Afatinib restrains K-RAS–driven lung tumorigenesis

Abstract

On the basis of clinical trials using first-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs), it became a doctrine that V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog () mutations drive resistance to EGFR inhibition in non-small cell lung cancer (NSCLC). Conversely, we provide evidence that EGFR signaling is engaged in K-RAS-driven lung tumorigenesis in humans and in mice. Specifically, genetic mouse models revealed that deletion of quenches mutant K-RAS activity and transiently reduces tumor growth. However, EGFR inhibition initiates a rapid resistance mechanism involving non-EGFR ERBB family members. This tumor escape mechanism clarifies the disappointing outcome of first-generation TKIs and suggests high therapeutic potential of pan-ERBB inhibitors. On the basis of various experimental models including genetically engineered mouse models, patient-derived and cell line-derived xenografts, and in vitro experiments, we demonstrate that the U.S. Food and Drug Administration-approved pan-ERBB inhibitor afatinib effectively impairs K-RAS-driven lung tumorigenesis. Our data support reconsidering the use of pan-ERBB inhibition in clinical trials to treat-mutated NSCLC.

JAK–STAT inhibition impairs K‐RAS‐driven lung adenocarcinoma progression

“…Intriguingly, global gene expression analysis revealed that K‐ras G12D expression resulted in activation of the JAK–STAT pathway (Figs. S3 a and S3 b ) . Encouraged by these data, we treated K and KP mice 1 week after tumor induction with ruxolitinib or vehicle control for a period of 10 weeks, euthanized them and analyzed their lungs.…”

“…For subcutaneous xenograft experiments using the K‐RAS ‐mutated human lung AC cell lines A549 and A427, 1 × 10 6 cells were mixed 1:2 with Matrigel (Corning, Corning, NY) and injected into the right and left flank of immunodeficient NOD scid gamma mice older than 6 weeks of age. Tumor volume was measured using a caliper and tumor volume was calculated using the formula [(length × width 2 ) × 0.52] as previously established in our laboratory …”

“…Data were analyzed using the web‐based online tool InteractiVenn, and for hierarchical clustering and heatmap illustration, we used the heatmapper.ca web tool . In order to check for the activation status of the JAK–STAT pathway and the expression of chemo/cytokine related genes in primary K‐ras G12D ‐mutated lung alveolar type II (K) cells, we analyzed the publically available GSE113146 data set …”

“…Tumor volume was measured using a caliper and tumor volume was calculated using the formula [(length × width 2 ) × 0.52] as previously established in our laboratory. 27 K-ras LSL-G12D (K) mice as well as K-ras LSL-G12D :p53 fl/fl (KP) mice, both bred on a C57Bl/6 background, served as genetically engineered mouse models for autochthonous lung AC. 28 Genotyping of the transgenes was performed using primers depicted in Table S3.…”

“…33 In order to check for the activation status of the JAK-STAT pathway and the expression of chemo/cytokine related genes in primary K-ras G12D -mutated lung alveolar type II (K) cells, we analyzed the publically available GSE113146 data set. 27…”