McColl-Lockwood Laboratory for Muscular Dystrophy Research, Carolinas Medical Center, 1000 Blythe Blvd., Charlotte, NC, 28231, USA.
J Nanobiotechnology. 2020 Feb 18;18(1):34. doi: 10.1186/s12951-020-0586-8.
The polymer-based drug/gene delivery is promising for the treatment of inherent or acquire disease, because of the polymer's structural flexibility, larger capacity for therapeutic agent, low host immunogenicity and less cost. Antisense therapy is an approach to fighting genetic disorders or infections using antisense oligonucleotides (AOs). Unfortunately, the naked AOs showed low therapeutic efficacy in vivo and in clinical trial due to their poor cellular uptake and fast clearance in bloodstream. In this study, a series of triazine-cored amphiphilic polymers (TAPs) were investigated for their potential to enhance delivery of AOs, 2'-O-methyl phosphorothioate RNA (2'-OMePS) and phosphorodiamidate morpholino oligomer (PMO) both in vitro and in vivo.
TAPs significantly enhanced AO-induced exon-skipping in a GFP reporter-based myoblast and myotube culture system, and observed cytotoxicity of the TAPs were lower than Endoporter, Lipofectamine-2000 or PEI 25K. Application of optimized formulations of TAPs with AO targeted to dystrophin exon 23 demonstrated a significant increase in exon-skipping efficiency in dystrophic mdx mice. The best ones for PMO and 2'-OMePS delivery have reached to 11-, 15-fold compared with the AO only in mdx mice, respectively.
The study of triazine-cored amphiphilic polymers for AO delivery in vitro and in mdx mice indicated that the carrier's performances are related to the molecular size, compositions and hydrophilic-lipophilic balance (HLB) of the polymers, as well as the AO's structure. Improved exon-skipping efficiency of AOs observed in vitro and in mdx mice accompanied with low cytotoxicity demonstrated TAP polymers are potentials as safe and effective delivery carrier for gene/drug delivery.
基于聚合物的药物/基因递药系统在治疗固有或获得性疾病方面具有广阔的应用前景，这是因为聚合物具有结构灵活性、更大的治疗剂载量、较低的宿主免疫原性和较低的成本。反义治疗是一种利用反义寡核苷酸（AOs）治疗遗传疾病或感染的方法。不幸的是，由于裸 AOs 在体内和临床试验中的细胞摄取率低且在血液中清除速度快，其治疗效果不佳。在这项研究中，研究了一系列三嗪核两亲聚合物（TAPs），以评估其增强 AOs（2'-O-甲基硫代磷酸酯 RNA（2'-OMePS）和磷酰胺吗啉寡聚物（PMO））在体外和体内递送的潜力。
TAPs 显著增强了 GFP 报告基因肌母细胞和肌管培养系统中的 AO 诱导外显子跳跃，并且观察到 TAPs 的细胞毒性低于 Endoporter、Lipofectamine-2000 或 PEI 25K。应用针对肌营养不良蛋白外显子 23 的优化 TAPs 与 AO 的制剂在肌营养不良症 mdx 小鼠中显示出显著增加的外显子跳跃效率。对于 PMO 和 2'-OMePS 递送，最佳的 TAPs 制剂与仅用 AO 的 mdx 小鼠相比，分别达到 11 倍和 15 倍。
在体外和 mdx 小鼠中研究三嗪核两亲聚合物对 AO 的递药表明，载体的性能与聚合物的分子大小、组成和亲水-亲脂平衡（HLB）以及 AO 的结构有关。在体外和 mdx 小鼠中观察到 AOs 的外显子跳跃效率提高，同时伴随低细胞毒性，表明 TAP 聚合物具有作为安全有效的基因/药物递药载体的潜力。
