经超声磁传感基因载体实现帕金森病基因治疗的初步研究：克服小鼠体/内外屏障。

A preliminary study of Parkinson's gene therapy via sono-magnetic sensing gene vector for conquering extra/intracellular barriers in mice.

机构信息

Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu, Taiwan.

Department of Medical Science, Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu, Taiwan.

出版信息

Brain Stimul. 2020 May-Jun;13(3):786-799. doi: 10.1016/j.brs.2020.02.024. Epub 2020 Feb 24.

DOI:10.1016/j.brs.2020.02.024

PMID:32289709

Abstract

BACKGROUND

Non-virus genetic treatment for Parkinson's disease (PD) via plasmid glial cell-line derived neurotrophic factor (pGDNF) has shown potential for repairing damaged dopaminergic neurons. However, development of this gene therapy is largely hampered by the insufficient transfection efficiency as a result of the cell membrane, lysosome, and cytoskeleton meshwork.

METHODS

In this study, we propose the use of polyethylenimine (PEI)-superparamagnetic iron oxide-plasmid DNA (pDNA)-loaded microbubbles (PSp-MBs) in conjunction with focused ultrasound (FUS) and two-step magnetic navigation to provide cavitation, proton sponge effect and magnetic effects to increase the efficiency of gene delivery.

RESULTS

The gene transfection rate in the proposed system was 2.2-fold higher than that of the commercial agent (TransIT®-LT1). The transfection rate could be boosted ∼11%, ∼10%, and 6% by cavitation-magnetic hybrid enhanced cell membrane permeabilization, proton sponge effect, and magnetic-assisted cytoskeleton-reorganization, respectively. In vivo data suggested that effective gene delivery with this system results in a 3.2-fold increase in recovery of dopaminergic neurons and a 3.9-fold improvement in the motor behavior when compared to untreated genetic PD mice.

CONCLUSIONS

We proposed that this novel FUS-magnetic hybrid gene delivery platform could be integrated with a variety of therapeutic genes for treating neurodegenerative diseases in the future.

摘要

背景

通过质粒胶质细胞衍生神经营养因子（pGDNF）对帕金森病（PD）进行非病毒基因治疗显示出修复受损多巴胺能神经元的潜力。然而，由于细胞膜、溶酶体和细胞骨架网络，这种基因治疗的发展在很大程度上受到转染效率不足的阻碍。

方法

在这项研究中，我们提出使用聚乙烯亚胺（PEI）-超顺磁性氧化铁-质粒 DNA（pDNA）负载的微泡（PSp-MBs）与聚焦超声（FUS）和两步磁导航联合使用，以提供空化、质子海绵效应和磁效应，从而提高基因传递效率。

结果

所提出的系统中的基因转染率是商业试剂（TransIT®-LT1）的 2.2 倍。通过空化-磁混合增强细胞膜通透性、质子海绵效应和磁辅助细胞骨架重排，转染率分别提高了约 11%、10%和 6%。体内数据表明，与未经处理的遗传 PD 小鼠相比，该系统的有效基因传递可使多巴胺能神经元的恢复增加 3.2 倍，运动行为改善 3.9 倍。