School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Tianjin 300350, China.
Department of Hepatopancreatobiliary and Splenic Medicine, Affiliated Hospital, Logistics University of People's Armed Police Force, 220 Chenglin Road, Tianjin 300162, China.
Biomater Sci. 2020 Apr 15;8(8):2318-2328. doi: 10.1039/c9bm02000d.
In the past decade, the development of gene carriers has been key in enhancing gene therapy. Gene therapy is associated with not only the delivery process but also gene expression as a prominent role. Herein, for the purpose of achieving a novel breakthrough in gene therapy, we creatively proposed a "strengthened gene expression" idea beyond the range of improving the gene carrier. We constructed three types of gene delivery systems, namely, single-pZNF580 delivery system, single-pVEGF165 delivery system, and dual-gene delivery system. These systems possessed approximate same sizes (∼120 nm) and zeta potentials (∼+20 mV), which indicated negligible differences in their cellular uptake. Interestingly, we found that the gene expression of dual-gene groups significantly increased at the level of both mRNA and protein at least 2 times and 1.5 times as high as single-gene groups, respectively. This "1 + 1 > 2" expression effect benefited from the coordinated expression of the angiogenesis-related genes of ZNF580 and VEGF165. Furthermore, the coordinated effect was also confirmed in HUVEC activities such as an obviously enhanced proliferation and migration of the dual-gene group. Rationally, we further evaluated the effects of coordinated interactions on neovascularization. We observed that the statistic tube number of dual-gene groups was approximately 1.44 times as high as that of single-gene groups. More importantly, this enhanced angiogenesis induced by the coordinated expression was also demonstrated in an in vivo environment. Therefore, we believed that the enhanced gene therapy via the gene expression pathway could provide a creative viewpoint for the design of gene delivery system and therapy.
在过去的十年中，基因载体的发展是增强基因治疗的关键。基因治疗不仅与递送过程有关，而且与基因表达有关，是一个突出的作用。在此，为了在基因治疗方面取得新的突破，我们创造性地提出了一个超越改善基因载体范围的“增强基因表达”的想法。我们构建了三种类型的基因传递系统，即单-pZNF580 传递系统、单-pVEGF165 传递系统和双基因传递系统。这些系统具有相近的大小（约 120nm）和zeta 电位（约+20mV），表明它们的细胞摄取没有明显差异。有趣的是，我们发现双基因组的基因表达在 mRNA 和蛋白质水平上至少分别增加了 2 倍和 1.5 倍，明显高于单基因组。这种“1+1>2”的表达效应得益于 ZNF580 和 VEGF165 的血管生成相关基因的协调表达。此外，在 HUVEC 活性如双基因组的明显增强的增殖和迁移中也证实了这种协调效应。合理地，我们进一步评估了协调相互作用对新生血管形成的影响。我们观察到双基因组的统计管数约为单基因组的 1.44 倍。更重要的是，这种由协调表达引起的增强的血管生成也在体内环境中得到了证明。因此，我们相信，通过基因表达途径增强基因治疗可以为基因传递系统和治疗的设计提供一个创造性的观点。
