基于自噬抑制的靶向多功能纳米粒子的构建及其在脑胶质瘤治疗中的应用与机制研究

Enrichment of brain-targeting chemo-therapeutic drug in the brain will improve the curative effect of treating brain glioma. But, most of chemo-therapeutic drug can produce protective autophagy simultaneously and will induce the glioma cell drug resistance and decrese the effective drug delivery. To further investigate the autophagy caused by chemo-therapeutic drug and the mechanism of Inhibition autophagy, enhance drug delivery and decrese drug resistance, we constructed multifunction brain-targeting paclitaxel nanoparticles (NPs). We plan to study the multifunction paclitaxel nanoparticles (NPs) inhibit glioma cells. Acoording to the process of autophagy role, taking a key preotein Beclin-1, ATG5 and ATG7 in the autophagy as a targeting. Also, we will seek for the miRNA that inhibit protective autophagy of glioma cell, synthesis this miRNA, and construct the co-dilivery of paclitaxel , miRNA and autophagy inhibitors multifuctional targeting nano-carrier system. Use the way of brain-targeting of NPs to verify the effect on passing through the blood brain barrier. We will detect the enrichment and the localization of drug-loaded NPs in glioma, analysis the effect of drug-loaded NPs on suppress glioma autophagy, and the efficacy of arousing glioma cell apoptosis. We will preliminary clarify the regulatory role of miRNA combinate paclitaxel and autophagy inhibitors in inhibition autophagy aroused by chemo-therapeutic drug, preliminary study the pathway involved in the mechanism of autophagy in tumor resistance and achieve the treatment of the glioma by the co-delivery of chemotherapy drug and gene brain-targeting NPs, which will provide new targets and ideas for the curative effect glioma .

脑靶向化疗药物能在脑内富集，提高了对脑胶质瘤的治疗疗效，但多数化疗药物能同时产生保护性自噬引起胶质瘤细胞耐药，降低了有效的药物递送。为了进一步研究化疗药物引起的自噬，研究抑制自噬提高药物递送及降低耐药的机制，我们构建了多功能脑靶向紫杉醇纳米粒子并研究其抑制胶质瘤的效果。以自噬途径中的关键蛋白Beclin-1、ATG5和ATG7为靶点，寻找抑制胶质瘤细胞保护性自噬的miRNA，合成miRNA并构建同时携载紫杉醇、miRNA及自噬抑制剂的靶向纳米载药系统，验证其透过血脑屏障作用，检测其在脑胶质瘤内的富集与定位，分析其抑制胶质瘤细胞自噬的作用、引起胶质瘤细胞凋亡药效，初步阐明miRNA联合紫杉醇、自噬抑制剂在抑制化疗药物引起的自噬反应中的调控作用，并对自噬作用在肿瘤耐药性方面的机制所涉及的通路进行初步研究，实现化疗与基因药物联合构建靶向纳米粒子治疗脑胶质瘤，为脑胶质瘤的的治疗提供新的靶点和思路。

脑靶向化疗药物能在脑内富集，提高了对脑胶质瘤的治疗疗效，但多数化疗药物能同时产生保护性自噬引起胶质瘤细胞耐药，降低了有效的药物递送。我们成功构建了能同时携载PTX、氯喹(CQ)、3-甲基腺嘌呤（3-MA）的基于葡聚糖、聚三亚甲基碳酸酯和聚乙烯亚胺的靶向纳米粒子；对紫杉醇(PTX) 引起的脑胶质瘤细胞自噬反应与耐药性相关性及miRNA异常表达谱进行了分析研究；筛选并构建了携载PTX的纳米粒子；进行了抗胶质瘤药效及抑制胶质瘤细胞自噬的效应分析研究；筛选并构建了同时携载PTX、CQ、3-MA的靶向纳米粒子；对抗脑胶质瘤药效及抑制胶质瘤细胞自噬水平进行了分析研究；筛选出以自噬相关基因Beclin-1、ATG5及ATG7等为靶点的miRNA，并对通过调节异常表达miRNA水平对自噬水平进行了调控研究；构建了同时携载PTX、CQ、3-MA、miRNA和乳铁蛋白（Lf）的纳米粒子；对抗脑胶质瘤药效及抑制胶质瘤细胞自噬水平进行了分析研究；对细胞和动物模型中的自噬通路进行分析研究，对各种自噬因子对肿瘤细胞耐药性的贡献进行评价研究。为脑胶质瘤的的治疗提供了新思路。后续研究正在进行负载PTX、CQ及miRNA多功能纳米载药粒子抗脑胶质瘤干细胞的相关研究工作。发表相关的SCI 文章9篇，授权专利3 项。