Validation of a 15-gene hypoxia classifier in head and neck cancer for prospective use in clinical trials

• Published in: Acta oncologica Vol. 55; no. 9-10; p. 1091
• Main Authors: Toustrup, Kasper, Sørensen, Brita Singers, Metwally, Mohamed A Hassan, Tramm, Trine, Mortensen, Lise Saksø, Overgaard, Jens, Alsner, Jan
• Format: Journal Article
• Language: English
• Published: England 02-10-2016
• Subjects: Adult; Aged; Aged, 80 and over; Animals; Biomarkers, Tumor - genetics; Biopsy; Carcinoma, Squamous Cell - genetics; Carcinoma, Squamous Cell - pathology; Carcinoma, Squamous Cell - therapy; Cell Hypoxia - drug effects; Cell Hypoxia - genetics; Chemoradiotherapy - methods; Clinical Trials as Topic; Female; Gene Expression Profiling; Head and Neck Neoplasms - genetics; Head and Neck Neoplasms - pathology; Head and Neck Neoplasms - therapy; Humans; Male; Mice; Mice, Nude; Middle Aged; Nimorazole - therapeutic use; Radiation Tolerance - drug effects; Radiation Tolerance - physiology; Radiation-Sensitizing Agents - therapeutic use; Reproducibility of Results; Squamous Cell Carcinoma of Head and Neck; Xenograft Model Antitumor Assays
• ISSN: 1651-226X
• DOI: 10.3109/0284186X.2016.1167959

In head and neck squamous cell carcinomas (HNSCC) hypoxic radioresistance can be reduced by use of the hypoxic modifier nimorazole, as shown in the DAHANCA 5 trial. Recently, a 15-gene hypoxia classifier has shown predictive impact for the effect of nimorazole by identifying 'more' and 'less' hypoxic tumors in the DAHANCA 5 cohort. A prospective multicentre EORTC-1219 study is initiated, where nimorazole and prospective use of the classifier as a predictor is tested in relation to the most recent accelerated chemoradiotherapy treatment. Validation of the gene expression classification procedures is described here.
Formalin-fixed paraffin-embedded (FFPE) tumor material from three recent HNSCC cohorts [DAHANCA 18 (n = 96), 24 (n = 40), and IAEA Hypo (n = 55)] was used to establish and validate procedures for prospective classification of patients. Repeatability was tested for the different steps in the gene expression analysis, and reproducibility was tested with xenograft tumors (FaDu
, UTSCC33), where gene expression in complementary sections was compared after fixation and embedding locally and at international institutions, respectively. Intra-tumor heterogeneity was addressed by classifying biopsy samples from HNSCC tumors, where 2-4 biopsies from each tumor was accessible.
Procedures were successfully established for individual classification of HNSCC patients in retrospective and prospective cohorts. Measurements of gene expression levels were reproducible between different international institutions.
Technical validation of the 15-gene hypoxia classifier demonstrated that it is suitable for implementation in prospective clinical trials.