Institute of Pharmaceutical Technology and Biopharmacy, University of Münster, Corrensstraße 48, 48149 Münster, Germany.
Institute of Pharmaceutical Science, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London, SE1 9NH, UK.
Int J Pharm. 2017 Apr 30;522(1-2):198-209. doi: 10.1016/j.ijpharm.2017.03.006. Epub 2017 Mar 6.
Gene therapy bears great potential for the cure of a multitude of human diseases. Research efforts focussed on the use of viral delivery vectors in the past decades, neglecting non-viral gene therapies of physical or chemical origin due to low transfection efficiency. However, side effects such as activation of oncogenes and inflammatory reactions upon immune cell activation are major obstacles impeding the clinical applicability of viral gene therapy vectors. The aim of this study was the development of a non-viral gene delivery system based on plasmid-loaded human serum albumin nanoparticles, which are biocompatible, biodegradable, and non-toxic in relevant concentrations. The surface of said nanoparticles was modified with different cell penetrating peptides, namely Tat, nona-arginine R9, and the penetratin analogue EB1. We hypothesise that the surface modified nanoparticles can effectively enter HEK 293T cells based on the cell penetrating properties of the different peptides attached. A variety of inhibitors were used targeting distinct uptake pathways in an effort to understand the mechanisms utilized by the various cell penetrating peptides on the surface of the nanoparticles. A significant increase in transfection efficiency compared to free DNA or polyplexes was seen for these novel delivery vectors.
基因治疗在治疗多种人类疾病方面具有巨大的潜力。过去几十年的研究重点集中在使用病毒传递载体上，而忽略了物理或化学起源的非病毒基因疗法，因为其转染效率低。然而，诸如癌基因的激活和免疫细胞激活后的炎症反应等副作用是阻碍病毒基因治疗载体临床应用的主要障碍。本研究的目的是开发一种基于负载质粒的人血清白蛋白纳米颗粒的非病毒基因传递系统，该系统在相关浓度下具有生物相容性、可生物降解和无毒。所述纳米颗粒的表面用不同的细胞穿透肽进行修饰，即 Tat、九聚精氨酸 R9 和 penetratin 类似物 EB1。我们假设基于附着的不同肽的细胞穿透特性，表面修饰的纳米颗粒可以有效地进入 HEK 293T 细胞。使用各种抑制剂针对不同的摄取途径，以努力了解表面纳米颗粒上的各种细胞穿透肽所利用的机制。与游离 DNA 或多聚物相比，这些新型递药载体的转染效率显著提高。
