胶质母细胞瘤诊疗一体CdTe/ZnS量子点靶向纳米探针研究

由于胶质母细胞瘤（GBM）边界不清，患者存在肿瘤切除不全和复发的风险，所以识别肿瘤边界是手术的关键。申请人前期研究发现CdTe/ZnS 量子点（QDs），粒径小，荧光强，可经尾静脉进入小鼠脑组织成像，在成像浓度下对小鼠无明显毒效应，适合制备GBM靶向探针；另外，CdTe/ZnS QDs还可激活小胶质细胞mTOR通路、NLRP3炎症小体，促使细胞向M1型分化，具有GBM免疫治疗潜力。因此，本项目拟：①构建GBM靶向探针：电化学制备CdTe/ZnS QDs，表面羧基连接EGFR（vIII）抗体并表征。②验证探针靶向性：构建体外血脑屏障模型验证探针靶向性；构建GBM裸鼠模型，分析探针体内成像、代谢和毒性结果。③免疫治疗作用验证：小鼠体内、体外分析探针对小胶质M1型极化水平的影响及机制，分析探针对荷瘤小鼠肿瘤尺寸、生存曲线的影响。最终，综合评价该探针在GBM诊疗方面的价值。

Glioblastomas(GBM) have a high recurrence rate because the tumor boundaries are unclear and surgical resection rates are usually low. Precise identification of tumor boundaries is the key to the surgery. Applicant's previous study found that CdTe/ZnS quantum dots (QDs) have small particle size but strong fluorescence. They successfully permeated into the mouse brain via tail vein injection, and in vivo imaged. No significant toxic effect was observed on mice at the imaging concentration. So CdTe/ZnS QD is an ideal material for assembling GBM probe. In addition, applicant's study found that CdTe/ZnS QDs can activate the mTOR pathway and NLRP3 inflammasome of microglia, which could polarize microglia to M1 type. So CdTe/ZnS QDs have potential for GBM immunotherapy. Based on that, this project is planed as follow: ① Prepare GBM targeting probes. CdTe/ZnS QDs are prepared by electrochemical synthesis method, and carboxyl groups on the surface are linked to EGFR (vIII) antibodies; Characterize the probe. ② Verify the targeting. Constructing the blood-brain barrier model in vitro to verify the targeting of the probe; Using GBM nude mice model to analyze the imaging, metabolism and toxicity of the probe in vivo. ③ Verify the effects of immunotherapy. In vivo and in vitro, the effects of probes on M1 type polarization of microglia will be analyzed. Analyzing the influence of probe on tumor size and survival curve of mice model. Explore the molecular mechanism of the polarization induced by the probe. Above all, this project will evaluated the value of the probe in diagnosis and treatment of GBM.