Liquid Biopsy in the Diagnosis of HPV DNA in Breast Lesions

Abstract

Aim: HPV DNA has never been investigated in nipple discharges (ND) and serum-derived extracellular vesicles, although its presence has been reported in ductal lavage fluids and blood specimens. Materials & methods: We analyzed 50 ND, 22 serum-derived extracellular vesicles as well as 51 pathologic breast tissues for the presence of 16 HPV DNA types. Results: We show that the presence of HPV DNA in the ND is predictive of HPV DNA-positive breast lesions and that HPV DNA is more represented in intraductal papillomas. We also show the presence of HPV DNA in the serum-derived extracellular vesicles. Conclusion: Our data supports the use of liquid biopsy to detect HPV DNA in breast pathology. The presence of HPV DNA in breast cancer has been widely investigated in the last three decades [1], but few data are available on its presence in benign breast lesions, in other words, intraductal papilloma (IP). A plethora of pathogens has been found in triple negative breast cancer, the most aggressive breast cancer phenotype, including mucosal and cutaneous HPVs [2]. The presence of HPV DNA has been reported in biological fluids such as milk and colostrum of lactating women as well as in ductal lavages from women at increased risk of breast cancer [3,4]. The presence of HPV DNA has never been investigated in nipple discharge, a spontaneous or induced biological fluid which is examined to exclude the presence of malignant cells in women with any palpable lump or mammographic abnormalities. Interestingly, HPV DNA has recently been described in the blood of cervical cancer affected women [5]. Recent literature also shows that viral nucleic acids are cargoed by circulating extracellular vesicles (EVs), which are membrane-derived vesicles involved in cell-to-cell communication [6,7].We first searched for the presence of 16 high-risk mucosal HPV DNA in the virome of breast tissues and their biological fluids, in other words, nipple discharges (ND) and serum-derived EVs, of women with benign or malignant breast cancer lesions. Then, we searched if a noninvasive approach, the liquid biopsy, could be useful in the diagnosis of HPV DNA-positive breast lesions.

HPV DNA Associates With Breast Cancer Malignancy and It Is Transferred to Breast Cancer Stromal Cells by Extracellular Vesicles

“…At this regard, some authors reported circulating HPV DNA in the serum of patients with HPV-associated invasive cancer (10) and as marker for disease extent and recurrence (14). We also found HPV DNA in the serum derived-extracellular vesicles (EVs) of breast pathologies affected women as well as in patient with HPV DNA positive squamous cell carcinoma of the middle rectum (15, 16). EVs are a heterogeneous population of vesicles, which includes exosomes, microvesicles, and apoptotic bodies that differ in size and biophysical properties (17).…”

Oral HPV16 Prevalence in Oral Potentially Malignant Disorders and Oral Cavity Cancers

“…To investigate the HPV type distribution in patients with OPMD and OC, a high-throughput MALDI-TOF mass spectrometry-based method (Mass Array Platform, Agena Bioscience, Hamburg, Germany) was used, as previously described [18]. This method is able to detect 16 HPV DNA types (HPV16, 18,31,33,35,39,45,51,52,53,56,58,59, 66, 68 and 73) in a single well.…”

Putative Role of Circulating Human Papillomavirus DNA in the Development of Primary Squamous Cell Carcinoma of the Middle Rectum: A Case Report

“…Previous studies on cervical, oropharingeal, and breast diseases demonstrated the presence of circulating HPV DNA in the blood of affected patients (8, 32–35). Accordingly, we searched HPV DNA also in the blood of our patient.…”

“…Tumor-derived exosomes and extracellular vesicles are garnering increasing attention because of their ability to transfer bioactive molecules (mRNAs, microRNAs, DNA, and proteins) between neighboring cancerous or normal cells, and to contribute to human cancer progression. Moreover, exosomes and extracellular vesicles have been implicated in HPV transmission and carcinogenesis (7), and the presence of HPV DNA in the serum derived extracellular vesicles (EVs) of patients with breast diseases has been recently reported (8).…”