Valdés-Sánchez Lourdes, Borrego-González Sara, Montero-Sánchez Adoración, Massalini Simone, de la Cerda Berta, Díaz-Cuenca Aránzazu, Díaz-Corrales Francisco J
Regeneration and Cell Therapy Department, Andalusian Molecular Biology and Regenerative Medicine Centre (CABIMER), Junta de Andalucía, CSIC, Universidad de Sevilla, Universidad Pablo de Olavide, 41092 Sevilla, Spain.
Materials Science Institute of Seville (ICMS), Joint CSIC-University of Seville Center, 41092 Seville, Spain.
J Clin Med. 2022 Apr 13;11(8):2170. doi: 10.3390/jcm11082170.
Gene therapy is a therapeutic possibility for retinitis pigmentosa (RP), in which therapeutic transgenes are currently delivered to the retina by adeno-associated viral vectors (AAVs). Although their safety and efficacy have been demonstrated in both clinical and preclinical settings, AAVs present some technical handicaps, such as limited cargo capacity and possible immunogenicity in repetitive doses. The development of alternative, non-viral delivery platforms like nanoparticles is of great interest to extend the application of gene therapy for RP.
Amino-functionalized mesoporous silica-based nanoparticles (N-MSiNPs) were synthesized, physico-chemically characterized, and evaluated as gene delivery systems for human cells in vitro and for retinal cells in vivo. Transgene expression was evaluated by WB and immunofluorescence. The safety evaluation of mice subjected to subretinal injection was assessed by ophthalmological tests (electroretinogram, funduscopy, tomography, and optokinetic test).
N-MSiNPs delivered transgenes to human cells in vitro and to retinal cells in vivo. No adverse effects were detected for the integrity of the retinal tissue or the visual function of treated eyes. N-MSiNPs were able to deliver a therapeutic transgene candidate for RP, , both in vitro and in vivo.
N-MSiNPs are safe for retinal delivery and thus a potential alternative to viral vectors.
基因治疗是视网膜色素变性（RP）的一种治疗可能性，目前治疗性转基因是通过腺相关病毒载体（AAV）递送至视网膜的。尽管其安全性和有效性已在临床和临床前环境中得到证实，但AAV存在一些技术障碍，如有限的载量和重复给药时可能的免疫原性。开发像纳米颗粒这样的替代性非病毒递送平台对于扩展基因治疗在RP中的应用具有重大意义。
合成了氨基功能化的介孔二氧化硅基纳米颗粒（N-MSiNPs），对其进行了物理化学表征，并在体外对人类细胞以及在体内对视网膜细胞作为基因递送系统进行了评估。通过蛋白质免疫印迹法（WB）和免疫荧光评估转基因表达。通过眼科检查（视网膜电图、眼底镜检查、断层扫描和视动试验）对接受视网膜下注射的小鼠进行安全性评估。
N-MSiNPs在体外将转基因递送至人类细胞，并在体内递送至视网膜细胞。未检测到对视网膜组织完整性或治疗后眼睛视觉功能的不良影响。N-MSiNPs能够在体外和体内递送一种用于RP的治疗性转基因候选物。
N-MSiNPs用于视网膜递送是安全的，因此是病毒载体的一种潜在替代品。
