Guo Xiaoshuang, Yuan Zihan, Xu Yang, Zhao Xiaotian, Fang Zhiwei, Yuan Wei-En
Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education, and School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China.
Pharmaceutics. 2019 Apr 8;11(4):171. doi: 10.3390/pharmaceutics11040171.
Peripheral arterial disease (PAD) is often characterized by continued reduction in blood flow supply to limbs. Advanced therapeutic strategies like gene therapy could potentially be applied to limb ischemia therapy. However, developing a gene delivery system with low toxicity and high efficiency remains a great challenge. In this study, a one-pot construction was used to integrate vector synthesis and polyplex fabrication simultaneously in a simple and robust manner. We fabricated an interpenetrating gene delivery network through the physical interaction between low-molecular-weight polyethylenimine (PEI 1.8 kDa) and plasmid DNA (pDNA) and the chemical bonding between PEI and glutaraldehyde (GA), which was named the glutaraldehydelinked-branched PEI (GPEI) polyplex. The final GPEI polyplex system was pH-responsive and biodegradable due to the imine linkage and it could successfully deliver desired vascular endothelial growth factor (VEGF) pDNA. Compared with PEI (25 kDa)/pDNA polyplexes, GPEI polyplexes showed lower cytotoxicity and higher transfection efficiency both in vitro and in vivo. In addition, we demonstrated that GPEI polyplexes could efficiently promote the formation of new capillaries in vivo, which may provide a practicable strategy for clinical hindlimb ischemia therapy in the future.
外周动脉疾病（PAD）通常的特征是肢体的血流供应持续减少。像基因治疗这样的先进治疗策略可能会应用于肢体缺血治疗。然而，开发一种低毒性、高效率的基因递送系统仍然是一个巨大的挑战。在本研究中，采用一锅法以简单且稳健的方式同时整合载体合成和多聚体构建。我们通过低分子量聚乙烯亚胺（PEI 1.8 kDa）与质粒DNA（pDNA）之间的物理相互作用以及PEI与戊二醛（GA）之间的化学键合构建了一个互穿基因递送网络，该网络被命名为戊二醛连接的支化PEI（GPEI）多聚体。由于存在亚胺键，最终的GPEI多聚体系统具有pH响应性且可生物降解，并且它能够成功递送所需的血管内皮生长因子（VEGF）pDNA。与PEI（25 kDa）/pDNA多聚体相比，GPEI多聚体在体外和体内均表现出更低的细胞毒性和更高的转染效率。此外，我们证明GPEI多聚体能够在体内有效促进新毛细血管的形成，这可能为未来临床后肢缺血治疗提供一种可行的策略。
