Puccio Noemi, Manzotti Gloria, Mereu Elisabetta, Torricelli Federica, Ronchetti Domenica, Cumerlato Michela, Craparotta Ilaria, Di Rito Laura, Bolis Marco, Traini Valentina, Manicardi Veronica, Fragliasso Valentina, Torrente Yvan, Amodio Nicola, Bolli Niccolò, Taiana Elisa, Ciarrocchi Alessia, Piva Roberto, Neri Antonino
Laboratory of Translational Research, Azienda USL-IRCCS di Reggio Emilia, Reggio Emilia; Clinical and Experimental Medicine PhD Program, University of Modena and Reggio Emilia, Modena.
Laboratory of Translational Research, Azienda USL-IRCCS di Reggio Emilia, Reggio Emilia.
Haematologica. 2024 Dec 1;109(12):4040-4055. doi: 10.3324/haematol.2024.285470.
Multiple myeloma (MM) is a dreadful disease, marked by the uncontrolled proliferation of clonal plasma cells within the bone marrow. It is characterized by a highly heterogeneous clinical and molecular background, supported by severe genomic alterations. Important de-regulation of long non-coding RNA (lncRNA) expression, which can influence progression and therapy resistance, has been reported in MM patients. NEAT1 is a lncRNA essential for nuclear paraspeckles and is involved in the regulation of gene expression. We showed that NEAT1 supports MM proliferation, making this lncRNA an attractive therapeutic candidate. Here, we used a combinatorial strategy integrating transcriptomic and computational approaches with functional high-throughput drug screening to identify compounds that synergize with NEAT1 inhibition in restraining MM cell growth. AURKA inhibitors were identified as top-scoring drugs in these analyses. We showed that the combination of NEAT1 silencing and AURKA inhibitors in MM profoundly impairs microtubule organization and mitotic spindle assembly, finally leading to cell death. Analysis of the large publicly available CoMMpass dataset showed that, in MM patients, AURKA expression is strongly associated with reduced progression-free survival (P<0.0001) and overall survival (P<0.0001) probabilities and patients with high levels of expression of both NEAT1 and AURKA have a worse clinical outcome. Finally, using RNA-sequencing data from NEAT1 knockdown MM cells, we identified the AURKA allosteric regulator TPX2 as a new NEAT1 target in MM and as a mediator of the interplay between AURKA and NEAT1, therefore providing a possible explanation for the synergistic activity observed upon their combinatorial inhibition.
多发性骨髓瘤(MM)是一种可怕的疾病,其特征是骨髓内克隆性浆细胞不受控制地增殖。它具有高度异质性的临床和分子背景,并伴有严重的基因组改变。已有报道称,MM患者中长链非编码RNA(lncRNA)表达的重要失调会影响疾病进展和治疗耐药性。NEAT1是一种对核旁斑至关重要的lncRNA,参与基因表达的调控。我们发现NEAT1支持MM的增殖,这使得这种lncRNA成为一个有吸引力的治疗靶点。在此,我们采用了一种将转录组学和计算方法与功能性高通量药物筛选相结合的策略,以鉴定在抑制MM细胞生长方面与NEAT1抑制协同作用的化合物。在这些分析中,极光激酶A(AURKA)抑制剂被确定为得分最高的药物。我们发现,在MM中,NEAT1沉默与AURKA抑制剂联合使用会严重损害微管组织和有丝分裂纺锤体组装,最终导致细胞死亡。对公开的大型CoMMpass数据集的分析表明,在MM患者中,AURKA表达与无进展生存期(P<0.0001)和总生存期(P<0.0001)概率降低密切相关,并且NEAT1和AURKA表达水平都高的患者临床结局更差。最后,利用来自NEAT1敲低的MM细胞的RNA测序数据,我们确定AURKA变构调节剂TPX2是MM中一个新的NEAT1靶点,也是AURKA与NEAT1相互作用的介质,因此为它们联合抑制时观察到的协同活性提供了一种可能的解释。