Han Jang Mi, Kim Yu Jin, Jung Hye Jin
Department of Pharmaceutical Engineering & Biotechnology, Sun Moon University, Asan 31460, Korea.
Genome-Based BioIT Convergence Institute, Sun Moon University, Asan 31460, Korea.
Cancers (Basel). 2022 Mar 4;14(5):1315. doi: 10.3390/cancers14051315.
Glioblastoma stem-like cells (GSCs) drive tumor initiation, cancer invasion, immune evasion, and therapeutic resistance and are thus a key therapeutic target for improving treatment for glioblastoma multiforme (GBM). We previously identified calcium/calmodulin-dependent protein kinase II (CaMKII) as an emerging molecular target for eliminating GSCs. In this study, we aim to explore a new CaMKII-targeted synthetic lethal therapy for GSCs. Through high-throughput drug combination screening using CaMKII inhibitors and a bioactive compound library in GSCs, neurokinin 1 receptor (NK1R) inhibitors such as SR 140333 and aprepitant are found to be potential anticancer agents that exhibit chemical synthetic lethal interactions with CaMKII inhibitors, including hydrazinobenzoylcurcumin (HBC), berbamine, and KN93. Combined treatment with NK1R and CaMKII inhibitors markedly suppresses the viability and neurosphere formation of U87MG- and U373MG-derived GSCs. In addition, the combination of HBC and NK1R inhibitors significantly inhibits U87MG GSC tumor growth in a chick embryo chorioallantoic membrane (CAM) model. Furthermore, the synthetic lethal interaction is validated using RNA interference of CaMKIIγ and NK1R. Notably, the synthetic lethal effects in GSCs are associated with the activation of caspase-mediated apoptosis by inducing p53 expression and reactive oxygen species generation, as well as the suppression of stemness marker expression by reducing nuclear factor-kappa B (NF-κB) activity. This follows the downregulation of phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling and a decrease in intracellular calcium concentration. Moreover, NK1R affects CaMKIIγ activation. These findings demonstrate that NK1R is a potential synthetic lethal partner of CaMKII that is involved in eradicating GSCs, and they suggest a new CaMKII-targeted combination therapy for treating GBM.
胶质母细胞瘤干细胞（GSCs）驱动肿瘤起始、癌症侵袭、免疫逃逸和治疗抵抗，因此是改善多形性胶质母细胞瘤（GBM）治疗的关键治疗靶点。我们之前将钙/钙调蛋白依赖性蛋白激酶II（CaMKII）鉴定为消除GSCs的一个新出现的分子靶点。在本研究中，我们旨在探索一种针对GSCs的新型CaMKII靶向合成致死疗法。通过在GSCs中使用CaMKII抑制剂和生物活性化合物库进行高通量药物组合筛选，发现神经激肽1受体（NK1R）抑制剂如SR 140333和阿瑞匹坦是潜在的抗癌药物，它们与包括肼基苯甲酰姜黄素（HBC）、小檗胺和KN93在内的CaMKII抑制剂表现出化学合成致死相互作用。NK1R和CaMKII抑制剂联合治疗显著抑制U87MG和U373MG来源的GSCs的活力和神经球形成。此外，HBC和NK1R抑制剂的组合在鸡胚绒毛尿囊膜（CAM）模型中显著抑制U87MG GSC肿瘤生长。此外，使用CaMKIIγ和NK1R的RNA干扰验证了合成致死相互作用。值得注意的是，GSCs中的合成致死效应与通过诱导p53表达和活性氧生成激活半胱天冬酶介导的凋亡有关，以及通过降低核因子-κB（NF-κB）活性抑制干性标志物表达有关。这伴随着磷酸肌醇3激酶（PI3K）/蛋白激酶B（AKT）信号通路的下调和细胞内钙浓度的降低。此外，NK1R影响CaMKIIγ的激活。这些发现表明NK1R是CaMKII的潜在合成致死伙伴，参与消除GSCs，并且它们提示了一种用于治疗GBM的新型CaMKII靶向联合疗法。
