The NF-κB/ALDH1A1 signaling is established as a key non-genetic mechanism of acquired EGFR-TKI resistance and a rational combination strategy to overcome it is provided.
Abstract
Acquired resistance to tyrosine kinase inhibitors (TKIs) remains a major clinical challenge in the treatment of EGFR-mutant non-small cell lung cancer (NSCLC). This study established TKI-resistant variants by integrating cell lines, lung cancer organoids (LCOs), and in vivo models, revealing the pivotal role of the NF-κB/ALDH1A1 signaling in mediating non-mutational TKI resistance. Resistant cells exhibited elevated RELA phosphorylation, enhanced ALDH1A1 expression and enzymatic activity, and stem-like properties. Mechanistically, NF-κB activation occurred as an early response to TKI exposure and promoted ALDH1A1 transcription via RELA. In turn, ALDH1A1 contributed to the sustained activation of NF-κB signaling, forming a self-reinforcing positive feedback loop. Genetic ALDH1A1 or RELA silencing reversed the resistant phenotype. Pharmacologically, treatment with an EGFR-TKI and the ALDH1A1 inhibitor disulfiram or the NF-κB-targeting agent EGCG synergistically restored the antitumor efficacy of TKIs both in vitro and in vivo. These findings establish the NF-κB/ALDH1A1 signaling as a key non-genetic mechanism of acquired EGFR-TKI resistance and provide a rational combination strategy to overcome it. Catalysis-dependent NF-κB–ALDH1A1 feedback loop drives non-mutational EGFR-TKI resistance. EGFR-TKI treatment suppresses EGFR signaling but rapidly induces NF-κB activation as an early event under TKI stress in EGFR-mutant NSCLC cells. Activated NF-κB (p-RELA) drives ALDH1A1 transcription, while ALDH1A1 enzymatic activity reinforces NF-κB signaling via IKKβ phosphorylation, forming a catalysis-dependent positive feedback loop that sustains stemness and promotes non-mutational TKI resistance. Disruption of this axis by inhibiting ALDH1A1 (DSF) or NF-κB (EGCG) restores TKI sensitivity. Catalysis-dependent NF-κB–ALDH1A1 feedback loop drives non-mutational EGFR-TKI resistance. EGFR-TKI treatment suppresses EGFR signaling but rapidly induces NF-κB activation as an early event under TKI stress in EGFR-mutant NSCLC cells. Activated NF-κB (p-RELA) drives ALDH1A1 transcription, while ALDH1A1 enzymatic activity reinforces NF-κB signaling via IKKβ phosphorylation, forming a catalysis-dependent positive feedback loop that sustains stemness and promotes non-mutational TKI resistance. Disruption of this axis by inhibiting ALDH1A1 (DSF) or NF-κB (EGCG) restores TKI sensitivity.
Resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) represents a major clinical challenge in the management of non-small cell lung cancer (NSCLC). Chromosome 4q12 locus is an important gene locus associated with progression-free survival (PFS) in NSCLC patients receiving EGFR-TKIs therapy. However, it remains poorly characterized how genes at this locus function in NSCLC development and resistance to EGFR-TKIs. Here, we found that CLOCK at this locus is highly expressed in NSCLC tissues and correlates with unfavorable PFS of patients. CLOCK promotes malignant proliferation and metastasis of NSCLC in vitro and in vivo. CLOCK could attenuate treatment efficacies of gefitinib (one of the first-generation EGFR-TKIs) or osimertinib (one of the third-generation EGFR-TKIs). Mechanistically, CLOCK functions as a transcriptional factor to upregulate LAMC2 transcription and expression in NSCLC cells. There was significantly elevated LAMC2 expressed in NSCLC tissues and its high levels were associated with shortened survival of patients. Indeed, CLOCK could activate multiple kinase signaling pathways, such as the PI3K-Akt signaling and the MAPK signaling, by facilitating either the LAMC2-ITGB1 interaction and the LAMC2-EGFR interaction and thereby accelerating NSCLC proliferation and conferring EGFR-TKI resistance. Collectively, these findings for the first time identify CLOCK as a critical mediator of EGFR-TKIs resistance and a promising target to overcome EGFR-TKIs resistance in NSCLC.
Yankang Li, Linyu Han, Huiwen Luan et al.· Archives of Biochemistry and...· 0 citations
INTRODUCTION
Tumor immune evasion is a pivotal mechanism driving therapeutic resistance and poor prognosis in lung cancer. The interleukin-20 receptor β subunit (IL20RB) is implicated in chronic inflammation and oncogenesis, but its precise function and molecular basis in non-small cell lung cancer (NSCLC) require elucidation.
METHODS
Multi-omics data analysis, clinical NSCLC specimens, and a series of in vitro and in vivo functional experiments were employed to investigate the role of IL20RB. Techniques included gene expression correlation, survival analysis, cell proliferation/migration/invasion assays, co-immunoprecipitation, western blotting, and pharmacologic modulation studies.
RESULTS
IL20RB was significantly overexpressed in NSCLC tissues, correlating with advanced disease stage and inferior patient survival. Its knockdown potently suppressed NSCLC cell proliferation, migration, and invasion. High IL20RB expression was associated with reduced T cell infiltration and function within the tumor microenvironment. Mechanistically, IL20RB directly bound to JAK2 via a critical lysine residue (K267), activating JAK2-STAT3 signaling and upregulating downstream effectors including PD-L1 and BCL2, thereby coordinating tumor growth and immune escape. The commonly used drug acetaminophen (APAP) was identified to downregulate IL20RB expression via HDAC3-mediated deacetylation, inhibiting this oncogenic pathway and demonstrating synergistic activity with anti-PD-L1 therapy in vitro.
CONCLUSIONS
IL20RB drives NSCLC progression and immune evasion through a K267-dependent activation of the JAK2-STAT3 pathway. Pharmacologic suppression of IL20RB by APAP presents a novel therapeutic strategy, positioning IL20RB as a promising prognostic biomarker and therapeutic target in NSCLC.
Yanghao Wang, Guo-Yun Li, Wei-Zhou Wang et al.· Lung Cancer· 0 citations
A key role is found for NF-κB/Lipocalin 2 (LCN2)-signaling-pathway-induced ferroptosis in modulating cisplatin sensitivity of V600E-overexpressing CRC cells to cisplatin within a KRAS-mutant cellular background, implying LCN2 as a potential therapeutic target.
M. Feng, Xuesong Wu, Li Jiang et al.· Cancers· 0 citations
Target neutralization of SAA1 synergized robustly with osimertinib to reverse immunosuppressive remodeling and induce profound tumor regression in multiple preclinical models, highlighting SAA1 neutralization as a tractable combinatorial strategy to enhance EGFR-TKI efficacy in LUAD.
Jun-Kan Zhu, Shencheng Ren, Tao Cheng et al.· Journal of Experimental &...· 0 citations
Background Non‑small cell lung cancer (NSCLC) is a major cause of cancer‑associated death globally. Elucidating novel molecular drivers is essential for targeted therapy development. This study sought to investigate the function and regulatory mechanism of proteasome 20S subunit alpha 6 (PSMA6) in NSCLC. Methods Proteasome 20S subunit alpha 6 (PSMA6) expression in NSCLC tissues and cells was analyzed using The Cancer Genome Atlas (TCGA) datasets, immunohistochemistry, and Western blotting. Gain- and loss-of-function assays were performed to evaluate its biological functions. Protein interaction assays and functional rescue experiments were conducted to investigate the underlying mechanisms. Results We found that PSMA6 was significantly upregulated in NSCLC tissues and cell lines and was associated with poor patient prognosis. Functional experiments demonstrated that PSMA6 promoted NSCLC cell proliferation and migration. Mechanistically, PSMA6 activated nuclear factor kappa B (NF-κB) signaling by enhancing p65 nuclear translocation and IκBα phosphorylation. Further investigation revealed that PSMA6 directly interacted with neural precursor cell expressed, developmentally down-regulated 8 (NEDD8) and increased its protein stability without affecting its mRNA level. Importantly, NEDD8 knockdown abolished PSMA6-induced NF-κB activation and malignant phenotypes, confirming that PSMA6 exerts its oncogenic effects through the NEDD8/NF-κB axis. Conclusions Our findings identify a novel PSMA6/NEDD8/NF-κB regulatory axis that promotes NSCLC progression and suggest PSMA6 as a potential therapeutic target.
Shi-Yang Deng, Zi-Xuan Li, Nan-Xi Liu et al.· Translational Lung Cancer Re...· 0 citations
PURPOSE
Aberrant activation of insulin-like growth factor 1 receptor (IGF1R) in hepatocellular carcinoma (HCC) is closely associated with resistance to targeted therapies. However, the mechanism by which IGF1R transcriptionally upregulates programmed death-ligand 1 (PD-L1) to drive sorafenib resistance remained unclear. This study aimed to elucidate the molecular pathway through which IGF1R upregulates PD-L1 via the PI3K/AKT/NF-κB signaling cascade, thereby promoting sorafenib resistance.
METHODS
The regulatory network of the IGF1R-PI3K/AKT/NF-κB-PD-L1 axis was analyzed using immunohistochemistry (20 HCC clinical samples), Western blotting (THLE-2 and Huh7/Hep3B/SNU-387 cells), functional assays (CCK8, colony formation, Transwell, JC-1, Annexin V-FITC/PI), chromatin immunoprecipitation (ChIP), and dual-luciferase reporter assays. Two independent in vivo cohorts were established: Cohort 1(SCID mice bearing Huh7 xenografts, IGF1 + sorafenib (SFB)) to assess IGF1R-driven resistance; Cohort 2 (NOD-SCID mice/SNU-387, PD-L1 knockdown/overexpression ± sorafenib) to validate PD-L1 as the downstream effector.
RESULTS
IGF1R was significantly overexpressed in HCC. IGF1R activation increased the sorafenib IC50 by 2.3-fold and activated PI3K/AKT/NF-κB signaling. NF-κB directly bound to the PD-L1 promoter to drive its transcription, as confirmed by ChIP and dual-luciferase assays. PI3K inhibition with LY294002 blocked IGF1R-induced PD-L1 upregulation and restored sorafenib sensitivity. PD-L1 overexpression promoted HCC proliferation, migration, and cell-cycle progression while suppressing apoptosis; PD-L1 knockdown yielded opposite effects. In vivo, Cohort 1 confirmed that IGF1R activation promoted tumor growth, attenuated sorafenib efficacy, and elevated p-AKT, p-NF-κB, and PD-L1 in tumor tissues. Cohort 2 demonstrated that PD-L1 knockdown augmented sorafenib-induced tumor suppression, whereas PD-L1 overexpression blunted sorafenib efficacy, with reciprocal changes in Ki-67 and pathway phosphorylation.
CONCLUSION
IGF1R upregulates PD-L1 via the PI3K/AKT/NF-κB axis through direct NF-κB-mediated transcriptional activation, and PD-L1 serves as a critical downstream effector driving sorafenib resistance in HCC.
Tao Zhu, Chunmei Xie, Qianqian Shao et al.· Molecular Immunology· 0 citations
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