Biological Determinants of Chemo-Radiotherapy Response in HPV-Negative Head and Neck Cancer: A Multicentric External Validation

Abstract

Purpose: Tumor markers that are related to hypoxia, proliferation, DNA damage repair and stem cell-ness, have a prognostic value in advanced stage HNSCC patients when assessed individually. Here we aimed to evaluate and validate this in a multifactorial context and assess interrelation and the combined role of these biological factors in determining chemo-radiotherapy response in HPV-negative advanced HNSCC. Methods: RNA sequencing data of pre-treatment biopsy material from 197 HPV-negative advanced stage HNSCC patients treated with definitive chemoradiotherapy was analyzed. Biological parameter scores were assigned to patient samples using previously generated and described gene expression signatures. Locoregional control rates were used to assess the role of these biological parameters in radiation response and compared to distant metastasis data. Biological factors were ranked according to their clinical impact using bootstrapping methods and multivariate Cox regression analyses that included clinical variables. Multivariate Cox regression analyses comprising all biological variables were used to define their relative role among all factors when combined. Results: Only few biomarker scores correlate with each other, underscoring their independence. The different biological factors do not correlate or cluster, except for the two stem cell markers CD44 and SLC3A2 (r = 0.4, p < 0.001) and acute hypoxia prediction scores which correlated with T-cell infiltration score, CD8 + T cell abundance and proliferation scores (r = 0.52, 0.56, and 0.6, respectively with p < 0.001). van der Heijden et al. HNSCC Biology Impacting Chemo-Radiotherapy Outcomes Locoregional control association analyses revealed that chronic (Hazard Ratio (HR) = 3.9) and acute hypoxia (HR = 1.9), followed by stem cell-ness (CD44/SLC3A2; HR = 2.2/2.3), were the strongest and most robust determinants of radiation response. Furthermore, multivariable analysis, considering other biological and clinical factors, reveal a significant role for EGFR expression (HR = 2.9, p < 0.05) and T-cell infiltration (CD8 + T-cells: HR = 2.2, p < 0.05; CD8 + T-cells/Treg: HR = 2.6, p < 0.01) signatures in locoregional control of chemoradiotherapy-treated HNSCC. Conclusion: Tumor acute and chronic hypoxia, stem cell-ness, and CD8 + Tcell parameters are relevant and largely independent biological factors that together contribute to locoregional control. The combined analyses illustrate the additive value of multifactorial analyses and support a role for EGFR expression analysis and immune cell markers in addition to previously validated biomarkers. This external validation underscores the relevance of biological factors in determining chemoradiotherapy outcome in HNSCC.

Epithelial-to-mesenchymal transition is a prognostic marker for patient outcome in advanced stage HNSCC patients treated with chemoradiotherapy

“…The role of TNM, in contrast to EMT, in outcome is small and insignificant in both cohorts (Sup-Table S3 and S7), likely due to the advanced stage [4]. Multivariate analyses that include T-stage and N-stage provide increased HR and confirm the EMT association with OS and PFS in both cohorts (p < 0.05; Fig.…”

“…Tumor biology studies revealed an important role for HPV and hypoxia in HNSCC [2][3][4]. HPV-positive oropharyngeal tumors represent a distinct subgroup of HNSCC that is associated with a good prognosis whereas tumor hypoxia markers reveal patients with a poor prognosis [5][6][7].…”

Mass Spectrometric Comparison of HPV-Positive and HPV-Negative Oropharyngeal Cancer

“…Beyond or as a consequence of amino acid metabolism, SLC3A2/CD98 are involved in a wide range of functions relevant for cancer development and survival, such as mTOR pathway activation and autophagy [ 58 ], integrin signaling [ 59 ], immunity [ 60 ] and oxidative stress [ 58 , 61 ] and are negative prognostic factors in various cancers [ 62 , 63 ]. In HNSCC, SLC3A2/CD98 were described to mark tumor stem cell populations [ 64 ] and to confer a negative prognosis, which was completely restricted to HPV-positive tumors in one report [ 65 ] but primarily described for HPV-negative tumors in others [ 58 , 66 , 67 , 68 ]. In contrast to SLC3A2, there are no data regarding a possible prognostic role of leucine-rich PPR motif-containing protein (LRPPRC) in HNSCC.…”

A Combined Prediction Model for Lymph Node Metastasis Based on a Molecular Panel and Clinicopathological Factors in Oral Squamous Cell Carcinoma

“…Many studies have found that OSCC is a polygenic disease, and gene expression profiling technology has made highthroughput gene analysis possible (10)(11)(12)(13). Researchers can obtain the gene expression characteristics of a certain type of tumor by analyzing the gene expression profiles of tumor samples (14). Our previous research detected the expression of 22 candidate genes and 1 housekeeping gene in 120 OSCC tissue samples and 120 normal tissue samples at the mRNA level using real-time PCR (15).…”