scholarly journals YES1amplification: a mechanism of acquired resistance to EGFR inhibitors identified by transposon mutagenesis and clinical genomics

2018 ◽  
Author(s):  
Pang-Dian Fan ◽  
Giuseppe Narzisi ◽  
Anitha D. Jayaprakash ◽  
Elisa Venturini ◽  
Nicolas Robine ◽  
...  
Keyword(s):  
Cell Line ◽  
Mutant Cell ◽  
Acquired Resistance ◽  
Transposon Mutagenesis ◽  
Egfr Inhibitors ◽  
Genomic Sequencing ◽  
Mechanisms Of Resistance ◽  
Lung Cancers ◽  
Lung Adenocarcinomas ◽  
Clinical Genomic

ABSTRACTIn approximately 30% of patients withEGFR-mutant lung adenocarcinomas whose disease progresses on EGFR inhibitors, the basis for acquired resistance remains unclear. We have integrated transposon mutagenesis screening in anEGFR-mutant cell line and clinical genomic sequencing in cases of acquired resistance to identify novel mechanisms of resistance to EGFR inhibitors. The most prominent candidate genes identified by insertions in or near the genes during the screen wereMET, a gene whose amplification is known to mediate resistance to EGFR inhibitors, and the gene encoding the Src family kinase YES1. Cell clones with transposon insertions that activated expression ofYES1exhibited resistance to all three generations of EGFR inhibitors and sensitivity to pharmacologic and siRNA-mediated inhibition ofYES1. Analysis of clinical genomic sequencing data from cases of acquired resistance to EGFR inhibitors revealed amplification ofYES1in 5 cases, 4 of which lacked any other known mechanisms of resistance. Pre-inhibitor samples, available for 2 of the 5 patients, lackedYES1amplification. None of 136 post-inhibitor samples had detectable amplification of other Src family kinases (SRC, FYN).YES1amplification was also found in 2 of 17 samples fromALKfusion-positive lung cancer patients who had progressed on ALK TKIs. Taken together, our findings identify acquired amplification ofYES1as a novel, recurrent, and targetable mechanism of resistance to EGFR inhibition inEGFR-mutant lung cancers, and demonstrate the utility of transposon mutagenesis in discovering clinically relevant mechanisms of drug resistance.SIGNIFICANCEDespite high response rates to treatment with small molecule inhibitors of EGFR tyrosine kinase activity, patients withEGFR-mutant lung adenocarcinomas eventually develop resistance to these drugs. In many cases, the basis of acquired resistance remains unclear. We have used a transposon mutagenesis screen in anEGFR-mutant cell line and clinical genomic sequencing in cases of acquired resistance to identify amplification ofYES1as a novel and targetable mechanism of resistance to EGFR inhibitors inEGFR-mutant lung cancers.

scholarly journals YES1amplification is a mechanism of acquired resistance to EGFR inhibitors identified by transposon mutagenesis and clinical genomics

2018 ◽  
Vol 115 (26) ◽  
pp. E6030-E6038 ◽  
Author(s):  
Pang-Dian Fan ◽  
Giuseppe Narzisi ◽  
Anitha D. Jayaprakash ◽  
Elisa Venturini ◽  
Nicolas Robine ◽  
...  
Keyword(s):  
Mutant Cell ◽  
Acquired Resistance ◽  
Transposon Mutagenesis ◽  
Egfr Inhibitors ◽  
Genomic Sequencing ◽  
Sequencing Data ◽  
Mechanisms Of Resistance ◽  
Lung Cancers ◽  
Egfr Inhibition ◽  
Clinical Genomic

In ∼30% of patients withEGFR-mutant lung adenocarcinomas whose disease progresses on EGFR inhibitors, the basis for acquired resistance remains unclear. We have integrated transposon mutagenesis screening in anEGFR-mutant cell line and clinical genomic sequencing in cases of acquired resistance to identify mechanisms of resistance to EGFR inhibitors. The most prominent candidate genes identified by insertions in or near the genes during the screen wereMET, a gene whose amplification is known to mediate resistance to EGFR inhibitors, and the gene encoding the Src family kinase YES1. Cell clones with transposon insertions that activated expression ofYES1exhibited resistance to all three generations of EGFR inhibitors and sensitivity to pharmacologic and siRNA-mediated inhibition ofYES1. Analysis of clinical genomic sequencing data from cases of acquired resistance to EGFR inhibitors revealed amplification ofYES1in five cases, four of which lacked any other known mechanisms of resistance. Preinhibitor samples, available for two of the five patients, lackedYES1amplification. None of 136 postinhibitor samples had detectable amplification of other Src family kinases (SRCandFYN).YES1amplification was also found in 2 of 17 samples fromALKfusion-positive lung cancer patients who had progressed on ALK TKIs. Taken together, our findings identify acquired amplification ofYES1as a recurrent and targetable mechanism of resistance to EGFR inhibition inEGFR-mutant lung cancers and demonstrate the utility of transposon mutagenesis in discovering clinically relevant mechanisms of drug resistance.

Abstract 955: Transformation from NSCLC to SCLC in EGFR mutant lung cancers with acquired resistance to EGFR inhibitors

2014 ◽  
Author(s):  
Matthew J. Niederst ◽  
Lecia V. Sequist ◽  
Elizabeth L. Lockerman ◽  
Angel R. Garcia ◽  
Carlotta Costa ◽  
...  
Keyword(s):  
Acquired Resistance ◽  
Egfr Inhibitors ◽  
Lung Cancers ◽  
Egfr Mutant

YES1 amplification as a mechanism of acquired resistance (AR) to EGFR tyrosine kinase inhibitors (TKIs) identified by a transposon mutagenesis screen and clinical genomic testing.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. 9043-9043 ◽  
Author(s):  
Pang-Dian Fan ◽  
Giuseppe Narzisi ◽  
Anitha Jayaprakash ◽  
Elisa Venturini ◽  
Nicolas Robine ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Kinase Inhibitors ◽  
Acquired Resistance ◽  
Transposon Mutagenesis ◽  
Src Family Kinase ◽  
Mechanisms Of Resistance ◽  
Transposon Insertion ◽  
Insertion Sites ◽  
Egfr Tki ◽  
Egfr Tkis

9043 Background: Overcoming AR to EGFR TKIs remains challenging, and in many cases the mechanisms are still unclear. To identify novel mechanisms of resistance to EGFR TKIs, we performed a forward genetic screen using transposon mutagenesis in EGFR-mutant lung adenocarcinoma cells. Methods: EGFR TKI-sensitive PC9 cells were co-transfected with plasmids encoding a mutagenic piggyBactransposon and hyperactive piggyBac transposase. Transposon-tagged, afatinib-resistant clones were generated by sequential selection of transfected cells with puromycin and 1µM afatinib. Transposon insertion sites were mapped using a modified TraDIS-type method and next-generation sequencing (NGS). Selected clones were characterized using Western blots, receptor tyrosine kinase (RTK) arrays, and viability assays following treatment with TKIs or siRNA-mediated gene knockdowns. We reviewed MSK-IMPACT™ NGS data on 100 patient tumors with EGFR TKI AR. Available tumor samples were analyzed by fluorescence in situ hybridization (FISH). Results: In 187/188 afatinib-resistant clones, transposon insertion sites consistent predominantly with gene upregulation were found in MET, the Src family kinase (SFK) member YES1, or both. Clones with activating YES1 insertions exhibited resistance to all three generations of EGFR TKIs; high levels of expression of tyrosine-phosphorylated YES1; sensitivity to the SFK TKI dasatinib and to siRNA-mediated knockdown of YES1; and tyrosine phosphorylation of YAP1 and ERBB3. A query of the MSK-IMPACT™ data on EGFR TKI AR patients revealed amplification of YES1 and no alteration of MET, ERBB2 or BRAF in 3/54 T790M-negative (95% CI 1 to 16%) and 1/46 (95% CI 1 to 12%) T790M-positive cases. Amplification of YES1was confirmed by FISH in 2/2 cases, and was absent in matched pre-TKI samples in 2/2 cases. Conclusions: YES1 amplification is found in 4% of patients with acquired resistance to EGFR TKIs and is potentially targetable by Src family kinase inhibitors. Forward genetic screens using transposon mutagenesis and routine clinical NGS of patient samples can identify novel mechanisms of resistance to targeted therapies.

scholarly journals Lung cancers with acquired resistance to EGFR inhibitors occasionally harbor BRAF gene mutations but lack mutations in KRAS, NRAS, or MEK1

2012 ◽  
Vol 109 (31) ◽  
pp. E2127-E2133 ◽  
Author(s):  
K. Ohashi ◽  
L. V. Sequist ◽  
M. E. Arcila ◽  
T. Moran ◽  
J. Chmielecki ◽  
...  
Keyword(s):  
Acquired Resistance ◽  
Gene Mutations ◽  
Egfr Inhibitors ◽  
Braf Gene ◽  
Lung Cancers

scholarly journals Cell Line Models for Acquired Resistance to First-Line Osimertinib in Lung Cancers—Applications and Limitations

Cells ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 354
Author(s):  
Shuta Ohara ◽  
Kenichi Suda ◽  
Tetsuya Mitsudomi
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Second Generation ◽  
Acquired Resistance ◽  
Research Groups ◽  
First Line ◽  
Lung Cancers ◽  
Mesenchymal Transition ◽  
First And Second Generation ◽  
Egfr Tkis

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are first-line drugs for lung cancers with activating EGFR mutations. Although first- and second-generation EGFR-TKIs were standard first-line treatments, acquired resistance (AR) to these drugs is almost inevitable. Cell line models have been widely used to explore the molecular mechanisms of AR to first- and second-generation EGFR-TKIs. Many research groups, including ours, have established AR cell lines that harbor the EGFR T790M secondary mutation, MET gene amplification, or epithelial–mesenchymal transition (EMT) features, which are all found in clinical specimens obtained from TKI-refractory lesions. Currently, many oncologists prescribe osimertinib, a third-generation EGFR-TKI that can overcome T790M-mediated resistance, as a first-line TKI. Although few clinical data are available about AR mechanisms that arise when osimertinib is used as a first-line therapy, many research groups have established cell lines with AR to osimertinib and have reported on their AR mechanisms. In this review, we summarize the findings on AR mechanisms against first-line osimertinib obtained from analyses of cell line models.

Tissue-based molecular and histological landscape of acquired resistance to osimertinib given initially or at relapse in patients with EGFR-mutant lung cancers.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. 9028-9028 ◽  
Author(s):  
Adam Jacob Schoenfeld ◽  
Joseph Minhow Chan ◽  
Hira Rizvi ◽  
Natasha Rekhtman ◽  
Yahya Daneshbod ◽  
...  
Keyword(s):  
Initial Treatment ◽  
Tumor Tissue ◽  
Cell Transformation ◽  
Chromosomal Rearrangements ◽  
Acquired Resistance ◽  
Resistance Mechanisms ◽  
Resistance Mutations ◽  
Mechanisms Of Resistance ◽  
Lung Cancers ◽  
Egfr Mutant

9028 Background: Even though osimertinib (osi) is now the initial treatment for patients with EGFR-mutant lung cancers, our knowledge about mechanisms of resistance (MOR) is largely derived from patients who received osi after developing acquired resistance to initial EGFR inhibitor. Further, studies of osi resistance to date have mainly reported genotyping of plasma which suboptimally detects lineage plasticity, copy number changes, and chromosomal rearrangements. Methods: To identify MOR to osi and characterize clinical, molecular and histologic factors associated with duration of response, we identified patients with EGFR-mutant lung cancers who had next-generation sequencing performed on tumor tissue after developing acquired resistance to osi. Results: From January 2016 to December 2018, post-osi tumor tissue was collected from 71 patients (42 with paired pre-treatment specimens). See mechanisms of resistance below. Histologic transformation was identified in 19% of initial cases and 14% of all cases. When osi is given as initial treatment, with median follow up of 17 months, early emerging MOR rarely included on-target resistance mechanisms (1/16 cases of acquired EGFR G724S). Acquired alterations representing potential resistance mechanisms included CCNE1 and MYC amplifications, and mutations in MTOR and MET H1094Y. We confirmed in preclinical studies that an amino acid substitution at MET H1094 can reduce sensitivity to osi. Conclusions: In this analysis of MOR identified on NGS from tumor tissue, we found a different spectrum of resistance mechanisms to initial and later-line osi, with histologic transformation (including squamous cell transformation) a dominant MOR, particularly in the first-line setting, that cannot be identified on plasma testing. Subsequent studies are needed to assess patients with a longer time on initial osi as there may be a temporal bias to MOR, with off-target MOR emerging earlier and on-target resistance mutations later. [Table: see text]

Managing Resistance to EFGR- and ALK-Targeted Therapies

2017 ◽  
pp. 607-618
Author(s):  
Christine M. Lovly ◽  
Puneeth Iyengar ◽  
Justin F. Gainor
Keyword(s):  
Targeted Therapies ◽  
Molecular Mechanisms ◽  
Acquired Resistance ◽  
Growth Factor Receptor ◽  
Genetic Alterations ◽  
Mechanisms Of Resistance ◽  
Lung Cancers ◽  
Anaplastic Lymphoma ◽  
Ablative Therapies ◽  
Egfr Mutant

Targeted therapies have transformed the management of non–small cell lung cancer (NSCLC) and placed an increased emphasis on stratifying patients on the basis of genetic alterations in oncogenic drivers. To date, the best characterized molecular targets in NSCLC are the epidermal growth factor receptor (EGFR) and anaplastic lymphoma kinase (ALK). Despite steady advances in targeted therapies within these molecular subsets, however, acquired resistance to therapy is near universal. Recent preclinical models and translational efforts have provided critical insights into the molecular mechanisms of resistance to EGFR and ALK inhibitors. In this review, we present a framework for understanding resistance to targeted therapies. We also provide overviews of the molecular mechanisms of resistance and strategies to overcome resistance among EGFR-mutant and ALK-rearranged lung cancers. To date, these strategies have centered on the development of novel next-generation inhibitors, rationale combinations, and use of local ablative therapies, such as radiotherapy.

The tyrosine kinase TEK is differentially expressed in non-small cell lung adenocarcinomas.

2020 ◽  
Author(s):  
Shahan Mamoor
Keyword(s):  
United States ◽  
Tyrosine Kinase ◽  
Lung Adenocarcinoma ◽  
Microarray Data ◽  
The United States ◽  
Small Cell ◽  
Small Cell Lung ◽  
Significant Differential Expression ◽  
Lung Cancers ◽  
Lung Adenocarcinomas

Non-small cell lung adenocarcinoma (NSCLC) is a leading cause of death in the United States and worldwide (1, 2). We mined published microarray data (3, 4, 5) to discover genes associated with NSCLC. We identified significant differential expression of the tyrosine kinase TEK in tumors from patients with NSCLC. TEK may be of relevance to the initiation, progression or maintenance of non-small cell lung cancers.

Sign in / Sign up

Export Citation Format

Share Document