NON-INVASIVE BIOMARKERS IN BREAST CANCER EARLY DIAGNOSIS

Introduction: Breast cancer is the most commonly diagnosed malignancy among women worldwide and remains a leading cause of cancer-related death. Early detection plays a critical role in reducing mortality rates, improving prognosis and enhancing the effectiveness of therapeutic interventions. However, current screening strategies such as mammography and core needle biopsy (CNB) suffer from significant limitations, including reduced sensitivity in dense breast tissue, high false-positive rates and limited ability to characterize tumor biology at early stages. These limitations underscore the need for innovative, minimally invasive diagnostic tools. Liquid biopsy, which enables the analysis of tumor-derived biomarkers in body fluids, offers a promising alternative for early and accurate detection of malignancies. The integration of proteomic and epigenetic biomarkers derived from liquid biopsies could revolutionize early breast cancer diagnostics by improving precision and reducing reliance on invasive procedures. Methods: This work was conducted within the RENOVATE study, a prospective academic trial carried out at IRCCS Ospedale Policlinico San Martino (Genoa, Italy). Women presenting with suspicious breast lesions (BI-RADS 3–5, ≤2 cm) and healthy controls with negative mammograms were enrolled. Blood and urine samples were collected from all participants. Plasma and cfDNA were analyzed using a multi-omics approach combining high-throughput proteomic platforms (SomaScan™, Olink®), ELISA validation assays and epigenomic profiling techniques including cfMeDIP-seq and Oxford Nanopore sequencing. The primary objective was to identify circulating biomarkers capable of distinguishing breast cancer from benign conditions. Results: Multi-omics analysis revealed distinct molecular profiles in cancer patients compared to those with benign lesions or healthy controls. Proteomic profiling identified significantly altered levels of specific circulating proteins, including HPGDS, which demonstrated strong discriminative power. Epigenetic analysis of cfDNA via cfMeDIP-seq uncovered differentially methylated regions (DMRs) associated with genes involved in tumorigenesis, immune regulation, and cellular proliferation. These findings support the potential of circulating biomarkers to serve as a non-invasive diagnostic signature for early-stage breast cancer. Conclusions: This study provides compelling evidence for the clinical utility of liquid biopsy in early breast cancer diagnosis. The combination of proteomic and epigenetic signatures derived from blood samples enables molecular-level discrimination between malignant and benign breast lesions, offering a non-invasive, accurate, and patient-friendly diagnostic tool. By reducing unnecessary biopsies and enabling earlier therapeutic intervention, this approach has the potential to improve patient outcomes and support the broader implementation of precision oncology in clinical practice. Future larger-scale validation studies are essential to confirm these findings and transition liquid biopsy-based diagnostics into routine breast cancer screening protocols.