TNF-driven adaptive response mediates resistance to EGFR inhibition in lung cancer
Ke Gong, … , Dawen Zhao, Amyn A. Habib
Ke Gong, … , Dawen Zhao, Amyn A. Habib
Published April 3, 2018
Citation Information: J Clin Invest. 2018;128(6):2500-2518. https://doi.org/10.1172/JCI96148.
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Research Article

TNF-driven adaptive response mediates resistance to EGFR inhibition in lung cancer

Abstract

Although aberrant EGFR signaling is widespread in cancer, EGFR inhibition is effective only in a subset of non–small cell lung cancer (NSCLC) with EGFR activating mutations. A majority of NSCLCs express EGFR wild type (EGFRwt) and do not respond to EGFR inhibition. TNF is a major mediator of inflammation-induced cancer. We find that a rapid increase in TNF level is a universal adaptive response to EGFR inhibition in NSCLC, regardless of EGFR status. EGFR signaling actively suppresses TNF mRNA levels by inducing expression of miR-21, resulting in decreased TNF mRNA stability. Conversely, EGFR inhibition results in loss of miR-21 and increased TNF mRNA stability. In addition, TNF-induced NF-κB activation leads to increased TNF transcription in a feed-forward loop. Inhibition of TNF signaling renders EGFRwt-expressing NSCLC cell lines and an EGFRwt patient-derived xenograft (PDX) model highly sensitive to EGFR inhibition. In EGFR-mutant oncogene-addicted cells, blocking TNF enhances the effectiveness of EGFR inhibition. EGFR plus TNF inhibition is also effective in NSCLC with acquired resistance to EGFR inhibition. We suggest concomitant EGFR and TNF inhibition as a potentially new treatment approach that could be beneficial for a majority of lung cancer patients.

Authors

Ke Gong, Gao Guo, David E. Gerber, Boning Gao, Michael Peyton, Chun Huang, John D. Minna, Kimmo J. Hatanpaa, Kemp Kernstine, Ling Cai, Yang Xie, Hong Zhu, Farjana J. Fattah, Shanrong Zhang, Masaya Takahashi, Bipasha Mukherjee, Sandeep Burma, Jonathan Dowell, Kathryn Dao, Vassiliki A. Papadimitrakopoulou, Victor Olivas, Trever G. Bivona, Dawen Zhao, Amyn A. Habib

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Figure 9

EGFR and TNF inhibition prevents the acquired resistance.

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EGFR and TNF inhibition prevents the acquired resistance. (A) TetO-EGFR-...
(A) TetO-EGFR-L858R and CCSP-rtTA mice were exposed to doxycycline diets to induce tumors. Lung tumor formation was confirmed by MRI between weeks 4 and 5. Starting from week 5, mice were randomly divided into 4 groups and treated by 6.25 mg/kg erlotinib and/or 150 mg/kg thalidomide for 2 weeks. Tumor sizes were measured by 2 blinded researchers using ImageJ. Data were presented as each tumor size and the mean ± SEM. (B) MRI images from representative mice in each group. The tumors grow as diffuse lung opacities. (C) TNF mRNAs were detected in HCC827 parent and erlotinib-resistant cell lines by qPCR. (D–F) HCC827/ER3, HCC827/ER4A, and H1975 cells were treated with 5 μg/ml thalidomide and/or 100 nM erlotinib/afatinib for 72 hours followed by AlamarBlue assay. (G) 1 × 106 H1975 cells were injected s.c. into athymic mice. When tumors formed, mice were divided into 4 groups (n = 6) and treated with 5 mg/kg afatinib by oral gavage or 150 mg/kg thalidomide i.p. for 24 days. (H) HCC827 cells were planted in a 96-well plate and treated with 100 nM erlotinib with or without 5 μg/ml thalidomide. The day when cells reached 100% confluence was considered the appearance of acquired resistance. (I) HCC827 cells were injected subcutaneously into athymic mice. When tumors reached 500 mm3, mice were divided into 4 groups (n = 6) and treated with 6.25 mg/kg erlotinib by oral gavage or 150 mg/kg thalidomide by i.p. for 32 days. For (C–F) data represent mean ± SEM. n = 3 biologically independent experimental replicates. *P < 0.05, **P < 0.01, ***P < 0.001, by 2-sample t-test, except H by log-rank test. For G and I, statistical significance was defined as P < 0.05 (repeated-measures 2-way ANOVA, Bonferroni correction, GraphPad Prism 7.0).