如何准确指导胶质母细胞瘤（GBM）患者选择合适化疗方案，是基础研究与临床工作的重点。NF1基因突变虽然被用来鉴别GBM分型，但尚不清楚是否通过调控血管周细胞（PC）募集改善化疗效果。我们前期发现GBM细胞NF1基因突变抑制PC增殖、血管生成以及提高HDAC6抑制剂敏感性。基于此，本项目拟将转基因斑马鱼作为模式动物，基于慢病毒转染、CRISPR/Cas9基因编辑和显微注射构建NF1基因突变的GBM原位移植瘤模型，采用激光共聚焦显微镜进行时间延迟成像，活体观察NF1基因突变对GBM细胞动态归巢和PC募集过程的影响，并检测PDGFB-PDGFRβ信号通路和HDAC6改变情况。同时，构建体外血肿瘤屏障（BTB）模型和裸鼠原位移植瘤模型，分析NF1基因突变对BTB选择性开放及对HDAC6抑制剂化疗效果的调节。本项目将揭示NF1基因突变提高GBM化疗敏感性的分子机制，为精准化疗提供新靶点和新策略。

The focus on basic research and clinical treatment is how to accurately guide glioblastoma (GBM) patients to choose better chemotherapy. Although the mutation of NF1 gene determines GBM subtype, it is not clear whether it improves chemotherapy effect by regulating the pericyte (PC) recruitment. Our previous results showed that NF1 gene mutation in GBM cells inhibited PC proliferation, angiogenesis and increased sensitivity to HDAC6 inhibitors. This project is to construct orthotopic transplantation model of GBM with NF1 gene mutation by means of the technology offluorescent lentivirus transfection, CRISPR/Cas9 and microinjection based on transgenic zebrafish. Combining with laser confocal microscopy, the time lapse imaging technique was used to observe the influence of NF1 gene mutation on GBM cells dynamic homing and PC recruitment in vivo. The expression changes of PDGFB-PDGFRβ signaling pathway and HDAC6 will be tested. Meanwhile, blood tumor barrier (BTB) and orthotropic transplantation tumor model in nude mice will be established to analyze the effect of NF1 gene mutation on selective opening of BTB and chemotherapy effect of HDAC6 inhibitors. This subject will reveal the molecular mechanism of NF1 gene mutation for improving GBM chemotherapy sensitivity, and provide new targets and strategies for precise chemotherapy.