Identification of potential prognostic and predictive biomarkers in Head and Neck Carcinoma

Head and neck cancer (HNC) represents the seventh most common malignancy worldwide and is a heterogeneous disease affecting the upper aerodigestive tract. The majority of cases (>90%) are squamous cell carcinomas (SCC), whereas a minority originate from salivary glands, lymphatic tissue, or other less common components. Among the subtypes, laryngeal carcinoma (LCa) is the most prevalent, accounting for approximately one third of all HNC cases, with around 189,000 new diagnoses each year. Advanced stages are frequently associated with cervical lymph node metastases, the primary sites of dissemination, whose involvement represents a crucial prognostic factor and significantly impacts patient survival. Recent studies have highlighted that the combined analysis of gene expression and deregulated miRNAs is a powerful tool to better understand tumor progression and identify novel therapeutic targets, thereby improving diagnostic strategies and personalized treatments. MicroRNAs (miRNAs) play a fundamental role in the post-transcriptional regulation of gene expression and the control of cell fate. In this study, a cohort of patients with laryngeal carcinoma was analyzed through transcriptomic profiling, leading to the identification of deregulated genes and miRNAs potentially associated with disease progression. NGS analyses revealed the Relaxin Family Peptide Receptor 1 (RXFP1) as a potential biomarker associated with advanced stages of the disease, suggesting that its upregulation in patients with lymph node metastases (N+) may reflect and predict tumor progression. To further investigate the functional role of RXFP1, two stable models were generated in HNO-210 cell lines through third-generation lentiviral transduction: one carrying an empty backbone vector and one with constitutive receptor overexpression. The main findings of this study highlight an increase in cAMP levels following RXFP1 activation mediated by RLN2. This increase is associated with modulation of the MAPK pathway and a concomitant reduction in the phosphorylation of downstream effectors of the PI3K/AKT pathway. These results suggest that cAMP activation mediated by RXFP1 may compensate for the decreased AKT activity through ERK activation, promoting a pro-migratory effect. This phenomenon favors a partial cellular plasticity phenotype, characterized by increased migration and invasion post-stimulation, associated with the modulation of matrix metalloproteinase 9 as the main downstream effector.