EGFR activation disrupts immunotherapy response via SHP2-mediated suppression of tumor-intrinsic response to IFN-γ
Wei-Tao Zhuang, Lan-Lan Pang, Li-Yang Hu, Jun Liao, Jian-Hua Zhan, Ting Li, Ri-Xin Chen, Jia-Ni Zheng, An-Lin Li, Wen-Yan Yu, Tian-Qin Mao, Liang Chen, Yu-Jian Huang, Shao-Dong Hong, Jing Li, Jun-Han Wu, Yi-Ming Zeng, Meng-Juan Yang, Hai-Qing Zeng, Ya-Xiong Zhang, Li Zhang, Wen-Feng Fang
Wei-Tao Zhuang, Lan-Lan Pang, Li-Yang Hu, Jun Liao, Jian-Hua Zhan, Ting Li, Ri-Xin Chen, Jia-Ni Zheng, An-Lin Li, Wen-Yan Yu, Tian-Qin Mao, Liang Chen, Yu-Jian Huang, Shao-Dong Hong, Jing Li, Jun-Han Wu, Yi-Ming Zeng, Meng-Juan Yang, Hai-Qing Zeng, Ya-Xiong Zhang, Li Zhang, Wen-Feng Fang
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Research Article Immunology Oncology

EGFR activation disrupts immunotherapy response via SHP2-mediated suppression of tumor-intrinsic response to IFN-γ

Abstract

Epidermal growth factor receptor–activating (EGFR-activating) mutations are established biomarkers of resistance to immune checkpoint blockade (ICB) in lung cancer, yet the precise molecular mechanism and effective therapeutic strategies remain elusive. In this study, we show that EGFR overexpression and amplification recapitulated the negative effect of EGFR driver mutations on the ICB response, indicating a proactive involvement of EGFR signaling in antagonizing the antitumor immune response. Functional studies unveiled that EGFR activation suppressed the cellular response to IFN-γ following ICB treatment across multiple cancer models. This impairment in IFN-γ responsiveness further limited the upregulation of T cell–recruiting chemokines and antigen presentation, resulting in reduced T cell infiltration and activation, ultimately undermining antitumor immunity. Mechanistically, EGFR promotes Src homology 2-containing protein tyrosine phosphatase 2 (SHP2) activation to accelerate STAT1 dephosphorylation, leading to premature termination of the IFN-γ response. SHP2 inhibition restored ICB sensitivity in EGFR-activated tumors, significantly reducing tumor burden while maintaining a favorable safety profile. Our findings suggest that the EGFR/SHP2 axis functions as a molecular brake to disrupt the initiation and amplification of the IFN-γ–mediated antitumor response during immunotherapy. This discovery unveils a potential avenue to overcome immunotherapy resistance in EGFR-driven tumors, particularly lung cancer, through SHP2-targeted combination strategies.

Authors

Wei-Tao Zhuang, Lan-Lan Pang, Li-Yang Hu, Jun Liao, Jian-Hua Zhan, Ting Li, Ri-Xin Chen, Jia-Ni Zheng, An-Lin Li, Wen-Yan Yu, Tian-Qin Mao, Liang Chen, Yu-Jian Huang, Shao-Dong Hong, Jing Li, Jun-Han Wu, Yi-Ming Zeng, Meng-Juan Yang, Hai-Qing Zeng, Ya-Xiong Zhang, Li Zhang, Wen-Feng Fang

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Figure 6

EGFR promotes SHP2 activation to accelerate STAT1 dephosphorylation and abort the IFN-γ response.

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EGFR promotes SHP2 activation to accelerate STAT1 dephosphorylation and ...
(A and B) Western blot analysis of IFNGR1 and IFNGR2 expression in WT EGFR and EGFR-mutant cell lines (A), quantified in ImageJ by pixel intensities and normalized to GAPDH (B). (C) RNA expression of IFNGR1 and IFNGR2 in WT EGFR and EGFR-mutant LUAD from TCGA dataset. (D and E) Western blot analysis of IFN-γ signaling markers (pJAK2, pSTAT1, IRF1) in EGFR-WT and EGFR-mutant cell lines with or without IFN-γ stimulation (10 ng/mL) (D), quantified in ImageJ by pixel intensities and normalized to GAPDH (E). (F) Western blot analysis of pSTAT1 (Tyr701) at baseline or following stimulation with IFN-γ (10 ng/mL) for 24 hours in H1299, Beas-2B, or A549 cells stably expressing EV, EGFRL858R, or EGFR19del. (G) Schematic illustration of dephosphorylation assays of pSTAT1 (Tyr701). (H and I) Western blot analysis of pSTAT1Tyr701 at the indicated time points following IFN-γ exposure in H1299-EV/EGFRL858R cells (H), as outlined in G. Protein levels were quantified in ImageJ and then plotted as trend curves (I). See also Supplemental Figure 6, C and D. (J) Identification of the amino acid sequence of SHP2 by mass spectrometric analysis of the IP pulldown. (K) Western blot analysis of pSHP2Tyr542 and downstream pERK1/2 in H1299-EV/EGFRL858R cell models treated with a concentration gradient of EGF (0, 1, 10, 50, 100, 500 ng/mL) or osimertinib (0, 10, 50, 100, 1,000, 10,000 nM) for 3 hours. (L) FACS analysis of mCherry fluorescence in H1299-EGFRL858R cells pretreated with a gradient of RMC4550 or SHP099 for 3 hours, followed by combination with IFN-γ (5 ng/mL) for 24 hours. Data are presented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, by unpaired, 2-tailed t test (B), Wilcoxon rank-sum test (C), 2-way ANOVA (E), or 1-way ANOVA with Tukey’s test (L).