Characterization of a head and neck cancer-derived cell line panel confirms the distinct TP53-proficient copy number-silent subclass

Oral Oncol. 2019 Nov:98:53-61. doi: 10.1016/j.oraloncology.2019.09.004. Epub 2019 Sep 18.

Abstract

Introduction: Head and neck squamous cell carcinomas (HNSCC) arise in the mucosal lining of the upper aerodigestive tract. Risk factors are exogenous carcinogen exposure, human papillomavirus (HPV) infection, and genetic predisposition such as Fanconi anemia (FA). Clinically, tumors are stratified based on stage, site and HPV-status. The majority of HPV-positive and -negative HNSCC is characterized by frequent copy number (CN) changes and an abrogated p53-pathway. A third genetically-defined HPV-negative subclass of HNSCC is emerging: tumors that lack gross chromosomal changes (CN-silent), are mostly TP53-proficient, and have a relatively favorable prognosis.

Methods: A representative panel of HPV-positive, HPV-negative and FA-HNSCC-derived cell lines was genetically characterized.

Results: Despite apparent differences in etiology, FA-HNSCC cell lines show comparable genetic alterations as sporadic non-FA-HNSCC-derived cell lines. Furthermore, we identified a near diploid CN-silent HPV-negative HNSCC line: VU-SCC-040. Molecular characterization uncovers the absence of TP53 mutations, a functional p53-pathway and a CASP8 mutation. TP53 gene knockout using CRISPR-Cas9 resulted in resistance to MDM2 inhibition. Whereas p53-status is often proposed as a predictive biomarker for treatment response, TP53-knockout did not change sensitivity to cisplatin, Chk1 and Wee1 inhibition. Additionally, 84 CN-silent tumors were identified in the HNSCC PanCancer cohort and shown to be enriched for female gender, HRAS and CASP8 mutations.

Conclusion: FA-derived HNSCC cell lines share comparable CN-profiles and mutation patterns as sporadic HPV-negative HNSCC. In contrast, a subclass of CN-silent, HPV-negative and TP53 wild-type HNSCC separates from the majority of HNSCC tumors. We show that VU-SCC-040 is a HNSCC cell model representative of this subclass.

Keywords: Copy number silent; Fanconi anemia; Head and neck squamous cell carcinoma; Low-coverage whole genome sequencing; TP53 wild-type; Target-enrichment sequencing; Targeted treatment; p53 pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers, Tumor
  • Cell Line, Tumor
  • DNA Copy Number Variations*
  • Gene Knockout Techniques
  • Head and Neck Neoplasms / etiology
  • Head and Neck Neoplasms / genetics*
  • Humans
  • Mutation*
  • Papillomavirus Infections / complications
  • Squamous Cell Carcinoma of Head and Neck / genetics
  • Tumor Suppressor Protein p53 / genetics*
  • Whole Genome Sequencing

Substances

  • Biomarkers, Tumor
  • TP53 protein, human
  • Tumor Suppressor Protein p53