Data from Ruthenium Drug BOLD-100 Regulates <i>BRAF</i>MT Colorectal Cancer Cell Apoptosis through AhR/ROS/ATR Signaling Axis Modulation

Abstract

<div>Abstract<p>Patients with class I <sup>V600E</sup><i>BRAF</i>-mutant (MT) colorectal cancer exhibit a poor prognosis, and their response to combined anti-BRAF/EGFR inhibition remains limited. An unmet need exits for further understanding the biology of <sup>V600E</sup><i>BRAF</i>MT colorectal cancer. We used differential gene expression of <i>BRAF</i>WT and MT colorectal cancer cells to identify pathways underpinning <i>BRAF</i>MT colorectal cancer. We tested a panel of molecularly/genetically subtyped colorectal cancer cells for their sensitivity to the unfolded protein response (UPR) activator BOLD-100. To identify novel combination strategies for BOLD-100, we performed RNA sequencing and high-throughput drug screening. Pathway enrichment analysis identified significant enrichment of the UPR and DNA repair pathways in <i>BRAF</i>MT colorectal cancer. We found that oncogenic <i>BRAF</i> plays a crucial role in mediating the response to BOLD-100. Using a systems biology approach, we identified <sup>V600E</sup><i>BRAF</i>MT-dependent activation of the replication stress response kinase ataxia telangiectasia and Rad3-related (ATR) as a key mediator of resistance to BOLD-100. Further analysis identified acute increases in <i>BRAF</i>MT-dependent-reactive oxygen species levels following treatment with BOLD-100, which promoted ATR/CHK1 activation and apoptosis. Furthermore, activation of reactive oxygen species/ATR/CHK1 following BOLD-100 was mediated through the AhR transcription factor and CYP1A1. Importantly, pharmacological blockade of this resistance pathway with ATR inhibitors synergistically increased BOLD-100-induced apoptosis and growth inhibition in <i>BRAF</i>MT models. These results highlight a possible novel therapeutic opportunity for <i>BRAF</i>MT colorectal cancer.</p><p><b>Implications:</b> BOLD-100 induces <i>BRAF</i>MT-dependent replication stress, and targeted strategies against replication stress (e.g., by using ATR inhibitors) in combination with BOLD-100 may serve as a potential novel therapeutic strategy for clinically aggressive <i>BRAF</i>MT colorectal cancer.</p></div>