Tawfik Mohamed, Zhang Xiwei, Grigartzik Lisa, Heiduschka Peter, Hintz Werner, Henrich-Noack Petra, van Wachem Berend, Sabel Bernhard A
Institute of Medical Psychology, Otto von Guericke University of Magdeburg, Magdeburg, Germany.
Institute of Process Engineering, Otto von Guericke University of Magdeburg, Magdeburg, Germany.
Neural Regen Res. 2021 Dec;16(12):2534-2541. doi: 10.4103/1673-5374.313068.
Apoptosis, a key mechanism of programmed cell death, is triggered by caspase-3 protein and lowering its levels with gene therapy may rescue cell death after central nervous system damage. We developed a novel, non-viral gene therapy to block caspase-3 gene expression using small interfering RNA (siRNA) delivered by polybutylcyanoacrylate nanoparticles (CaspNPs). In vitro CaspNPs significantly blocked caspase-3 protein expression in C6 cells, and when injected intraocularly in vivo, CaspNPs lowered retinal capsase-3 immunofluorescence by 57.9% in rats with optic nerve crush. Longitudinal, repeated retinal ganglion cell counts using confocal neuroimaging showed that post-traumatic cell loss after intraocular CaspNPs injection was only 36.1% versus 63.4% in lesioned controls. Because non-viral gene therapy with siRNA-nanoparticles can selectively silence caspace-3 gene expression and block apoptosis in post-mitotic neurons, siRNA delivery with nanoparticles may be promising for neuroprotection or restoration of central visual system damage and other neurological disorders. The animal study procedures were approved by the German National Act on the use of experimental animals (Ethic Committee Referat Verbraucherschutz, Veterinärangelegenheiten; Landesverwaltungsamt Sachsen-Anhalt, Halle, Germany, # IMP/G/01-1150/12 and # IMP/G/01-1469/17).
细胞凋亡是程序性细胞死亡的关键机制，由半胱天冬酶 - 3蛋白触发，通过基因治疗降低其水平可能挽救中枢神经系统损伤后的细胞死亡。我们开发了一种新型的非病毒基因疗法，使用聚氰基丙烯酸丁酯纳米颗粒（CaspNPs）递送的小干扰RNA（siRNA）来阻断半胱天冬酶 - 3基因的表达。在体外，CaspNPs显著阻断了C6细胞中半胱天冬酶 - 3蛋白的表达，并且当在体内眼内注射时，CaspNPs使视神经挤压大鼠的视网膜半胱天冬酶 - 3免疫荧光降低了57.9%。使用共聚焦神经成像进行的纵向、重复视网膜神经节细胞计数显示，眼内注射CaspNPs后创伤后细胞损失仅为36.1%，而损伤对照组为63.4%。由于用siRNA - 纳米颗粒进行的非病毒基因治疗可以选择性地沉默有丝分裂后神经元中的半胱天冬酶 - 3基因表达并阻断细胞凋亡，因此用纳米颗粒递送siRNA可能有望用于中枢视觉系统损伤和其他神经系统疾病的神经保护或恢复。动物研究程序已获得德国关于实验动物使用的国家法案批准（德国萨克森 - 安哈尔特州哈雷市消费者保护、兽医事务州行政办公室伦理委员会，# IMP/G/01 - 1150/12和# IMP/G/01 - 1469/17）。
