SMAD4 loss is associated with cetuximab resistance and induction of MAPK/JNK activation in head and neck cancer cells

Hiroyuki Ozawa, Ruchira S. Ranaweera, Evgeny Izumchenko, Eugene Makarev, Alex Zhavoronkov, Elana J. Fertig, Jason D. Howard, Ana Markovic, Atul Bedi, Rajani Ravi, Jimena Perez, Quynh Thu Le, Christina S. Kong, Richard C. Jordan, Hao Wang, Hyunseok Kang, Harry Quon, David Sidransky, Christine H. Chung

Research output: Contribution to journalArticle

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Abstract

Purpose: We previously demonstrated an association between decreased SMAD4 expression and cetuximab resistance in head and neck squamous cell carcinoma (HNSCC). The purpose of this study was to further elucidate the clinical relevance of SMAD4 loss in HNSCC. Experimental Design: SMAD4 expression was assessed by IHC in 130 newly diagnosed and 43 patients with recurrent HNSCC. Correlative statistical analysis with clinicopathologic data was also performed. OncoFinder, a bioinformatics tool, was used to analyze molecular signaling in TCGA tumors with low or high SMAD4 mRNA levels. The role of SMAD4 was investigated by shRNA knockdown and gene reconstitution of HPV-negative HNSCC cell lines in vitro and in vivo. Results: Our analysis revealed that SMAD4 loss was associated with an aggressive, HPV-negative, cetuximab-resistant phenotype. We found a signature of prosurvival and antiapoptotic pathways that were commonly dysregulated in SMAD4-low cases derived from TCGA-HNSCC dataset and an independent oral cavity squamous cell carcinoma (OSCC) cohort obtained from GEO. We show that SMAD4 depletion in an HNSCC cell line induces cetuximab resistance and results in worse survival in an orthotopic mouse model in vivo. We implicate JNK and MAPK activation as mediators of cetuximab resistance and provide the foundation for the concomitant EGFR and JNK/MAPK inhibition as a potential strategy for overcoming cetuximab resistance in HNSCCs with SMAD4 loss. Conclusions: Our study demonstrates that loss of SMAD4 expression is a signature characterizing the cetuximab-resistant phenotype and suggests that SMAD4 expression may be a determinant of sensitivity/resistance to EGFR/MAPK or EGFR/JNK inhibition in HPV-negative HNSCC tumors.

Original languageEnglish
Pages (from-to)5162-5175
Number of pages14
JournalClinical Cancer Research
Volume23
Issue number17
DOIs
Publication statusPublished - 2017 Sep 1
Externally publishedYes

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Head and Neck Neoplasms
Gene Knockdown Techniques
Phenotype
Cell Line
Carcinoma, squamous cell of head and neck
Cetuximab
Computational Biology
Small Interfering RNA
Mouth
Squamous Cell Carcinoma
Neoplasms
Research Design
Messenger RNA
Survival

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

SMAD4 loss is associated with cetuximab resistance and induction of MAPK/JNK activation in head and neck cancer cells. / Ozawa, Hiroyuki; Ranaweera, Ruchira S.; Izumchenko, Evgeny; Makarev, Eugene; Zhavoronkov, Alex; Fertig, Elana J.; Howard, Jason D.; Markovic, Ana; Bedi, Atul; Ravi, Rajani; Perez, Jimena; Le, Quynh Thu; Kong, Christina S.; Jordan, Richard C.; Wang, Hao; Kang, Hyunseok; Quon, Harry; Sidransky, David; Chung, Christine H.

In: Clinical Cancer Research, Vol. 23, No. 17, 01.09.2017, p. 5162-5175.

Research output: Contribution to journalArticle

Ozawa, H, Ranaweera, RS, Izumchenko, E, Makarev, E, Zhavoronkov, A, Fertig, EJ, Howard, JD, Markovic, A, Bedi, A, Ravi, R, Perez, J, Le, QT, Kong, CS, Jordan, RC, Wang, H, Kang, H, Quon, H, Sidransky, D & Chung, CH 2017, 'SMAD4 loss is associated with cetuximab resistance and induction of MAPK/JNK activation in head and neck cancer cells', Clinical Cancer Research, vol. 23, no. 17, pp. 5162-5175. https://doi.org/10.1158/1078-0432.CCR-16-1686
Ozawa, Hiroyuki ; Ranaweera, Ruchira S. ; Izumchenko, Evgeny ; Makarev, Eugene ; Zhavoronkov, Alex ; Fertig, Elana J. ; Howard, Jason D. ; Markovic, Ana ; Bedi, Atul ; Ravi, Rajani ; Perez, Jimena ; Le, Quynh Thu ; Kong, Christina S. ; Jordan, Richard C. ; Wang, Hao ; Kang, Hyunseok ; Quon, Harry ; Sidransky, David ; Chung, Christine H. / SMAD4 loss is associated with cetuximab resistance and induction of MAPK/JNK activation in head and neck cancer cells. In: Clinical Cancer Research. 2017 ; Vol. 23, No. 17. pp. 5162-5175.
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abstract = "Purpose: We previously demonstrated an association between decreased SMAD4 expression and cetuximab resistance in head and neck squamous cell carcinoma (HNSCC). The purpose of this study was to further elucidate the clinical relevance of SMAD4 loss in HNSCC. Experimental Design: SMAD4 expression was assessed by IHC in 130 newly diagnosed and 43 patients with recurrent HNSCC. Correlative statistical analysis with clinicopathologic data was also performed. OncoFinder, a bioinformatics tool, was used to analyze molecular signaling in TCGA tumors with low or high SMAD4 mRNA levels. The role of SMAD4 was investigated by shRNA knockdown and gene reconstitution of HPV-negative HNSCC cell lines in vitro and in vivo. Results: Our analysis revealed that SMAD4 loss was associated with an aggressive, HPV-negative, cetuximab-resistant phenotype. We found a signature of prosurvival and antiapoptotic pathways that were commonly dysregulated in SMAD4-low cases derived from TCGA-HNSCC dataset and an independent oral cavity squamous cell carcinoma (OSCC) cohort obtained from GEO. We show that SMAD4 depletion in an HNSCC cell line induces cetuximab resistance and results in worse survival in an orthotopic mouse model in vivo. We implicate JNK and MAPK activation as mediators of cetuximab resistance and provide the foundation for the concomitant EGFR and JNK/MAPK inhibition as a potential strategy for overcoming cetuximab resistance in HNSCCs with SMAD4 loss. Conclusions: Our study demonstrates that loss of SMAD4 expression is a signature characterizing the cetuximab-resistant phenotype and suggests that SMAD4 expression may be a determinant of sensitivity/resistance to EGFR/MAPK or EGFR/JNK inhibition in HPV-negative HNSCC tumors.",
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T1 - SMAD4 loss is associated with cetuximab resistance and induction of MAPK/JNK activation in head and neck cancer cells

AU - Ozawa, Hiroyuki

AU - Ranaweera, Ruchira S.

AU - Izumchenko, Evgeny

AU - Makarev, Eugene

AU - Zhavoronkov, Alex

AU - Fertig, Elana J.

AU - Howard, Jason D.

AU - Markovic, Ana

AU - Bedi, Atul

AU - Ravi, Rajani

AU - Perez, Jimena

AU - Le, Quynh Thu

AU - Kong, Christina S.

AU - Jordan, Richard C.

AU - Wang, Hao

AU - Kang, Hyunseok

AU - Quon, Harry

AU - Sidransky, David

AU - Chung, Christine H.

PY - 2017/9/1

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N2 - Purpose: We previously demonstrated an association between decreased SMAD4 expression and cetuximab resistance in head and neck squamous cell carcinoma (HNSCC). The purpose of this study was to further elucidate the clinical relevance of SMAD4 loss in HNSCC. Experimental Design: SMAD4 expression was assessed by IHC in 130 newly diagnosed and 43 patients with recurrent HNSCC. Correlative statistical analysis with clinicopathologic data was also performed. OncoFinder, a bioinformatics tool, was used to analyze molecular signaling in TCGA tumors with low or high SMAD4 mRNA levels. The role of SMAD4 was investigated by shRNA knockdown and gene reconstitution of HPV-negative HNSCC cell lines in vitro and in vivo. Results: Our analysis revealed that SMAD4 loss was associated with an aggressive, HPV-negative, cetuximab-resistant phenotype. We found a signature of prosurvival and antiapoptotic pathways that were commonly dysregulated in SMAD4-low cases derived from TCGA-HNSCC dataset and an independent oral cavity squamous cell carcinoma (OSCC) cohort obtained from GEO. We show that SMAD4 depletion in an HNSCC cell line induces cetuximab resistance and results in worse survival in an orthotopic mouse model in vivo. We implicate JNK and MAPK activation as mediators of cetuximab resistance and provide the foundation for the concomitant EGFR and JNK/MAPK inhibition as a potential strategy for overcoming cetuximab resistance in HNSCCs with SMAD4 loss. Conclusions: Our study demonstrates that loss of SMAD4 expression is a signature characterizing the cetuximab-resistant phenotype and suggests that SMAD4 expression may be a determinant of sensitivity/resistance to EGFR/MAPK or EGFR/JNK inhibition in HPV-negative HNSCC tumors.

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