Synthetic Lethal Screen Identifies NF-kappaB as a Target for Combination Therapy with Topotecan for patients with Neuroblastoma
BACKGROUND:Despite aggressive multimodal treatments the overall survival of patients with high-risk neuroblastoma remains poor. The aim of this study was to identify novel combination chemotherapy to improve survival rate in patients with high-risk neuroblastoma.METHODS:We took a synthetic lethal approach using a siRNA library targeting 418 apoptosis-related genes and identified genes and pathways whose inhibition synergized with topotecan. Microarray analyses of cells treated with topotecan were performed to identify if the same genes or pathways were altered by the drug. An inhibitor of this pathway was used in combination with topotecan to confirm synergism by in vitro and in vivo studies.RESULTS:We found that there were nine genes whose suppression synergized with topotecan to enhance cell death, and the NF-kappaB signaling pathway was significantly enriched. Microarray analysis of cells treated with topotecan revealed a significant enrichment of NF-kappaB target genes among the differentially altered genes, suggesting that NF-kappaB pathway was activated in the treated cells. Combination of topotecan and known NF-kappaB inhibitors (NSC 676914 or bortezomib) significantly reduced cell growth and induced caspase 3 activity in vitro. Furthermore, in a neuroblastoma xenograft mouse model, combined treatment of topotecan and bortezomib significantly delayed tumor formation compared to single-drug treatments.CONCLUSIONS:Synthetic lethal screening provides a rational approach for selecting drugs for use in combination therapy and warrants clinical evaluation of the efficacy of the combination of topotecan and bortezomib or other NF-kappaB inhibitors in patients with high risk neuroblastoma.