Liu Zhenzhen, Gong Haijun, Zeng Rui, Liang Xuan, Zhang Li-Ming, Yang Liqun, Lan Yuqing
Institute of Polymer Science, School of Chemistry and Chemical Engineering, Key Laboratory of Designed Synthesis and Application of Polymer Material, Key Laboratory for Polymeric Composite and Functional Materials of Ministry of Education, Sun Yat-sen University, Guangzhou, People's Republic of China.
Department of Ophthalmology, Guangdong Provinci Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, People's Republic of China.
Int J Nanomedicine. 2015 Apr 7;10:2735-49. doi: 10.2147/IJN.S75188. eCollection 2015.
A hyperbranched cationic polysaccharide derivative-mediated small interfering (si)RNA interference strategy was proposed to inhibit nuclear transcription factor-kappa B (NF-κB) activation in human retinal pigment epithelial (hRPE) cells for the gene therapy of diabetic retinopathy. Two hyperbranched cationic polysaccharide derivatives containing the same amount of cationic residues, but with different branching structures and molecular weights, including 3-(dimethylamino)-1-propylamine-conjugated glycogen (DMAPA-Glyp) and amylopectin (DMAPA-Amp) derivatives, were developed for the efficient delivery of NF-κB siRNA into hRPE cells. The DMAPA-Glyp derivative showed lower toxicity against hRPE cells. Furthermore, the DMAPA-Glyp derivative more readily condensed siRNA and then formed the nanoparticles attributed to its higher branching architecture when compared to the DMAPA-Amp derivative. Both DMAPA-Glyp/siRNA and DMAPA-Amp/siRNA nanoparticles were able to protect siRNA from degradation by nuclease in 25% fetal bovine serum. The particle sizes of the DMAPA-Glyp/siRNA nanoparticles (70-120 nm) were smaller than those of the DMAPA-Amp/siRNA nanoparticles (130-180 nm) due to the higher branching architecture and lower molecular weight of the DMAPA-Glyp derivative. In addition, the zeta potentials of the DMAPA-Glyp/siRNA nanoparticles were higher than those of the DMAPA-Glyp/siRNA nanoparticles. As a result, siRNA was much more efficiently transferred into hRPE cells using the DMAPA-Glyp/siRNA nanoparticles rather than the DMAPA-Amp/siRNA nanoparticles. This led to significantly high levels of suppression on the expression levels of NF-κB p65 messenger RNA and protein in the cells transfected with DMAPA-Glyp/siRNA nanoparticles. This work provides a potential approach to promote hyperbranched polysaccharide derivatives as nonviral siRNA vectors for the inhibition of NF-κB activation in hRPE cells.
一种超支化阳离子多糖衍生物介导的小干扰（si）RNA干扰策略被提出来，用于抑制人视网膜色素上皮（hRPE）细胞中核转录因子-κB（NF-κB）的激活，以用于糖尿病视网膜病变的基因治疗。开发了两种含有相同数量阳离子残基，但具有不同分支结构和分子量的超支化阳离子多糖衍生物，包括3-（二甲氨基）-1-丙胺共轭糖原（DMAPA-Glyp）和支链淀粉（DMAPA-Amp）衍生物，用于将NF-κB siRNA高效递送至hRPE细胞中。DMAPA-Glyp衍生物对hRPE细胞显示出较低的毒性。此外，与DMAPA-Amp衍生物相比，DMAPA-Glyp衍生物因其更高的分支结构更容易凝聚siRNA，进而形成纳米颗粒。DMAPA-Glyp/siRNA和DMAPA-Amp/siRNA纳米颗粒都能够保护siRNA在25%胎牛血清中不被核酸酶降解。由于DMAPA-Glyp衍生物具有更高的分支结构和更低的分子量，DMAPA-Glyp/siRNA纳米颗粒的粒径（70-120nm）小于DMAPA-Amp/siRNA纳米颗粒的粒径（130-180nm）。此外，DMAPA-Glyp/siRNA纳米颗粒的ζ电位高于DMAPA-Glyp/siRNA纳米颗粒的ζ电位。结果，使用DMAPA-Glyp/siRNA纳米颗粒比使用DMAPA-Amp/siRNA纳米颗粒能更有效地将siRNA转移到hRPE细胞中。这导致在用DMAPA-Glyp/siRNA纳米颗粒转染的细胞中，NF-κB p65信使RNA和蛋白质的表达水平受到显著高水平的抑制。这项工作提供了一种潜在的方法，以促进超支化多糖衍生物作为非病毒siRNA载体，用于抑制hRPE细胞中NF-κB的激活。
