An Integrated Approach Identifies Mediators of Local Recurrence in Head and Neck Squamous Carcinoma

Clin Cancer Res. 2017 Jul 15;23(14):3769-3780. doi: 10.1158/1078-0432.CCR-16-2814. Epub 2017 Feb 7.


Purpose: Head and neck squamous cell carcinomas (HNSCCs) cause more than 300,000 deaths worldwide each year. Locoregional and distant recurrences represent worse prognostic events and accepted surrogate markers of patients' overall survival. No valid biomarker and salvage therapy exist to identify and treat patients at high-risk of recurrence. We aimed to verify if selected miRNAs could be used as biomarkers of recurrence in HNSCC.Experimental Design: A NanoString array was used to identify miRNAs associated with locoregional recurrence in 44 patients with HNSCC. Bioinformatic approaches validated the signature and identified potential miRNA targets. Validation experiments were performed using an independent cohort of primary HNSCC samples and a panel of HNSCC cell lines. In vivo experiments validated the in vitro results.Results: Our data identified a four-miRNA signature that classified HNSCC patients at high- or low-risk of recurrence. These miRNAs collectively impinge on the epithelial-mesenchymal transition process. In silico and wet lab approaches showed that miR-9, expressed at high levels in recurrent HNSCC, targets SASH1 and KRT13, whereas miR-1, miR-133, and miR-150, expressed at low levels in recurrent HNSCC, collectively target SP1 and TGFβ pathways. A six-gene signature comprising these targets identified patients at high risk of recurrences, as well. Combined pharmacological inhibition of SP1 and TGFβ pathways induced HNSCC cell death and, when timely administered, prevented recurrence formation in a preclinical model of HNSCC recurrence.Conclusions: By integrating different experimental approaches and competences, we identified critical mediators of recurrence formation in HNSCC that may merit to be considered for future clinical development. Clin Cancer Res; 23(14); 3769-80. ©2017 AACR.

MeSH terms

  • Adult
  • Aged
  • Animals
  • Biomarkers, Tumor / genetics
  • Carcinoma, Squamous Cell / genetics*
  • Carcinoma, Squamous Cell / pathology
  • Cell Line, Tumor
  • Epithelial-Mesenchymal Transition / genetics
  • Female
  • Gene Expression Regulation, Neoplastic
  • Head and Neck Neoplasms / genetics*
  • Head and Neck Neoplasms / pathology
  • Humans
  • Keratin-13 / genetics
  • Male
  • Mice
  • MicroRNAs / genetics*
  • Middle Aged
  • Neoplasm Recurrence, Local / genetics*
  • Neoplasm Recurrence, Local / pathology
  • Signal Transduction
  • Sp1 Transcription Factor / genetics
  • Squamous Cell Carcinoma of Head and Neck
  • Transforming Growth Factor beta / genetics
  • Tumor Suppressor Proteins / genetics
  • Xenograft Model Antitumor Assays


  • Biomarkers, Tumor
  • KRT13 protein, human
  • Keratin-13
  • MIRN1 microRNA, human
  • MIRN133 microRNA, human
  • MIRN150 microRNA, human
  • MIRN92 microRNA, human
  • MicroRNAs
  • SASH1 protein, human
  • Sp1 Transcription Factor
  • SP1 protein, human
  • Transforming Growth Factor beta
  • Tumor Suppressor Proteins