MIG6 loss confers resistance to ALK/ROS1 inhibitors in NSCLC through EGFR activation by low-dose EGF
Nobuyuki Kondo, Takahiro Utsumi, Yuki Shimizu, Ai Takemoto, Tomoko Oh-hara, Ken Uchibori, Sophia Subat-Motoshi, Hironori Ninomiya, Kengo Takeuchi, Makoto Nishio, Yasunari Miyazaki, Ryohei Katayama
Nobuyuki Kondo, Takahiro Utsumi, Yuki Shimizu, Ai Takemoto, Tomoko Oh-hara, Ken Uchibori, Sophia Subat-Motoshi, Hironori Ninomiya, Kengo Takeuchi, Makoto Nishio, Yasunari Miyazaki, Ryohei Katayama
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Research Article Oncology

MIG6 loss confers resistance to ALK/ROS1 inhibitors in NSCLC through EGFR activation by low-dose EGF

Abstract

Although tyrosine kinase inhibitor (TKI) therapy shows marked clinical efficacy in patients with anaplastic lymphoma kinase–positive (ALK+) and ROS proto-oncogene 1–positive (ROS1+) non–small cell lung cancer (NSCLC), most of these patients eventually relapse with acquired resistance. Therefore, genome-wide CRISPR/Cas9 knockout screening was performed using an ALK+ NSCLC cell line established from pleural effusion without ALK-TKI treatment. After 9 days of ALK-TKI therapy, sequencing analysis was performed, which identified several tumor suppressor genes, such as NF2 or MED12, and multiple candidate genes. Among them, this study focused on ERRFI1, which is known as MIG6 and negatively regulates EGFR signaling. Interestingly, MIG6 loss induced resistance to ALK-TKIs by treatment with quite a low dose of EGF, which is equivalent to plasma concentration, through the upregulation of MAPK and PI3K/AKT/mTOR pathways. Combination therapy with ALK-TKIs and anti-EGFR antibodies could overcome the acquired resistance in both in vivo and in vitro models. In addition, this verified that MIG6 loss induces resistance to ROS1-TKIs in ROS1+ cell lines. This study found a potentially novel factor that plays a role in ALK and ROS1-TKI resistance by activating the EGFR pathway with low-dose ligands.

Authors

Nobuyuki Kondo, Takahiro Utsumi, Yuki Shimizu, Ai Takemoto, Tomoko Oh-hara, Ken Uchibori, Sophia Subat-Motoshi, Hironori Ninomiya, Kengo Takeuchi, Makoto Nishio, Yasunari Miyazaki, Ryohei Katayama

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Figure 3

Combination therapy with EGFR inhibitors and ALK-TKIs can overcome the resistance related to MIG6 depletion.

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Combination therapy with EGFR inhibitors and ALK-TKIs can overcome the r...
(A) Colony formation assays were performed in JFCR-028-3 cells. Each well was treated with 100 nmol/L of alectinib or 30 nmol/L of lorlatinib with or without 10 μg/mL of panitumumab for 2 weeks, and surviving cells were stained with crystal violet. Representative images are shown. (B and C) JFCR-028-3 cells were treated with the indicated concentrations of ALK-TKIs and ligands with or without 10 μg/mL of panitumumab (B) or 100 nmol/L of afatinib (C) for 72 hours. Cell viability was measured using the CellTiter-Glo assay. Each point represents the mean ± SD of 3 technical replicates. (D) Protein expression of the downstream pathway of ALK in JFCR-028-3 cells. Cells were treated with 300 nmol/L of alectinib, 10 μg/mL of panitumumab, 100 nmol/L of afatinib, and 1 ng/mL of EGF for 3 hours. (A–D) Similar experiments were performed twice (D) or 3 times (A–C), and representative data are shown.