TERRA G-quadruplex stabilization as a new therapeutic strategy for multiple myeloma

Abstract

AbstractBackgroundMultiple myeloma (MM) is a hematologic malignancy characterized by high genomic instability, and telomere dysfunction is an important cause of acquired genomic alterations. Telomeric repeat-containing RNA (TERRA) transcripts are long non-coding RNAs involved in telomere stability through the interaction with shelterin complex. Dysregulation of TERRAs has been reported across several cancer types. We recently identified a small molecule,hit17, which stabilizes the secondary structure of TERRA. In this study, we investigated in vitro and in vivo anti-MM activities ofhit17.MethodsAnti-proliferative activity ofhit17 was evaluated in different MM cell lines by cell proliferation assay, and the apoptotic process was analyzed by flow cytometry. Gene and protein expressions were detected by RT-qPCR and western blotting, respectively. Microarray analysis was used to analyze the transcriptome profile. The effect ofhit17 on telomeric structure was evaluated by chromatin immunoprecipitation. Further evaluation in vivo was proceeded upon NCI-H929 and AMO-1 xenograft models.ResultsTERRA G4 stabilization induced in vitro dissociation of telomeric repeat‐binding factor 2 (TRF2) from telomeres leading to the activation of ATM-dependent DNA damage response, cell cycle arrest, proliferation block, and apoptotic death in MM cell lines. In addition, up-regulation of TERRA transcription was observed upon DNA damage and TRF2 loss. Transcriptome analysis followed by gene set enrichment analysis (GSEA) confirmed the involvement of the above-mentioned processes and other pathways such as E2F, MYC, oxidative phosphorylation, and DNA repair genes as early events followinghit17-induced TERRA stabilization. Moreover,hit17 exerted anti-tumor activity against MM xenograft models.ConclusionOur findings provide evidence that targeting TERRA byhit17 could represent a promising strategy for a novel therapeutic approach to MM.