Riess Christin, Koczan Dirk, Schneider Björn, Linke Charlotte, Del Moral Katharina, Classen Carl Friedrich, Maletzki Claudia
University Children's Hospital, Rostock University Medical Centre, Ernst-Heydemann-Straße 8, 18057, Rostock, Germany.
Department of Medicine Clinic III - Hematology, Oncology, Palliative Medicine, Rostock University Medical Center, Rostock University Medical Centre, Ernst-Heydemann-Str. 6, 18057, Rostock, Germany.
Cell Death Discov. 2021 Mar 15;7(1):54. doi: 10.1038/s41420-021-00423-1.
Current therapeutic approaches have met limited clinical success for glioblastoma multiforme (GBM). Since GBM harbors genomic alterations in cyclin-dependent kinases (CDKs), targeting these structures with specific inhibitors (CDKis) is promising. Here, we describe the antitumoral potential of selective CDKi on low-passage GBM 2D- and 3D models, cultured as neurospheres (NSCs) or glioma stem-like cells (GSCs). By applying selective CDK4/6i abemaciclib and palbociclib, and the more global CDK1/2/5/9-i dinaciclib, different effects were seen. Abemaciclib and dinaciclib significantly affected viability in 2D- and 3D models with clearly visible changes in morphology. Palbociclib had weaker and cell line-specific effects. Motility and invasion were highly affected. Abemaciclib and dinaciclib additionally induced senescence. Also, mitochondrial dysfunction and generation of mitochondrial reactive oxygen species (ROS) were seen. While autophagy was predominantly visible after abemaciclib treatment, dinaciclib evoked γ-H2AX-positive double-strand breaks that were boosted by radiation. Notably, dual administration of dinaciclib and abemaciclib yielded synergistic effects in most cases, but the simultaneous combination with standard chemotherapeutic agent temozolomide (TMZ) was antagonistic. RNA-based microarray analysis showed that gene expression was significantly altered by dinaciclib: genes involved in cell-cycle regulation (different CDKs and their cyclins, SMC3), mitosis (PLK1, TTK), transcription regulation (IRX3, MEN1), cell migration/division (BCAR1), and E3 ubiquitination ligases (RBBP6, FBXO32) were downregulated, whereas upregulation was seen in genes mediating chemotaxis (CXCL8, IL6, CCL2), and DNA-damage or stress (EGR1, ARC, GADD45A/B). In a long-term experiment, resistance development was seen in 1/5 cases treated with dinaciclib, but this could be prevented by abemaciclib. Vice versa, adding TMZ abrogated therapeutic effects of dinaciclib and growth was comparable to controls. With this comprehensive analysis, we confirm the therapeutic activity of selective CDKi in GBM. In addition to the careful selection of individual drugs, the timing of each combination partner needs to be considered to prevent resistance.
目前的治疗方法在多形性胶质母细胞瘤（GBM）的临床治疗中取得的成功有限。由于GBM在细胞周期蛋白依赖性激酶（CDK）中存在基因组改变，用特异性抑制剂（CDKis）靶向这些结构具有前景。在此，我们描述了选择性CDKi对低传代GBM二维和三维模型的抗肿瘤潜力，这些模型培养为神经球（NSCs）或胶质瘤干细胞样细胞（GSCs）。通过应用选择性CDK4/6抑制剂阿贝西利和哌柏西利，以及更具全局性的CDK1/2/5/9抑制剂地西他滨，观察到了不同的效果。阿贝西利和地西他滨显著影响二维和三维模型中的细胞活力，形态学上有明显可见的变化。哌柏西利的作用较弱且具有细胞系特异性。迁移和侵袭受到高度影响。阿贝西利和地西他滨还诱导了衰老。此外，还观察到线粒体功能障碍和线粒体活性氧（ROS）的产生。虽然阿贝西利治疗后自噬主要可见，但地西他滨诱发了γ-H2AX阳性双链断裂，辐射可增强这种断裂。值得注意的是，在大多数情况下，地西他滨和阿贝西利联合给药产生协同效应，但与标准化疗药物替莫唑胺（TMZ）同时联合则具有拮抗作用。基于RNA的微阵列分析表明，地西他滨显著改变了基因表达：参与细胞周期调控（不同的CDK及其细胞周期蛋白、SMC3）、有丝分裂（PLK1、TTK）、转录调控（IRX3、MEN1）、细胞迁移/分裂（BCAR1）和E3泛素化连接酶（RBBP6、FBXO32）的基因下调，而介导趋化作用（CXCL8、IL6、CCL2）以及DNA损伤或应激（EGR1、ARC、GADD45A/B）的基因上调。在一项长期实验中，接受地西他滨治疗的5例患者中有1例出现耐药，但阿贝西利可预防这种情况。反之，添加TMZ会消除地西他滨的治疗效果，生长情况与对照组相当。通过这项全面分析，我们证实了选择性CDKi在GBM中的治疗活性。除了仔细选择个体药物外，还需要考虑每个联合用药伙伴的给药时机以预防耐药。
