Part I: Minicircle vector technology limits DNA size restrictions on ex vivo gene delivery using nanoparticle vectors: Overcoming a translational barrier in neural stem cell therapy
Part I: Minicircle vector technology limits DNA size restrictions on ex vivo gene delivery using nanoparticle vectors: Overcoming a translational barrier in neural stem cell therapy
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DOI：
10.1016/j.jconrel.2016.06.024
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发表时间：
2016-09-28
影响因子：
10.8
通讯作者：
Chari, Divya M.
中科院分区：
文献类型：
--
作者：
Fernandes, Alinda R.;Chari, Divya M.
基因工程神经干细胞（NSC）移植群体在再生神经学中提供了关键的益处，用于在离体基因治疗中释放治疗性生物分子。神经干细胞是“难以转染”，但适合“磁转染”。尽管这种方法具有很高的临床潜力，但与大尺寸治疗性DNA构建体相关的低瞬时转染是翻译的关键障碍。我们第一次证明了DNA微环（编码必需基因表达组分但缺乏细菌骨架的小DNA载体，从而与常规质粒相比减小了构建体大小）与磁转染一起部署实现了迄今为止报道的最高的安全的非病毒DNA转染水平（高达54%）。微环功能化磁性纳米颗粒（MNP）介导的基因递送也导致了长达四周的持续基因表达。工程化的NSC的所有子细胞类型（神经元、星形胶质细胞和少突胶质细胞）都被转染（与通常仅产生转染的星形胶质细胞的常规质粒相反），为靶向细胞工程化提供了优势。除了增强MNP作为基因递送载体的功能之外，微环技术还提供了安全性/规模扩大方面的关键益处。因此，我们认为这里使用的融合技术的概念验证作为细胞治疗的临床可转化的遗传修饰策略提供了很高的潜力。(C)2016爱思唯尔B.V.保留所有权利。
Genetically engineered neural stem cell (NSC) transplant populations offer key benefits in regenerative neurology, for release of therapeutic biomolecules in ex vivo gene therapy. NSCs are 'hard-to-transfect' but amenable to 'magnetofection'. Despite the high clinical potential of this approach, the low and transient transfection associated with the large size of therapeutic DNA constructs is a critical barrier to translation. We demonstrate for the first time that DNA minicircles (small DNA vectors encoding essential gene expression components but devoid of a bacterial backbone, thereby reducing construct size versus conventional plasmids) deployed with magnetofection achieve the highest, safe non-viral DNA transfection levels (up to 54%) reported so far for primary NSCs. Minicircle-functionalized magnetic nanoparticle (MNP)-mediated gene delivery also resulted in sustained gene expression for up to four weeks. All daughter cell types of engineered NSCs (neurons, astrocytes and oligodendrocytes) were transfected (in contrast to conventional plasmids which usually yield transfected astrocytes only), offering advantages for targeted cell engineering. In addition to enhancing MNP functionality as gene delivery vectors, minicircle technology provides key benefits from safety/scale up perspectives. Therefore, we consider the proof-of-concept of fusion of technologies used here offers high potential as a clinically translatable genetic modification strategy for cell therapy. (C) 2016 Elsevier B.V. All rights reserved.