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

Martijn van der Heijden; Paul B.M. Essers; Monique C. de Jong; Reinout H. de Roest; Sebastian Sanduleanu; Caroline V.M. Verhagen; Olga Hamming-Vrieze; Frank Hoebers; Philippe Lambin; Harry Bartelink; C. René Leemans; Marcel Verheij; Ruud H. Brakenhoff; Michiel W.M. van den Brekel; Conchita Vens

doi:10.3389/fonc.2019.01470

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

Martijn van der Heijden, Paul B.M. Essers, Monique C. de Jong, Reinout H. de Roest, Sebastian Sanduleanu, Caroline V.M. Verhagen, Olga Hamming-Vrieze, Frank Hoebers, Philippe Lambin, Harry Bartelink, C. René Leemans, Marcel Verheij, Ruud H. Brakenhoff, Michiel W.M. van den Brekel, Conchita Vens

Maxillofacial Surgery (AMC)

Research output: Contribution to Journal › Article › Academic › peer-review

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). 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⁺ T-cell 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.

Original language	English
Article number	1470
Journal	Frontiers in oncology
Volume	9
DOIs	https://doi.org/10.3389/fonc.2019.01470
Publication status	Published - 10 Jan 2020

Bibliographical note

Funding Information:
The authors are grateful for financial support from Brunel and Verwelius and would like to thank the NKI Genomics Core Facility for performing RNA sequencing, the NKI RHPC facility for providing computational resources and the Core Facility-Molecular Pathology and Biobank (CFMPB) for collecting and preparing tissue samples. Funding. Authors acknowledge financial support from the Dutch Cancer Society (KWF-A6C7072, acronym DESIGN), from the EU 7th framework program (no 257144, acronym ARTFORCE) and the ERC advanced grant (no 694812, acronym Hypoximmuno).

Publisher Copyright:
© Copyright © 2020 van der Heijden, Essers, de Jong, de Roest, Sanduleanu, Verhagen, Hamming-Vrieze, Hoebers, Lambin, Bartelink, Leemans, Verheij, Brakenhoff, van den Brekel and Vens.

Funding

The authors are grateful for financial support from Brunel and Verwelius and would like to thank the NKI Genomics Core Facility for performing RNA sequencing, the NKI RHPC facility for providing computational resources and the Core Facility-Molecular Pathology and Biobank (CFMPB) for collecting and preparing tissue samples. Funding. Authors acknowledge financial support from the Dutch Cancer Society (KWF-A6C7072, acronym DESIGN), from the EU 7th framework program (no 257144, acronym ARTFORCE) and the ERC advanced grant (no 694812, acronym Hypoximmuno).

Funders	Funder number
Dutch Cancer Society	KWF-A6C7072
EU 7th framework program
Horizon 2020 Framework Programme	694812, 257144
European Research Council

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.3389/fonc.2019.01470

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van der Heijden, M., Essers, P. B. M., de Jong, M. C., de Roest, R. H., Sanduleanu, S., Verhagen, C. V. M., Hamming-Vrieze, O., Hoebers, F., Lambin, P., Bartelink, H., Leemans, C. R., Verheij, M., Brakenhoff, R. H., van den Brekel, M. W. M., & Vens, C. (2020). Biological Determinants of Chemo-Radiotherapy Response in HPV-Negative Head and Neck Cancer: A Multicentric External Validation. Frontiers in oncology, 9, Article 1470. https://doi.org/10.3389/fonc.2019.01470

@article{d578b9bf91bf4ca288e123011aa4bf41,

title = "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). 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+ T-cell 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.",

author = "{van der Heijden}, Martijn and Essers, {Paul B.M.} and {de Jong}, {Monique C.} and {de Roest}, {Reinout H.} and Sebastian Sanduleanu and Verhagen, {Caroline V.M.} and Olga Hamming-Vrieze and Frank Hoebers and Philippe Lambin and Harry Bartelink and Leemans, {C. Ren{\'e}} and Marcel Verheij and Brakenhoff, {Ruud H.} and {van den Brekel}, {Michiel W.M.} and Conchita Vens",

note = "Funding Information: The authors are grateful for financial support from Brunel and Verwelius and would like to thank the NKI Genomics Core Facility for performing RNA sequencing, the NKI RHPC facility for providing computational resources and the Core Facility-Molecular Pathology and Biobank (CFMPB) for collecting and preparing tissue samples. Funding. Authors acknowledge financial support from the Dutch Cancer Society (KWF-A6C7072, acronym DESIGN), from the EU 7th framework program (no 257144, acronym ARTFORCE) and the ERC advanced grant (no 694812, acronym Hypoximmuno). Publisher Copyright: {\textcopyright} Copyright {\textcopyright} 2020 van der Heijden, Essers, de Jong, de Roest, Sanduleanu, Verhagen, Hamming-Vrieze, Hoebers, Lambin, Bartelink, Leemans, Verheij, Brakenhoff, van den Brekel and Vens.",

year = "2020",

month = jan,

day = "10",

doi = "10.3389/fonc.2019.01470",

language = "English",

volume = "9",

journal = "Frontiers in oncology",

issn = "2234-943X",

publisher = "Frontiers Media S.A.",

}

van der Heijden, M, Essers, PBM, de Jong, MC, de Roest, RH, Sanduleanu, S, Verhagen, CVM, Hamming-Vrieze, O, Hoebers, F, Lambin, P, Bartelink, H, Leemans, CR, Verheij, M, Brakenhoff, RH, van den Brekel, MWM & Vens, C 2020, 'Biological Determinants of Chemo-Radiotherapy Response in HPV-Negative Head and Neck Cancer: A Multicentric External Validation', Frontiers in oncology, vol. 9, 1470. https://doi.org/10.3389/fonc.2019.01470

TY - JOUR

T1 - Biological Determinants of Chemo-Radiotherapy Response in HPV-Negative Head and Neck Cancer

T2 - A Multicentric External Validation

AU - van der Heijden, Martijn

AU - Essers, Paul B.M.

AU - de Jong, Monique C.

AU - de Roest, Reinout H.

AU - Sanduleanu, Sebastian

AU - Verhagen, Caroline V.M.

AU - Hamming-Vrieze, Olga

AU - Hoebers, Frank

AU - Lambin, Philippe

AU - Bartelink, Harry

AU - Leemans, C. René

AU - Verheij, Marcel

AU - Brakenhoff, Ruud H.

AU - van den Brekel, Michiel W.M.

AU - Vens, Conchita

N1 - Funding Information: The authors are grateful for financial support from Brunel and Verwelius and would like to thank the NKI Genomics Core Facility for performing RNA sequencing, the NKI RHPC facility for providing computational resources and the Core Facility-Molecular Pathology and Biobank (CFMPB) for collecting and preparing tissue samples. Funding. Authors acknowledge financial support from the Dutch Cancer Society (KWF-A6C7072, acronym DESIGN), from the EU 7th framework program (no 257144, acronym ARTFORCE) and the ERC advanced grant (no 694812, acronym Hypoximmuno). Publisher Copyright: © Copyright © 2020 van der Heijden, Essers, de Jong, de Roest, Sanduleanu, Verhagen, Hamming-Vrieze, Hoebers, Lambin, Bartelink, Leemans, Verheij, Brakenhoff, van den Brekel and Vens.

PY - 2020/1/10

Y1 - 2020/1/10

N2 - 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). 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+ T-cell 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.

AB - 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). 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+ T-cell 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.

U2 - 10.3389/fonc.2019.01470

DO - 10.3389/fonc.2019.01470

M3 - Article

SN - 2234-943X

VL - 9

JO - Frontiers in oncology

JF - Frontiers in oncology

M1 - 1470

ER -

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

Abstract

Bibliographical note

Funding

UN SDGs

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