Genome-wide epigenetic and mRNA-expression profiling followed by CRISPR/Cas9-mediated gene-disruptions corroborate the MIR141/MIR200C -ZEB1/ZEB2-FGFR1 axis in acquired EMT-associated EGFR TKI-resistance in NSCLC cells.

Autor:	Vad-Nielsen J; Department of Biomedicine, Aarhus University, Aarhus, Denmark., Staunstrup NH; Department of Biomedicine, Aarhus University, Aarhus, Denmark., Kjeldsen ML; Department of Biomedicine, Aarhus University, Aarhus, Denmark., Dybdal N; Department of Biomedicine, Aarhus University, Aarhus, Denmark., Flandin G; Department of Biomedicine, Aarhus University, Aarhus, Denmark., De Stradis C; Department of Biomedicine, Aarhus University, Aarhus, Denmark., Daugaard TF; Department of Biomedicine, Aarhus University, Aarhus, Denmark., Vilsbøll-Larsen T; Department of Biomedicine, Aarhus University, Aarhus, Denmark., Maansson CT; Department of Biomedicine, Aarhus University, Aarhus, Denmark.; Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark.; Department of Clinical Medicine, Aarhus University, Aarhus, Denmark., Doktor TK; Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M, Denmark., Sorensen BS; Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark.; Department of Clinical Medicine, Aarhus University, Aarhus, Denmark., Nielsen AL; Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Jazyk:	angličtina
Zdroj:	Translational lung cancer research [Transl Lung Cancer Res] 2023 Jan 31; Vol. 12 (1), pp. 42-65. Date of Electronic Publication: 2023 Jan 13.
DOI:	10.21037/tlcr-22-507
Abstrakt:	Background: Epithelial-mesenchymal-transition (EMT) is an epigenetic-based mechanism contributing to the acquired treatment resistance against receptor tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC) cells harboring epidermal growth factor receptor ( EGFR )-mutations. Delineating the exact epigenetic and gene-expression alterations in EMT-associated EGFR TKI-resistance (EMT-E-TKI-R) is vital for improved diagnosis and treatment of NSCLC patients. Methods: We characterized genome-wide changes in mRNA-expression, DNA-methylation and the histone-modification H3K36me3 in EGFR -mutated NSCLC HCC827 cells in result of acquired EMT-E-TKI-R. CRISPR/Cas9 was used to functional examine key findings from the omics analyses. Results: Acquired EMT-E-TKI-R was analyzed with three omics approaches. RNA-sequencing identified 2,233 and 1,972 up- and down-regulated genes, respectively, and among these were established EMT-markers. DNA-methylation EPIC array analyses identified 14,163 and 7,999 hyper- and hypo-methylated, respectively, differential methylated positions of which several were present in EMT-markers. Finally, H3K36me3 chromatin immunoprecipitation (ChIP)-sequencing detected 2,873 and 3,836 genes with enrichment and depletion, respectively, and among these were established EMT-markers. Correlation analyses showed that EMT-E-TKI-R mRNA-expression changes correlated better with H3K36me3 changes than with DNA-methylation changes. Moreover, the omics data supported the involvement of the MIR141/MIR200C -ZEB1/ZEB2-FGFR1 signaling axis for acquired EMT-E-TKI-R. CRISPR/Cas9-mediated analyses corroborated the importance of ZEB1 in acquired EMT-E-TKI-R, MIR200C and MIR141 to be in an EMT-E-TKI-R-associated auto-regulatory loop with ZEB1, and FGFR1 to mediate cell survival in EMT-E-TKI-R. Conclusions: The current study describes the synchronous genome-wide changes in mRNA-expression, DNA-methylation, and H3K36me3 in NSCLC EMT-E-TKI-R. The omics approaches revealed potential novel diagnostic markers and treatment targets. Besides, the study consolidates the functional impact of the MIR141/MIR200C -ZEB1/ZEB2-FGFR1-signaling axis in NSCLC EMT-E-TKI-R. Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://tlcr.amegroups.com/article/view/10.21037/tlcr-22-507/coif). The authors have no conflicts of interest to declare. (2023 Translational Lung Cancer Research. All rights reserved.)
Databáze:	MEDLINE
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