中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (12): 2534-2541.doi: 10.4103/1673-5374.313068

• 原著:视神经损伤修复保护与再生 • 上一篇    下一篇

非病毒的caspase-3 siRNA纳米粒子基因治疗可保护损伤的视神经

  

  • 出版日期:2021-12-15 发布日期:2021-05-17
  • 基金资助:
    马格德堡医学院和德国学术交流中心（DAAD）提供的Leistungsorientierte Mittelvergabe（LOM）奖学金资助

Gene therapy with caspase-3 small interfering RNA-nanoparticles is neuroprotective after optic nerve damage

Mohamed Tawfik1, #, Xiwei Zhang2, #, Lisa Grigartzik1, Peter Heiduschka3, Werner Hintz2, Petra Henrich-Noack1, 4, Berend van Wachem2, Bernhard A. Sabel1, 5, *   

  1. 1Institute of Medical Psychology, Otto von Guericke University of Magdeburg, Magdeburg, Germany; 2Institute of Process Engineering, Otto von Guericke University of Magdeburg, Magdeburg, Germany; 3Department of Ophthalmology, Münster University Hospital, Münster, Germany; 4Department of Neurology with Institute of Translational Neurology, Münster University Hospital, Münster, Germany; 5Center of Behavioral Brain Sciences (CBBS), Magdeburg, Germany
  • Online:2021-12-15 Published:2021-05-17
  • Contact: Bernhard A. Sabel, PhD, bernhard.sabel@med.ovgu.de.
  • Supported by:
    MT was funded by the Leistungsorientierte Mittelvergabe (LOM) scholarship offered by the medical faculty of Magdeburg and the Deutscher Akademischer Austauschdienst (DAAD).

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摘要：

细胞凋亡是程序性细胞死亡的关键机制，是由caspase-3蛋白触发的，通过基因治疗降低其水平可以减少中枢神经系统损伤后的细胞死亡。实验开发了一种新颖的非病毒基因疗法，通过使用聚氰基丙烯酸丁酯纳米颗粒（CaspNPs）传递小干扰RNA（siRNA）来阻断caspase-3基因表达。在体外，CaspNPs明显阻断了C6细胞中caspase-3蛋白的表达；CaspNPs眼内注射使视神经挤压伤大鼠中使视网膜capsase-3免疫荧光强度降低了57.9%。使用共聚焦神经成像对视网膜神经节细胞进行纵向重复计数，结果显示眼内注射CaspNPs后创伤细胞仅损失36.1%。因为用siRNA纳米粒子进行非病毒基因治疗可以选择性沉默caspace-3基因的表达并阻断有丝分裂后神经元的凋亡，所以用纳米粒子进行siRNA传递可能有望为促进中枢视觉系统损伤和其他神经系统疾病恢复的有效措施。

https://orcid.org/0000-0002-4472-5543 (Bernhard A. Sabel)

Abstract: 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).

Key words: apoptosis, brain, caspase-3, drug delivery, gene therapy, in vivo confocal neuroimaging, nanoparticles, neurodegeneration, neuroprotection, retina, siRNA

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