 patients or controls. Quantitative evaluation showed significant correlations between ccfHPV DNA level and stage, tumor score, and tumor size. Thus, our results indicate that ccfHPV DNA may not be a useful marker for early detection of cervical cancer. However, for patients with advanced stage cervical cancer, ccfHPV DNA level represents a promising tool to establish tumor burden, making it useful for establishing treatment response and monitoring the disease.

The current status of cell‐free human papillomavirus DNA as a biomarker in cervical cancer and other HPV‐associated tumors: A review

“…[75][76][77] ctHPV DNA is more likely to have a larger role when CC is already diagnosed. While a few studies have shown that pretreatment ctHPV DNA levels do not correlate to prognosis, 54,56,57 several studies have demonstrated that persistent end-of-treatment ctHPV DNA seems to be a negative prognostic factor in cervical cancer 54,57 as well as in other HPV-related malignancies. 70,71 The most important clinical utility of ctHPV DNA in the future might however be as a surveillance marker for early detection of relapse and a few studies indicate that ctHPV DNA might be superior to clinical examination and radiology.…”

“…55 Recently another two studies, stratifying between early-and advanced cervical cancer, found ctHPV DNA in the majority of advanced cases (94% and 63%, respectively), but in a minority of early-stage CC (26.7% and 10%, respectively). 56,57 T A B L E 1 Studies investigating detection of ctHPV DNA in pre-treatment liquid biopsies from patients with invasive cervical cancer 50,60 Of note, there was, surprisingly, rather poor correlation between HPV-status in plasma and in the cervix. 60 Thus, there are contradicting reports whether ctHPV DNA can be found in plasma from patients with precursor lesions.…”

“…On the other hand, the few recent studies using ddPCR have not found ctHPV DNA in any precursor lesion. 54,56,57 1.5 | ctHPV DNA detection in women without cervical pathology…”

“…Several studies report that pre‐treatment amounts of ctHPV DNA correlate to those of HPV DNA in the cervical tumors 52,54 as well as to more advanced tumor characteristics such as higher stage, 44,46,47,51,52,54,57 increased tumor burden 59 and larger tumor size, 43,51,61 although Yang et al did not find a correlation to FIGO‐stage 50 . As mentioned above, two recent studies stratifying CC into early‐ and advanced stages found ctHPV DNA in a majority of the advanced staged CC but only in a minority of early‐stage CC 56,57 . Furthermore, an association with pre‐treatment levels of ctHPV DNA and increased risk for lymph node metastasis 43,52,54,61 as well as distant metastasis 46,61 has been described.…”

“…Furthermore, Kang et al, detected ctHPV DNA in serum of 19/19 (100%) patients with HPV‐positive metastatic cervical cancer 55 . Recently another two studies, stratifying between early‐ and advanced cervical cancer, found ctHPV DNA in the majority of advanced cases (94% and 63%, respectively), but in a minority of early‐stage CC (26.7% and 10%, respectively) 56,57 …”