A Five-MicroRNA Signature Predicts Survival and Disease Control of Patients with Head and Neck Cancer Negative for HPV-infection

Purpose: HPV-negative head and neck squamous cell carcinoma (HNSCC) associates with unfavorable prognosis while independent prognostic markers remain to be defined. Experimental Design: We retrospectively performed miRNA expression profiling. Patients were operated for locally advanced HPV-negative HNSCC and had received radiochemotherapy in eight different hospitals (DKTK-ROG; n=85). Selection fulfilled comparable demographic, treatment and follow-up characteristics. Findings were validated in an independent single-center patient sample (LMU-KKG; n=77). A prognostic miRNA-signature was developed for freedom from recurrence and tested for other endpoints. Recursive-partitioning analysis was performed on the miRNA-signature, tumor and nodal stage, and extracapsular nodal spread. Technical validation used qRT-PCR. A miRNA-mRNA target network was generated and analyzed. Results: For DKTK-ROG and LMU-KKG patients, the median follow-up was 5.1 and 5.3 years, the 5-year freedom from recurrence rate was 63.5% and 75.3%, respectively. A five-miRNA-signature (hsa-let-7g-3p, hsa-miR-6508-5p, hsa-miR-210-5p, hsa-miR-4306 and hsa-miR-7161-3p) predicted freedom from recurrence in DKTK-ROG (HR 4.42, 95% CI 1.98−9.88, P<0.001), which was confirmed in LMU-KKG (HR 4.24, 95% CI 1.40−12.81, P=0.005). The signature also predicted overall survival (HR 3.03, 95% CI 1.50−6.12, P=0.001), recurrence-free survival (HR 3.16, 95% CI 1.65−6.04, P<0.001) and disease-specific survival (HR 5.12, 95% CI 1.88−13.92, P<0.001), all confirmed in LMU-KKG data. Adjustment for relevant covariates maintained the miRNA-signature predicting all endpoints. Recursive-partitioning analysis of both samples combined classified patients into low (n=17), low-intermediate (n=80), high-intermediate (n=48) or high risk (n=17) for recurrence (P<0.001). Conclusions: The five-miRNA-signature is a strong and independent prognostic factor for disease recurrence and survival of patients with HPV-negative HNSCC.