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.

中文翻译：

---

从单基因到双基因递送纳米系统：协同表达对于促进体内新血管形成至关重要

在过去的十年中，基因载体的开发一直是增强基因治疗的关键。基因治疗不仅与传递过程有关，而且与基因表达密切相关。在本文中，为了在基因治疗中取得新的突破，我们创造性地提出了“增强基因表达”的想法，超出了改善基因载体的范围。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。这些系统具有大约相同的大小（〜120 nm）和ζ电势（〜+ 20 mV），这表明它们的细胞摄取差异可忽略不计。有趣的是，我们发现双基因组的基因表达在mRNA和蛋白质水平上均显着增加了至少2倍和1。分别是单基因组的5倍。这种“ 1 + 1> 2”表达效果得益于ZNF580和VEGF165的血管生成相关基因的协同表达。此外，在HUVEC活动中也证实了协同作用，例如双基因组的增殖和迁移明显增强。合理地，我们进一步评估了协同相互作用对新血管形成的影响。我们观察到，双基因组的统计管数约为单基因组的统计管数的1.44倍。更重要的是，由协同表达诱导的这种增强的血管生成也已在 ZNF580和VEGF165的血管生成相关基因的协同表达受益于2”表达效果。此外，在HUVEC活动中也证实了协同作用，例如双基因组的增殖和迁移明显增强。合理地，我们进一步评估了协同相互作用对新血管形成的影响。我们观察到，双基因组的统计管数约为单基因组的统计管数的1.44倍。更重要的是，由协同表达诱导的这种增强的血管生成也已在 ZNF580和VEGF165的血管生成相关基因的协同表达受益于2”表达效果。此外，在HUVEC活动中也证实了协同作用，例如双基因组的增殖和迁移明显增强。合理地，我们进一步评估了协同相互作用对新血管形成的影响。我们观察到，双基因组的统计管数约为单基因组的统计管数的1.44倍。更重要的是，由协同表达诱导的这种增强的血管生成也已在 我们进一步评估了协同相互作用对新血管形成的影响。我们观察到，双基因组的统计管数约为单基因组的统计管数的1.44倍。更重要的是，由协同表达诱导的这种增强的血管生成也已在 我们进一步评估了协同相互作用对新血管形成的影响。我们观察到，双基因组的统计管数约为单基因组的统计管数的1.44倍。更重要的是，由协同表达诱导的这种增强的血管生成也已在体内环境。因此，我们认为通过基因表达途径增强基因治疗可以为基因递送系统和治疗的设计提供创新的观点。