BACKGROUND: RNA polymerase I (Pol I) transcription of ribosomal RNA genes within the nucleoli is consistently upregulated in cancer. Our laboratory developed CX-5461, a highly selective inhibitor of Pol I transcription [1]. Our preclinical studies demonstrated that single-agent treatment with CX-5461 provides a significant survival benefit in various pre-clinical cancer models [1-3]. Our first-in-human study has also shown a promising anti-tumour activity in three of six heavily pre-treated multiple myeloma (MM) patients, supporting CX-5461 as a promising treatment for MM [4]. However, drug resistance eventually occurs, indicating combination drug therapy is essential.
AIM: To examine the efficacy of CX-5461 as a single agent and in combination with other agents having proven clinical or promising preclinical efficacy in MM.
METHODS: We measured the effects of CX-5461 in human myeloma cell lines (HMCLs), 5T33 murine myeloma cells and MM patient samples. CX-5461 efficacy was tested in NSG and C57BL/KaLwRij mice transplanted with 5T33 cells. We conducted a high-throughput screen in HMCLs of CX-5461 combined with different agents. The effect of combining CX-5461 with the histone deacetylase inhibitor (HDACi) panobinostat was tested in vivo using Vk*MYC and 5T33-KaLwRij MM murine models.
RESULTS: CX-5461 showed anti-proliferative effects and cell death in vitro, with proteomic studies exhibiting enrichment of factors involved in cell cycle control and DNA damage response. There was increased survival benefit in immunocompetent 5T33-KaLwRij model treated with CX-5461, but not in immunodeficient NSG mice. Peripheral blood mononuclear cells from MM patient treated with CX-5461 displayed increased levels of γH2AX and cleaved caspase 3. Panobinostat demonstrated the most impressive synergy with CX-5461. Using both the MYC-driven Vk*MYC model and the non-MYC-driven 5T33-KaLwRij models of MM, we showed that CX-5461 with panobinostat provides a significant survival advantage.
CONCLUSION: CX-5461 enhances cell cycle arrest, DNA damage and cell death and exhibits immuno-stimulatory activities in pre-clinical MM models. Combining CX-5461 with panobinostat produces a survival advantage in both the Vk*MYC and the 5T33-KaLwRij models of MM. These results will direct subsequent clinical trials using CX-5461 in combination drug therapy.