Failure to EGFR-TKI-based therapy and tumoural progression are promoted by MEOX2/GLI1-mediated epigenetic regulation of EGFR in the human lung cancer

• Published in: European journal of cancer (1990) 2022-01, Vol.160, p.189-205
• Main Authors: Peralta-Arrieta, Irlanda, Trejo-Villegas, Octavio A., Armas-López, Leonel, Ceja-Rangel, Hugo A., Ordóñez-Luna, María del Carmen, Pineda-Villegas, Priscila, González-López, Marco A., Ortiz-Quintero, Blanca, Mendoza-Milla, Criselda, Zatarain-Barrón, Zyanya L., Arrieta, Oscar, Zúñiga, Joaquín, Ávila-Moreno, Federico
• Format: Article
• Language: English
• Subjects: AKT protein; Antineoplastic Agents - pharmacology; Antineoplastic Agents - therapeutic use; Assaying; Biomarkers; Cancer drug resistance; Cell Line, Tumor; Cell proliferation; Chemoresistance; Cisplatin; Cytotoxicity; Disease Progression; EGFR-TKIs; Epidermal growth factor; Epidermal growth factor receptors; Epigenesis, Genetic - genetics; Epigenetics; Erlotinib; Extracellular signal-regulated kinase; Gene expression; Gene sequencing; Genes, erbB-1 - drug effects; GLI1; Glioma; Growth factors; Histones; Homeobox; Humans; Kinases; Lung cancer; Lung Neoplasms - drug therapy; Malignancy; MEOX2; Mesenchyme; Osimertinib; Polycomb group proteins; Protein Kinase Inhibitors - therapeutic use; Protein-tyrosine kinase; Resistance factors; Signal transduction; Therapy; Transcription factors; Tumors; Tyrosine; Zinc Finger Protein GLI1 - genetics
• ISSN: 0959-8049; 1879-0852
• DOI: 10.1016/j.ejca.2021.10.032

Mesenchyme homeobox-2 (MEOX2)-mediated regulation of glioma-associated oncogene-1 (GLI1) has been associated with poor overall survival, conferring chemoresistance in lung cancer. However, the role of MEOX2/GLI1 in resistance to epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs)-based therapy remains unexplored in human lung cancer. Functional assays using genetic silencing strategy by short hairpin RNAs, as well as cytotoxic (tetrazolium dye MTT) and clonogenic assays, were performed to evaluate MEOX2/GLI1-induced malignancy capacity in lung cancer cells. Further analysis performed includes western blot, qPCR and ChIP-qPCR assays to identify whether MEOX2/GLI1 promote EGFR/AKT/ERK activation, as well as EGFR overexpression through epigenetic mechanisms. Finally, preclinical tumour progression in vivo and progression-free disease interval analyses in patients treated with EGFR-TKI were included. Overexpressed MEOX2/GLI1 in both EGFR wild-type and EGFR/KRAS-mutated lung cancer cells were detected and involved in the activation/expression of EGFR/AKT/ERK biomarkers. In addition, MEOX2/GLI1 was shown to be involved in the increased proliferation of tumour cells and resistance capacity to cisplatin, EGFR-TKIs (erlotinib and AZD9291 ‘osimertinib’), AZD8542-SMO, and AZD6244-MEKK1/2. In addition, we identified that MEOX2/GLI1 promote lung tumour cells progression in vivo and are clinically associated with poorer progression-free disease intervals. Finally, both MEOX2 and GLI1 were detected to be epigenetically involved in EGFR expression by reducing both repressive markers polycomb-EZH2 and histone H3K27me3, but, particularly, increasing an activated histone profile H3K27Ac/H3K4me3 at EGFR-gene enhancer-promoter sequences that probably representing a novel EGFR-TKI-based therapy resistance mechanism. MEOX2/GLI1 promote resistance to cisplatin and EGFR-TKI-based therapy in lung cancer cells, modulating EGFR/AKT/ERK signalling pathway activation, as well as inducing an aberrant epigenetic modulation of the EGFR-gene expression in human lung cancer. [Display omitted] •Resistance to EGFR-TKI based treatment occurs through MEOX2/GLI1 transcription factors in lung cancer cells.•Overexpressed MEOX2/GLI1 promotes cisplatin resistance and lung tumour cells proliferation.•MEOX2/GLI promotes in vitro resistance to EGFR-TKI therapy by EGFR-pathway over-activation.•Overexpressed MEOX2/GLI1 increase in vivo lung tumour cells progression.•MEOX2/GLI1 modulat