CAS OpenIR
帕金森病靶向输递体系用于基因-化药协同治疗的研究
刘霖颖
Thesis Advisor张欣
2020-07-01
Degree Grantor中国科学院大学
Degree Name博士
Degree Discipline生物化工
Keyword帕金森病,Sirna,联合递送,仿生设计
Abstract

对于以帕金森病为典型的神经退行性疾病,目前治疗方法的瓶颈在于无法缓解多巴胺能神经元退行性病变进程。对于有望逆转神经元退行性病变的药物如基因药物和化学药物,其脑部递送存在以下共性关键科学问题:单一效果不佳,药物组织渗透差,病灶富集低,可控释药难以及难以示踪。现有用于脑疾病研究的纳米药物主要依赖于血脑屏障(BBB)的简单渗透或靶向神经元(例如使用CPP和其他肽)的开发。然而,由于大脑生理机能复杂且药物传递障碍是连续的,因此这些单一的传递步骤解决方案通常不具有高效性。因此,通过合理设计实现脑部组织的高效药物富集是脑部疾病药物递送的难点。基于此,我们构建了一种CT可视化金纳米颗粒基因-化药联合递送体系。该体系可实现药物脑部高效富集和协同治疗,具体内容如下:(1)该体系实现了药物的级联靶向:通过细胞穿透肽B6介导纳米颗粒通过血脑屏障渗透进入脑部组织,马吲哚介导纳米颗粒靶向神经元,显著提高药物在病灶的富集。(2)该体系实现了药物的可控释放:利用载体中硫醚键在H2O2响应下释放药物,实现载体在神经元内可控释药。(3)该体系实现了药物输递过程的可视化:在该Fe3+响应性金纳米颗粒造影剂内核的作用下,患病小鼠脑部CT区域有信号增强,提示纳米颗粒在脑部富集。在治疗效果方面,该联合给药纳米颗粒可发挥协同作用,有效改善帕金森病小鼠运动行为学特征,尤其在改善小鼠脑部黑质区多巴胺能神经元α-Syn蛋白和恢复小鼠脑部黑质区神经元数目等方面具有显著性作用。因此,该可视化靶向纳米系统可为缓解多巴胺能神经元退行性病变提供载体平台。针对现有传统合成递送系统存在天然免疫应答和入胞方式不佳等问题,外泌体因其血液循环的免疫惰性和膜融合入胞等优势被应用于脑部疾病的药物递送,但现有研究中外泌体难以实现将基因药物和化学药物同时高效递送至脑部病灶。因此,利用天然载体实现其携载药物在脑部组织的高效药物富集是脑部疾病药物递送的难点。本课题构建了外泌体涂层聚合物杂化联合递送纳米颗粒用于基因药物siSNCA和姜黄素协同治疗。主要方法由合成和组装过氧化氢响应的基因-化药聚合物载体,外泌体的分离和靶向多肽修饰以及复合物组装三部分组成。该体系可实现药物脑部高效富集,具体内容如下:(1)该体系实现了药物的脑部组织渗透和病灶富集:通过外泌体表面修饰的靶向多肽RVG介导纳米颗粒通过血脑屏障渗透进入脑部组织并靶向神经元,显著提高药物在病灶的富集;(2)该体系实现了药物的可控释放:利用外泌体膜融合作用,改善双载药聚合物内吞方式由内涵体途径转为膜融合途径,释放聚合物双载药内核于细胞质,载体内核中苯硼酸基团在H2O2响应下释放药物,实现载体在神经元内可控释药。在治疗效果方面:(1)上述高效递送优势可增强siSNCA和姜黄素协同治疗效果。(2)该体系在多种动物帕金森病模型治疗中获得良好效果:分别在MPTP诱导的帕金森病小鼠,纹状体内单侧注射α-Syn寡聚体帕金森病大鼠,纹状体内单侧注射α-Syn寡聚体帕金森病食蟹猴模型行为学改善中均发挥了积极作用。(3)本研究初步证实了该体系对T细胞免疫抑制作用,其可为研究未成熟树突状外泌体载体系统的治疗提供新思路。因此,本课题基于基因药物siSNCA和化学药物姜黄素对α-突触核蛋白聚集体清除的潜在协同作用,围绕基因-化学药物联合递送策略展开,构建了两种靶向递送体系:CT可视化金纳米颗粒基因-化药联合递送体系和外泌体涂层聚合物杂化联合递送纳米颗粒,实现了脑部疾病药物递送的联合给药,高药物组织渗透,高病灶富集,可控释药和可视化递送,解决了基因和化药在脑部的高效富集的难点,缓解了神经元退行性病变,为帕金森病靶向治疗提供前瞻性方案。;For the neurodegenerative disease such as Parkinson's disease, the bottleneck of current treatment is that it can’t alleviate the dopaminergic neuronal degeneration. Considering the key pathological of Parkinson's disease, α-synuclein (α-Syn), we proposes a strategy of synergistic treatment with gene drugs and chemical drugs. There are a series of problems in the drug delivery process, such as poor tissue permeability, low lesion concentration, difficulty of drug controlled release and difficulty to track.Existing nanomedicines currently used for brain disease research rely primarily on the simple penetration of the blood-brain barrier (BBB) or the development of targeted neurons delivery, such as using CPP and other peptides. However, brain physiological functions are complex and drug delivery disorders are continuous, these single delivery step solutions are often not efficient. Therefore, efficient drug enrichment of brain tissue through rational design is a difficult point in drug delivery for brain diseases. Based on this, we constructed a CT visualized gold nanoparticle gene-chemical drug delivery system. The system can achieve efficient brain drug enrichment and synergistic therapy. The specific contents are as follows: (1) The system achieves cascade targeting of drugs: through cell-penetrating peptide B6, the nanoparticles are permeable to the brain through the blood-brain barrier, and achieves neurons target via mazindol which significantly increasing drug accumulation in the lesion. (2) The system achieves drug controlled release: thioether bonds in the carrier release the drug in response to H2O2, to achieve drug controlled release in the neuron. (3) The system realizes the visualization of drug delivery process: under the action of the Fe3+ responsive gold nanoparticle contrast agent core, there is a CT signal enhancement in the the brain of diseased mice, suggesting that the nanoparticles are enriched in the brain. In terms of therapeutic effect, the combined delivery of gene and chemical drugs can play a synergistic effect and effectively improve the motor behavioral characteristics of Parkinson's disease mice, especially in decreasing the dopaminergic neuron α-Syn protein in the brain's substantia nigra region and recovering the number of neurons in the substantia nigra area. Therefore, the visual targeting nanosystem can provide a carrier platform for relieving dopaminergic neuron degeneration.Aiming at the problems of the existing traditional synthetic delivery system, such as natural immune response and poor cell uptake, exosome has been applied to drug delivery for brain diseases due to their advantages such as immune inertia in blood circulation and membrane fusion into cells. However, it is difficult to achieve efficient and simultaneous delivery of gene drugs and chemical drugs to brain lesions in the existing research. Therefore, exploring the combined delivery of targeted natural carriers for efficient drug enrichment in brain tissue is a difficult point in drug delivery for brain diseases. We proposes a strategy of exosome coating polymer hybrid nanoparticle as combined drug delivery system for the synergistic treatment in order to enhance drug delivery efficiency. The main method consists of the synthesis and assembly of a hydrogen peroxide-responsive gene-chemical drug polymer carrier, the isolation of exosomes, the modification of targeted polypeptides on exosome and the assembly of complexes. We confirms the efficient drug delivery: (1) The system achieves brain tissue penetration and lesion enrichment of the drug: the RVG modified exosome mediates nanoparticles across the blood-brain barrier, penetrate into brain tissue and target neurons, which significantly increase drug accumulation in the lesion. (2) The system achieves controlled release of the drug: exosome membrane fuses with cells to improve the dual-drug polymer endocytosis. The endosome pathway is changed to the membrane fusion pathway, and the polymer based drug core is released into the cytoplasm. The phenylboronic acid group in the carrier core release the drug in response to H2O2. In terms of therapeutic effects: (1) The above-mentioned efficient delivery advantages can enhance the synergistic therapeutic effect of siSNCA and curcumin. (2) The system results in the treatment of various animal Parkinson's disease models: MPTP-induced Parkinson's disease mice, unilateral striatum α-Syn oligomers injected Parkinson's disease rats and monkeys. (3) This study has confirmed the immunosuppressive effect of this system on T cells, which can provide new ideas for the application of immature dendritic exosome systems.Therefore, based on the potential synergistic effect of siSNCA and curcumin on the clearance of α-synuclein aggregates, we construct two gene-chemical drug delivery systems: CT visualization gold nanoparticle for gene-chemical drug delivery system and exosome coating polymer co-delivery hybrid system. The systems are combined drug delivery for brain diseases, have high drug tissue penetration, high lesion enrichment, controlled drug release and track ability. They have solved the difficulty of efficient enrichment of genes and chemical drugs in the brain, have relieved neuron degeneration and provide a prospective solution for targeted treatment of Parkinson's disease. 

Language中文
Document Type学位论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/49658
Collection中国科学院过程工程研究所
Recommended Citation
GB/T 7714
刘霖颖. 帕金森病靶向输递体系用于基因-化药协同治疗的研究[D]. 中国科学院大学,2020.
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