Targeted next-generation sequencing identifies molecular subgroups in squamous cell carcinoma of the head and neck with distinct outcome after concurrent chemoradiation.
Based on epidemiological (HPV status, smoking habits) and clinical risk factors (T/N stage), three subgroups of patients suffering from locally advanced oropharyngeal carcinoma with significantly different outcome after concurrent chemoradiation (cCRTX) can be distinguished. Mutational profiling by targeted next-generation sequencing (NGS) might further improve risk stratification.



Patients with stage IV squamous cell carcinoma of the oropharynx and hypopharynx who had been enrolled in a randomized phase III trial (ARO-0401) comparing two regimens of cCRTX and from whom archival tumor specimens were available were included. The HPV status was determined by p16 immunostaining and detection of HPV DNA. Targeted NGS covering 45 genes frequently altered in squamous cell carcinoma of the head and neck (SCCHN) was applied for detection of non-synonymous somatic and germline mutations. Interference of mutational profiles with cCRTX efficacy was determined.



The prognostic value of the 'Ang' risk model could be confirmed in the total biomarker study cohort (N = 175) as well as the patient subgroup for which mutational profiles could be established (N = 97). Mutations in genes involved in phosphoinositide 3-kinase (PI3K), receptor tyrosine kinase (RTK), and p53 signaling pathways were significantly enriched in the low- (N = 7), intermediate- (N = 20), and high-risk group (N = 70), respectively. Mutations in TP53 identified a subgroup of high-risk patients with dismal outcome after cCRTX. No prognostic relevance was observed for mutations in PI3K and RTK signaling pathways in the low- and intermediate-risk groups, respectively. Mutated NOTCH1 and two functional KDR germline variants (rs2305948, rs1870377) were associated with improved outcome in all risk groups. All genetic markers (TP53, NOTCH1, KDR) remained independent prognosticators of OS in the multivariate model.



A potential of targeted NGS for risk classification of SCCHN cases beyond HPV status and clinical factors was demonstrated.
Department of Radiooncology and Radiotherapy, Charite University Hospital, Berlin ingeborg.tinhofer@charite.de.|German Cancer Research Center (DKFZ), Heidelberg, and German Cancer Consortium (DKTK) Partner Site Berlin.
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Tinhofer
Institute of Pathology, University Hospital and National Center for Tumor Diseases, Heidelberg, Germany.|Department of Pathology, Center for Integrated Diagnostics, Massachusetts General Hospital/Harvard Medical School, Boston, USA.
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A
Stenzinger
Department of Radiooncology and Radiotherapy, Charite University Hospital, Berlin.|German Cancer Research Center (DKFZ), Heidelberg, and German Cancer Consortium (DKTK) Partner Site Berlin.
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T
Eder
Department of Radiooncology and Radiotherapy, Charite University Hospital, Berlin.|German Cancer Research Center (DKFZ), Heidelberg, and German Cancer Consortium (DKTK) Partner Site Berlin.
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Konschak
Department of Radiooncology and Radiotherapy, Charite University Hospital, Berlin.|German Cancer Research Center (DKFZ), Heidelberg, and German Cancer Consortium (DKTK) Partner Site Berlin.
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F
Niehr
Institute of Pathology, University Hospital and National Center for Tumor Diseases, Heidelberg, Germany.
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Endris
Department of Radiation Oncology, University Hospitals Erlangen-Nürnberg, Erlangen.
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Distel
Department of Radiotherapy, University Hospital Regensburg.
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Hautmann
Department of Otorhinolaryngology, Head & Neck Surgery, University Hospital Giessen-Marburg, Marburg.
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Mandic
Department of Radiooncology and Radiotherapy, Charite University Hospital, Berlin.
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C
Stromberger
Institute of Pathology, University Hospital and National Center for Tumor Diseases, Heidelberg, Germany.|Institute of Pathology, Technical University Munich (TUM), Munich.|German Cancer Research Center (DKFZ), Heidelberg, and German Cancer Consortium (DKTK) partner site Munich, Germany.
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Weichert
Department of Radiooncology and Radiotherapy, Charite University Hospital, Berlin.
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V
Budach
D000368:Aged; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology / Q000532:radiotherapy*; D059248:Chemoradiotherapy; D018572:Disease-Free Survival; D005260:Female; D006258:Head and Neck Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology / Q000532:radiotherapy*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics*; D009364:Neoplasm Recurrence, Local / Q000188:drug therapy / Q000235:genetics / Q000473:pathology / Q000532:radiotherapy; D009367:Neoplasm Staging; D027383:Papillomaviridae / Q000235:genetics / Q000472:pathogenicity; D011379:Prognosis; D015398:Signal Transduction; D000077195:Squamous Cell Carcinoma of Head and Neck
D009363:Neoplasm Proteins
concurrent chemoradiation; hypopharyngeal carcinoma; molecular risk stratification; mutation profile; oropharyngeal carcinoma; targeted next-generation sequencing