Effect of AsiDNA and belinostat on gene expression in U87 cells. [RNA-Seq]. Effect of AsiDNA and belinostat on gene expression in U87 cells. [RNA-Seq]

An effective long-term inhibition of multiple DNA repair signals is required to design a novel and effective therapeutic for glioblastoma (GBM), a highly DNA repair ‘addicted’ cancer. AsiDNA, a double-strand DNA break (DSB) mimetic, is an innovative approach that shuts down the entire DNA repair system in cancer cells by sequestering DSB repair factors. We showed that inhibition of class I histone deacetylases (HDACs) dislodges repair factors from sites of DNA damage. We therefore tested whether AsiDNA and pan HDAC inhibitor Belinostat combination can impart a chromatin-based long-term DNA damage and impair DNA repair in GBM cells. In order to understand the long-term effects of AsiDNA on DNA repair, we treated U87 cells with AsiDNA for 6 days followed by 6 days recovery from the drug. We used this protocol to create long-term AsiDNA treated cells (LTAU87) according to the clinical trial protocol. We performed RNA-Seq with LTAU87 cells and MTU87 (mock treated U87; control) in the presence and absence of belinostat. Overall design: Comparative gene expression analysis was performed following exposure of glioblastoma cells to AsiDNA and belinostat.

开发针对胶质母细胞瘤（glioblastoma, GBM）——一种高度依赖DNA修复的"成瘾性"癌症——的新型有效治疗手段，需实现对多种DNA修复信号的长效、强效抑制。AsiDNA作为双链DNA断裂（double-strand DNA break, DSB）模拟剂，是一种创新策略：它通过螯合DSB修复因子，关闭癌细胞内的整套DNA修复系统。我们的研究证实，I类组蛋白去乙酰化酶（class I histone deacetylases, HDACs）的抑制可将修复因子从DNA损伤位点剥离。基于此，我们检验了AsiDNA与广谱HDAC抑制剂贝利司他（Belinostat）联合使用，能否在胶质母细胞瘤细胞中诱导基于染色质的长期DNA损伤，并削弱其DNA修复能力。为探究AsiDNA对DNA修复的长期影响，我们将U87细胞用AsiDNA连续处理6天，随后停药恢复6天，并参照临床试验方案构建了长期AsiDNA处理的U87细胞系（LTAU87）。我们分别在贝利司他存在与不存在的条件下，对LTAU87细胞与MTU87细胞（即Mock处理的U87细胞，作为对照）开展RNA测序（RNA-Seq）。整体实验设计：对经AsiDNA与贝利司他处理的胶质母细胞瘤细胞进行比较基因表达分析。