Vlummens Philip, Verhulst Stefaan, De Veirman Kim, Maes Anke, Menu Eline, Moreaux Jérome, De Boussac Hugues, Robert Nicolas, De Bruyne Elke, Hose Dirk, Offner Fritz, Vanderkerken Karin, Maes Ken
Department of Hematology and Immunology, Vrije Universiteit Brussel (VUB), Brussels, Belgium.
Department of Clinical Hematology, Ghent University Hospital, Ghent, Belgium.
Front Cell Dev Biol. 2022 Jun 8;10:879057. doi: 10.3389/fcell.2022.879057. eCollection 2022.
Multiple myeloma (MM) is an incurable clonal plasma cell malignancy. Subsets of patients have high-risk features linked with dismal outcome. Therefore, the need for effective therapeutic options remains high. Here, we used bio-informatic tools to identify novel targets involved in DNA repair and epigenetics and which are associated with high-risk myeloma. The prognostic significance of the target genes was analyzed using publicly available gene expression data of MM patients (TT2/3 and HM cohorts). Hence, protein arginine methyltransferase 5 (PRMT5) was identified as a promising target. Druggability was assessed in OPM2, JJN3, AMO1 and XG7 human myeloma cell lines using the PRMT5-inhibitor EPZ015938. EPZ015938 strongly reduced the total symmetric-dimethyl arginine levels in all cell lines and lead to decreased cellular growth, supported by cell line dependent changes in cell cycle distribution. At later time points, apoptosis occurred, as evidenced by increased AnnexinV-positivity and cleavage of PARP and caspases. Transcriptome analysis revealed a role for PRMT5 in regulating alternative splicing, nonsense-mediated decay, DNA repair and PI3K/mTOR-signaling, irrespective of the cell line type. PRMT5 inhibition reduced the expression of upstream DNA repair kinases ATM and ATR, which may in part explain our observation that EPZ015938 and the DNA-alkylating agent, melphalan, have combinatory effects. Of interest, using a low-dose of mTOR-inhibitor, we observed that cell viability was partially rescued from the effects of EPZ015938, indicating a role for mTOR-related pathways in the anti-myeloma activity of EPZ015938. Moreover, PRMT5 was shown to be involved in splicing regulation of MMSET and SLAMF7, known genes of importance in MM disease. As such, we broaden the understanding of the exact role of PRMT5 in MM disease and further underline its use as a possible therapeutic target.
多发性骨髓瘤(MM)是一种无法治愈的克隆性浆细胞恶性肿瘤。部分患者具有与不良预后相关的高危特征。因此,对有效治疗方案的需求仍然很高。在此,我们使用生物信息学工具来识别参与DNA修复和表观遗传学且与高危骨髓瘤相关的新靶点。利用公开可得的MM患者基因表达数据(TT2/3和HM队列)分析了靶基因的预后意义。因此,蛋白精氨酸甲基转移酶5(PRMT5)被确定为一个有前景的靶点。使用PRMT5抑制剂EPZ015938在OPM2、JJN3、AMO1和XG7人骨髓瘤细胞系中评估了药物可及性。EPZ015938显著降低了所有细胞系中的总对称二甲基精氨酸水平,并导致细胞生长减少,细胞周期分布的细胞系依赖性变化对此有支持作用。在后期时间点,出现了凋亡,AnnexinV阳性增加以及PARP和半胱天冬酶的裂解证明了这一点。转录组分析揭示了PRMT5在调节可变剪接、无义介导的衰变、DNA修复和PI3K/mTOR信号传导中的作用,与细胞系类型无关。PRMT5抑制降低了上游DNA修复激酶ATM和ATR的表达,这可能部分解释了我们观察到的EPZ015938与DNA烷化剂美法仑具有联合作用的现象。有趣的是,使用低剂量的mTOR抑制剂,我们观察到细胞活力部分地从EPZ015938的作用中得到挽救,这表明mTOR相关途径在EPZ015938的抗骨髓瘤活性中起作用。此外,PRMT5被证明参与MMSET和SLAMF7的剪接调节,这两个基因在MM疾病中具有重要意义。因此,我们拓宽了对PRMT5在MM疾病中确切作用的理解,并进一步强调了其作为可能治疗靶点的用途。