组蛋白去乙酰化酶（HDAC）与赖氨酸特异性去甲基化酶1A（KDM1A）联合抑制在胶质母细胞瘤中的临床前活性
Preclinical activity of combined HDAC and KDM1A inhibition in glioblastoma.
作者信息
Singh Melissa M, Johnson Blake, Venkatarayan Avinashnarayan, Flores Elsa R, Zhang Jianping, Su Xiaoping, Barton Michelle, Lang Frederick, Chandra Joya
机构信息
Department of Pediatrics Research, University of Texas MD Anderson Cancer Center, Houston, Texas (M.M.S., B.J., J.C.); Department of Molecular and Cellular Oncology, University of Texas MD Anderson Cancer Center, Houston, Texas (A.V., E.R.F.); Department of Molecular Carcinogenesis, University of Texas MD Anderson Cancer Center, Houston, Texas (M.B., J.C.); Department of Bioinformatics and Computational Biology, University of Texas MD Anderson Cancer Center, Houston, Texas (J.Z., X.S.); Department of Neurosurgery, University of Texas MD Anderson Cancer Center, Houston, Texas (B.J., F.L.); Graduate School of Biomedical Sciences, University of Texas Health Science Center, Houston, Texas (A.V., E.R.F., M.B., F.L., J.C.).
出版信息
Neuro Oncol. 2015 Nov;17(11):1463-73. doi: 10.1093/neuonc/nov041. Epub 2015 Mar 19.
BACKGROUND
Glioblastoma (GBM) is the most common and aggressive form of brain cancer. Our previous studies demonstrated that combined inhibition of HDAC and KDM1A increases apoptotic cell death in vitro. However, whether this combination also increases death of the glioma stem cell (GSC) population or has an effect in vivo is yet to be determined. Therefore, we evaluated the translational potential of combined HDAC and KDM1A inhibition on patient-derived GSCs and xenograft GBM mouse models. We also investigated the changes in transcriptional programing induced by the combination in an effort to understand the induced molecular mechanisms of GBM cell death.
METHODS
Patient-derived GSCs were treated with the combination of vorinostat, a pan-HDAC inhibitor, and tranylcypromine, a KDM1A inhibitor, and viability was measured. To characterize transcriptional profiles associated with cell death, we used RNA-Seq and validated gene changes by RT-qPCR and protein expression via Western blot. Apoptosis was measured using DNA fragmentation assays. Orthotopic xenograft studies were conducted to evaluate the effects of the combination on tumorigenesis and to validate gene changes in vivo.
RESULTS
The combination of vorinostat and tranylcypromine reduced GSC viability and displayed efficacy in the U87 xenograft model. Additionally, the combination led to changes in apoptosis-related genes, particularly TP53 and TP73 in vitro and in vivo.
CONCLUSIONS
These data support targeting HDACs and KDM1A in combination as a strategy for GBM and identifies TP53 and TP73 as being altered in response to treatment.
背景
胶质母细胞瘤（GBM）是最常见且侵袭性最强的脑癌形式。我们之前的研究表明，联合抑制组蛋白去乙酰化酶（HDAC）和赖氨酸特异性去甲基化酶1A（KDM1A）可在体外增加凋亡细胞死亡。然而，这种联合治疗是否也能增加胶质瘤干细胞（GSC）群体的死亡或在体内产生效果尚待确定。因此，我们评估了联合抑制HDAC和KDM1A对患者来源的GSC和异种移植GBM小鼠模型的转化潜力。我们还研究了联合治疗诱导的转录程序变化，以了解GBM细胞死亡的诱导分子机制。
方法
用泛HDAC抑制剂伏立诺他和KDM1A抑制剂反苯环丙胺联合处理患者来源的GSC，并检测其活力。为了表征与细胞死亡相关的转录谱，我们使用RNA测序（RNA-Seq），并通过逆转录定量聚合酶链反应（RT-qPCR）验证基因变化，通过蛋白质免疫印迹法验证蛋白质表达。使用DNA片段化分析检测细胞凋亡。进行原位异种移植研究，以评估联合治疗对肿瘤发生的影响，并在体内验证基因变化。
结果
伏立诺他和反苯环丙胺的联合使用降低了GSC的活力，并在U87异种移植模型中显示出疗效。此外，联合治疗在体外和体内均导致凋亡相关基因的变化，特别是TP53和TP73。
结论
这些数据支持联合靶向HDAC和KDM1A作为GBM的一种治疗策略，并确定TP53和TP73会因治疗而发生改变。
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