Abstract P2-05-05: RAD51 inhibition using CYT-0851, shows anti-cancer activity in cellular models of breast cancer and acts synergistically with PARP inhibitors
Authors: Joey L Guy, Tyler Maclay, Melinda Day, Monika L Burness, Kevin Mills
Journal: Cancer Research (2020)

Abstract Genomic instability is recognized as a driver of tumorigenesis and cancer progression. Loss of tumor suppressors or activation of oncogenes can induce DNA damage stress, promoting genomic instability and creating dependencies upon key DNA repair pathways. The clinical success of PARP inhibitors has highlighted the potential of targeting these dependencies therapeutically to induce synthetic lethality. We have developed novel RAD51 inhibitors to selectively target cancers that have elevated levels of DNA damage and/or reduced levels of DNA repair. RAD51 is a critical component of the homologous recombination (HR) pathway, forming nucleoprotein filaments at sites of DNA damage or replication fork stalls. RAD51 mediates homologous DNA strand exchange to promote recombinational repair of breaks and damaged replication forks. We have previously shown anti-cancer activity of RAD51 inhibitors in preclinical xenograft mouse models of lymphomas that have elevated rates of DNA mutations due to aberrant expression of Activation Induced Cytidine Deaminase (AID). Breast cancer, due to high rates of genomic instability, may potentially be treated in this fashion. Here we present the characterization of our RAD51 inhibitors in breast cancer. In a murine cell line model of metastatic breast cancer, 4T1, treatment with CYT-0851 resulted in a reduction in homologous recombination activity in a sister chromatid exchange (SCE) assay at a similar rate observed in a human derived cell line (HEK293T) suggesting conservation of mechanism in human and murine models. In the same cell line, treatment with CYT-0851 and a DNA damaging agent, carboplatin, resulted in cellular toxicity above what was observed with CYT-0851, indicating that RAD51 inhibition coupled with genomic instability is cytotoxic in this breast cancer model. PARP inhibitors have been shown to induce genomic instability as measured by an increase in HR activity in SCE assays. We therefore hypothesized that our RAD51 inhibitor could act as a sensitizer to PARP inhibitors. Combinatorial treatment of 4T1 cells with CYT-0851 and olaparib reduced homologous recombination activity in the SCE assay. Additionally, combination therapy led to increased accumulation of gH2AX foci after ionizing radiation, and increased tail moment on neutral Comet assay. Chou-Talalay synergy assay demonstrated synergistic activity between olaparib and CYT-0851 in 4T1. To determine potential drug synergy in human models, a matrix study was performed with CYT-0851 and olaparib in three cell lines of triple negative breast cancer (HCC1937, HCC1143, and BT20). Concentration-dependent synergy was observed in all cell lines tested. These data suggest that CYT-0851 may be active as a mono therapy and a combinatorial therapy with PARP inhibitors in breast cancer. Overall, we conclude that the synergy observed in vitro indicates significant potential for RAD51 inhibition in future cancer treatment strategies and warrants future exploration in vivo. Citation Format: Joey L Guy, Tyler Maclay, Melinda Day, Monika L Burness, Kevin Mills. RAD51 inhibition using CYT-0851, shows anti-cancer activity in cellular models of breast cancer and acts synergistically with PARP inhibitors [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P2-05-05.