Data Sheet 3_Network analysis of master regulators associated with invasive phenotypes in multiple myeloma.pdf

To elucidate the role of transcriptional regulators (TRs) associated with invasiveness in multiple myeloma (MM), we conducted a systematic network analysis to identify key master regulators (MRs) that govern MM invasiveness. We employed a consensus clustering method based on a 24-gene signature to classify MM patients into high invasiveness (INV-H) and low invasiveness (INV-L) groups. Subsequently, we identified TRs specific to the INV-H and INV-L phenotypes as MRs using a network-based approach, and we validated the MR activities that correlated with the INV-H phenotype across multiple independent datasets. We evaluated the effect of MRs on patient outcomes in relation to the prognosis of MM. By utilizing siRNA to disrupt ERG expression in U266 and RPMI8226 cell lines, we evaluated the effects of the master regulator ERG on the proliferation, apoptosis, invasion, and migration of myeloma cell lines, and we confirmed the expression of ERG in patients with extramedullary MM. We assessed invasiveness using a 24-gene signature, categorizing patients into INV-H and INV-L groups. Our network identified MRs linked to MM invasiveness and revealed enriched signaling pathways. High ERG expression correlated with poor prognosis. ERG silencing reduced cell invasiveness, migration, and apoptosis, while promoting proliferation. Elevated ERG was found in extramedullary MM, and potential drug candidates, including Idarubicin, were identified for treatment. This study provides a comprehensive analysis of master regulators in EMM, contributing to targeted therapeutic strategies. We identified ERG as a marker for extramedullary invasion in MM, suggesting it as a potential therapeutic target for future interventions.为阐明与多发性骨髓瘤（multiple myeloma, MM）侵袭性相关的转录调控因子（transcriptional regulators, TRs）的作用，我们开展了系统性网络分析，以鉴定调控MM侵袭性的关键主调控因子（master regulators, MRs）。我们采用基于24基因特征的共识聚类方法，将MM患者划分为高侵袭性组（INV-H）与低侵袭性组（INV-L）。随后，我们通过基于网络的方法，鉴定出与INV-H、INV-L表型特异性相关的TRs作为MRs，并在多个独立数据集中共验证了与INV-H表型相关的MR活性。我们评估了MRs对MM患者预后的影响。通过使用小干扰RNA（small interfering RNA, siRNA）干扰U266和RPMI8226细胞系中ERG的表达，我们探究了主调控因子ERG对骨髓瘤细胞系增殖、凋亡、侵袭及迁移的影响，并验证了ERG在髓外多发性骨髓瘤（extramedullary multiple myeloma, EMM）患者中的表达情况。我们基于24基因特征评估侵袭性，将患者划分为INV-H与INV-L组。我们的网络分析鉴定出与MM侵袭性相关的MRs，并揭示了富集的信号通路。高ERG表达与不良预后相关。ERG沉默可降低细胞侵袭、迁移能力并减少凋亡，同时促进细胞增殖。髓外MM患者中ERG表达升高，我们还鉴定出包括伊达比星（Idarubicin）在内的潜在治疗候选药物。本研究对EMM中的主调控因子进行了全面分析，为靶向治疗策略提供了依据。我们鉴定出ERG可作为MM髓外侵袭的标志物，提示其可作为未来干预的潜在治疗靶点。