TY - JOUR
T1 - Resistance Profile and Structural Modeling of Next-Generation ROS1 Tyrosine Kinase Inhibitors
AU - Keddy, Clare
AU - Shinde, Pushkar
AU - Jones, Kristen
AU - Kaech, Stefanie
AU - Somwar, Romel
AU - Shinde, Ujwal
AU - Davare, Monika A.
N1 - Funding Information:
R. Somwar reports grants from Helsinn Health Care, Loxo Oncology, Merus, Elevation Oncology, and non-financial support from GlaxoSmithKline outside the submitted work. M.A. Davare reports grants from Nuvalent outside the submitted work, as well as a patent for WO2014134096A1 issued and a patent 10100366 issued. No disclosures were reported by the other authors.
Funding Information:
We would like to thank Dr. Nicolle Hofmann for assistance with cloning the mYFP tagged CD74-, EZR-, SLC34A2-, and GOPC-ROS1 constructs, and Sudarshan Iyer for assisting with generation of pENTR-EZR-ROS1 construct. This work was partially funded by R01 CA233495-01A1 and an American Cancer Society (ACS) grant (RSG-19-082-01-TBG) to M.A. Davare.
Publisher Copyright:
© 2021 American Association for Cancer Research.
PY - 2022/2
Y1 - 2022/2
N2 - ROS1 fusion proteins resulting from chromosomal rearrangements of the ROS1 gene are targetable oncogenic drivers in diverse cancers. Acquired resistance to targeted inhibitors curtails clinical benefit and response durability. Entrectinib, a NTRK/ROS1/ALK targeted tyrosine kinase inhibitor (TKI), was approved for the treatment of ROS1 fusion-positive non–small cell lung cancer (NSCLC) in 2019. In addition, lorlatinib and repotrectinib are actively being explored in the setting of treatment-naïve or crizotinib-resistant ROS1 fusion driven NSCLC. Here, we employed an unbiased forward mutagenesis screen in Ba/F3 CD74-ROS1 and EZR-ROS1 cells to identify resistance liabilities to entrectinib, lorlatinib, and repotrectinib. ROS1F2004C emerged as a recurrent entrectinib resistant mutation and ROS1G2032R was discovered in entrectinib and lorlatinib-resistant clones. Cell-based and modeling data show that entrectinib is a dual type I/II mode inhibitor, and thus liable to both types of resistant mutations. Comprehensive profiling of all clinically relevant kinase domain mutations showed that ROS1L2086F is broadly resistant to all type I inhibitors, but remains sensitive to type II inhibitors. ROS1F2004C/I/V are resistant to type I inhibitors, entrectinib and crizotinib, and type II inhibitor, cabozantinib, but retain sensitivity to the type I macrocyclic inhibitors. Development of new, more selective type II ROS1 inhibitor (s) or potentially cycling type I and type II inhibitors may be one way to expand durability of ROS1-targeted agents.
AB - ROS1 fusion proteins resulting from chromosomal rearrangements of the ROS1 gene are targetable oncogenic drivers in diverse cancers. Acquired resistance to targeted inhibitors curtails clinical benefit and response durability. Entrectinib, a NTRK/ROS1/ALK targeted tyrosine kinase inhibitor (TKI), was approved for the treatment of ROS1 fusion-positive non–small cell lung cancer (NSCLC) in 2019. In addition, lorlatinib and repotrectinib are actively being explored in the setting of treatment-naïve or crizotinib-resistant ROS1 fusion driven NSCLC. Here, we employed an unbiased forward mutagenesis screen in Ba/F3 CD74-ROS1 and EZR-ROS1 cells to identify resistance liabilities to entrectinib, lorlatinib, and repotrectinib. ROS1F2004C emerged as a recurrent entrectinib resistant mutation and ROS1G2032R was discovered in entrectinib and lorlatinib-resistant clones. Cell-based and modeling data show that entrectinib is a dual type I/II mode inhibitor, and thus liable to both types of resistant mutations. Comprehensive profiling of all clinically relevant kinase domain mutations showed that ROS1L2086F is broadly resistant to all type I inhibitors, but remains sensitive to type II inhibitors. ROS1F2004C/I/V are resistant to type I inhibitors, entrectinib and crizotinib, and type II inhibitor, cabozantinib, but retain sensitivity to the type I macrocyclic inhibitors. Development of new, more selective type II ROS1 inhibitor (s) or potentially cycling type I and type II inhibitors may be one way to expand durability of ROS1-targeted agents.
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U2 - 10.1158/1535-7163.MCT-21-0395
DO - 10.1158/1535-7163.MCT-21-0395
M3 - Article
C2 - 34907086
AN - SCOPUS:85124438270
VL - 21
SP - 336
EP - 346
JO - Molecular Cancer Therapeutics
JF - Molecular Cancer Therapeutics
SN - 1535-7163
IS - 2
ER -