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Bispecific T-cell Engager (CD3 × EGFR)-Based Triplet Therapy Unlocks CD40/CD40L Crosstalk to Revert Immunosuppression in Third-Generation EGFR-Tyrosine Kinase Inhibitor-Refractory NSCLC.

Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer 2026 p. 103607

Guo H, Wang Y, Diao M, Han R, He T, Wu Y, Wang Q, Cheng L, Zhao C, Li X, Xiong A, Li W, Zhou F, Zhou C

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[INTRODUCTION] Acquired resistance to third-generation EGFR-tyrosine kinase inhibitors (EGFR-TKIs) is a major challenge in NSCLC, with approximately 50% of cases lacking precise resistance mechanisms.

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  • p-value p < 0.001

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APA Guo H, Wang Y, et al. (2026). Bispecific T-cell Engager (CD3 × EGFR)-Based Triplet Therapy Unlocks CD40/CD40L Crosstalk to Revert Immunosuppression in Third-Generation EGFR-Tyrosine Kinase Inhibitor-Refractory NSCLC.. Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 103607. https://doi.org/10.1016/j.jtho.2026.103607
MLA Guo H, et al.. "Bispecific T-cell Engager (CD3 × EGFR)-Based Triplet Therapy Unlocks CD40/CD40L Crosstalk to Revert Immunosuppression in Third-Generation EGFR-Tyrosine Kinase Inhibitor-Refractory NSCLC.." Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 2026, pp. 103607.
PMID 41672197

Abstract

[INTRODUCTION] Acquired resistance to third-generation EGFR-tyrosine kinase inhibitors (EGFR-TKIs) is a major challenge in NSCLC, with approximately 50% of cases lacking precise resistance mechanisms. This study investigates the immunosuppressive tumor microenvironment (TME) driving resistance and develops a novel triple-combination therapy to restore T-cell antitumor activity.

[METHODS] Single-cell RNA sequencing and multicolor fluorescence staining were performed on NSCLC patient samples to analyze TME changes post-EGFR-TKI resistance. A triple therapy combining BC3448 (EGFR and CD3 bispecific T-cell engager), tucidinostat (histone deacetylase inhibitor), and WBP3425 (4-1BB agonist) was tested using in vitro co-culture assays, syngeneic cell-derived xenograft models in humanized NOG-EXL mice, and multi-omics analyses.

[RESULTS] Single-cell RNA sequencing revealed reduced T-cell infiltration or activation and increased immunosuppressive myeloid cells in resistant NSCLC. BC3448 monotherapy activated T cells and induced tumor cell apoptosis in vitro but was limited in vivo because of myeloid-driven immunosuppression. The triple therapy significantly enhanced tumor regression in osimertinib-resistant models (tumor growth inhibition >70%, p < 0.001), promoted CD8+ effector T-cell differentiation, and suppressed Tregs and M2 macrophages. CD40-CD40L axis activation between T cells and monocyte-derived macrophages was critical for TME remodeling, with spatial profiling revealing increased CD40L+ T-cell and CD40+ macrophage proximity, correlating with higher IFN-γ and reduced angiogenesis. A durable response to BC3448 monotherapy was observed in an immunotherapy-resistant patient with NSCLC (>2 y of stable disease), presenting a translational potential of this approach.

[CONCLUSIONS] This study establishes a novel triple therapy that overcomes the limitations of bispecific T-cell engagers in cold and immunosuppressive TMEs and provides an immunomodulatory approach to addressing third-generation EGFR-TKI resistance.

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