Dataset Information

Adaptive resistance to Afatinib in non-small cell lung cancer cells in attenuated by Ca2+ depletion

ABSTRACT: Targeted therapies against oncogenic receptor tyrosine kinases are showing promising results in the clinic. However, despite the initial response, most patients become resistant. The majority of research regarding resistance has been focused on acquired resistance occurring after an extensive time of treatment. This has resulted in the development of combination treatments. Yet, the question remains how cells are able to survive initial treatment. We hypothesize that early resistance to apoptosis will enable cells to develop any growth stimulating mechanism. Our results show that regardless of an initial drug response, drug sensitive cells rapidly adapted to targeted therapy. Within days they continued to proliferate, despite persisting target inhibition. The Non-small Cell Lung Cancer (NSCLC) PC9 cell line was used for analysis of its full proteome, kinome and phospho-proteome during early adaptive resistance development against the EGFR targeted Tyrosine Kinase Inhibitor (TKI) Afatinib. The proteomics data showed an increase in abundance and activity of cytoskeleton and calcium signaling related proteins. In addition, cells showed a distinct morphological change. We found that if cells were deprived from calcium in order to restrict them from reorganizing their cytoskeleton, they were no longer able to survive TKI treatment. This could be validated on four different NSCLC cell lines, bearing different genetic backgrounds and sensitivities to initial TKI treatment targeting the EGFR. Therapeutically, increasing the efficiency of TKI treatment, and therefore early inhibition of adaption could potentially prevent further resistance development.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Non-small Cell Lung Carcinoma

SUBMITTER: Celine Mulder

LAB HEAD: S.M. Lemeer

PROVIDER: PXD008454 | Pride | 2018-10-02

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	201706011_OR13_A4_NP_TCM_PC9_Af_IMAC_2D-1.raw	Raw
	201706011_OR13_A4_NP_TCM_PC9_Af_IMAC_2D-2.raw	Raw
	201706011_OR13_A4_NP_TCM_PC9_Af_IMAC_30min-2.raw	Raw
	201706011_OR13_A4_NP_TCM_PC9_Af_IMAC_5D-1.raw	Raw
	201706011_OR13_A4_NP_TCM_PC9_Af_IMAC_DMSO-2.raw	Raw

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Publications

Adaptive Resistance to EGFR-Targeted Therapy by Calcium Signaling in NSCLC Cells.

Mulder Celine C Prust Nadine N van Doorn Sander S Reinecke Maria M Kuster Bernhard B van Bergen En Henegouwen Paul P Lemeer Simone S

Molecular cancer research : MCR 20180702 11

Targeted therapies against oncogenic receptor tyrosine kinases (RTK) show promising results in the clinic. Unfortunately, despite the initial positive response, most patients develop therapeutic resistance. Most research has focused on acquired resistance occurring after an extensive time of treatment; however, the question remains as to how cells can survive an initial treatment, as early resistance to apoptosis will enable cells to develop any growth-stimulating mechanism. Here, the non-small ...[more]

PMID: 29967110

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Project description:Despite initial and often dramatic responses of epidermal growth factor receptor (EGFR)-addicted lung tumors to the EGFR-specific tyrosine kinase inhibitors (TKIs), gefitinib and erlotinib, nearly all develop resistance and relapse. To explore novel mechanisms mediating acquired resistance, we employed non-small-cell lung cancer (NSCLC) cell lines bearing activating mutations in EGFR and rendered them resistant to EGFR-specific TKIs through chronic adaptation in tissue culture. In addition to previously observed resistance mechanisms including EGFR-T790M 'gate-keeper' mutations and MET amplification, a subset of the seven chronically adapted NSCLC cell lines including HCC4006, HCC2279 and H1650 cells exhibited marked induction of fibroblast growth factor (FGF) 2 and FGF receptor 1 (FGFR1) mRNA and protein. Also, adaptation to EGFR-specific TKIs was accompanied by an epithelial to mesenchymal transition (EMT) as assessed by changes in CDH1, VIM, ZEB1 and ZEB2 expression and altered growth properties in Matrigel. In adapted cell lines exhibiting increased FGF2 and FGFR1 expression, measures of growth and signaling, but not EMT, were blocked by FGFR-specific TKIs, an FGF-ligand trap and FGFR1 silencing with RNAi. In parental HCC4006 cells, cell growth was strongly inhibited by gefitinib, although drug-resistant clones progress within 10 days. Combined treatment with gefitinib and AZD4547, an FGFR-specific TKI, prevented the outgrowth of drug-resistant clones. Thus, induction of FGF2 and FGFR1 following chronic adaptation to EGFR-specific TKIs provides a novel autocrine receptor tyrosine kinase-driven bypass pathway in a subset of lung cancer cell lines that are initially sensitive to EGFR-specific TKIs. The findings support FGFR-specific TKIs as potentially valuable additions to existing targeted therapeutic strategies with EGFR-specific TKIs to prevent or delay acquired resistance in EGFR-driven NSCLC. Examination of mRNA levels in DMSO and gefitinib-resistant cultures of HCC4006 and HCC827. Each group has two replicates.