Discovery of oncogenic ROS1 missense mutations with sensitivity to tyrosine kinase inhibitors

Abstract

ROS1 is the largest receptor tyrosine kinase in the human genome. Rearrangements of the ROS1 gene result in oncogenic ROS1 kinase fusion proteins that are currently the only validated biomarkers for targeted therapy with ROS1 TKIs in patients. While numerous somatic missense mutations in ROS1 exist in the cancer genome, their impact on catalytic activity and pathogenic potential is unknown. We interrogated the AACR Genie database and identified 34 missense mutations in the ROS1 tyrosine kinase domain for further analysis. Our experiments revealed that these mutations have varying effects on ROS1 kinase function, ranging from complete loss to significantly increased catalytic activity. Notably, Asn and Gly substitutions at Asp2113 in the ROS1 kinase domain were found to be TKI‐sensitive oncogenic variants in cell‐based model systems. In vivo experiments showed that ROS1 D2113N induced tumor formation that was sensitive to crizotinib and lorlatinib, FDA‐approved ROS1‐TKIs. Collectively, these findings highlight the tumorigenic potential of specific point mutations within the ROS1 kinase domain and their potential as therapeutic targets with FDA‐approved ROS1‐TKIs.

Kinase Inhibitors and Kinase-Targeted Cancer Therapies: Recent Advances and Future Perspectives

“…ROS1, a member of the RTK family, is a known oncogenic driver in various cancers, particularly lung adenocarcinoma. Despite being observed in only 1% to 2% of NSCLC patients [117], ROS1 plays a significant role in cancer through fusion proteins resulting from ROS1 rearrangements. These fusions, such as CD74-ROS1, SLC34A2-ROS1, and others, cause an uncontrolled activation of downstream pathways due to the loss of regulatory domains [118].…”

“…For instance, Iyer et al leveraged public databases to screen ROS1 variants, employing the "Sorting Intolerant From Tolerant" algorithm to assess the impact of mutations on protein function, particularly focusing on those with deleterious effects. They further evaluated the influence of mutations on ROS1 inhibitors through in vitro experiments [117]. Similarly, Zhao et al investigated the variations in binding affinity between the ROS1 protein and specific mutations like L2010M, G1957A, D1988N, and L1982V using molecular docking and molecular dynamics simulation techniques.…”

CRISPR/Cas9-edited ROS1 + non-small cell lung cancer cell lines highlight differential drug sensitivity in 2D vs 3D cultures while reflecting established resistance profiles

“…In contrast, HCC78 was the first available patient-derived ROS1 + cell line. It has been used in several studies and it is genetically well characterized [ 20 – 23 ]. Importantly, HCC78 spheroids have been used to assess the response to TKIs in ROS1 wild type cells [ 24 ].…”