Combined MEK and JAK/STAT3 pathway inhibition effectively decreases medulloblastoma tumor progression.

RNA sequencing was carried out to identify the signaling pathways and hallmarks that change following MEK and/or JAK/STAT3 pathway inhibition. Overall design: (1) 2.5x10^5 UI226 cells were injected into the cerebellums of NOD SCID mice. Mice were treated with either vehicle controls of selumetinib twice daily (drug holiday on the weekend) via oral gavage. Tumor were extracted at endpoint and human cells were isolated by FACS with HLA. Human cells from vehicle and selumetinib treated xenografts were subject to RNA-seq. (2) We performed RNA sequencing on selumetinib and/or pacritinib treated UI226 tumorspheres (N=4-6 biological replicates for each condition) to characterize the sustained transcriptomic changes 5 days following treatment in vitro. Tumorspheres were treated with vehicle, selumetinib only, pacritinib only or selumetinib + pacritinib

本研究通过RNA测序(RNA sequencing)，旨在鉴定MEK及/或JAK/STAT3通路抑制后发生改变的信号通路与特征标志物。整体实验设计如下：1. 将2.5×10^5个UI226细胞接种至NOD/SCID小鼠的小脑内。小鼠通过灌胃给药，分别给予溶剂对照或每日两次的司美替尼(selumetinib，周末设置药物假期)。于实验终点摘取肿瘤组织，通过针对人类白细胞抗原(HLA)的流式细胞术(FACS)分离得到人源细胞；随后对溶剂对照及司美替尼处理的异种移植瘤来源的人源细胞进行RNA测序(RNA-seq)。2. 本研究对经司美替尼、帕瑞替尼(pacritinib)单药或联合处理的UI226肿瘤球开展RNA测序，每组设置4~6个生物学重复，以表征体外给药5天后持续存在的转录组学改变。实验分组涵盖溶剂对照、单药司美替尼、单药帕瑞替尼以及司美替尼联合帕瑞替尼。