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Quantification of Immunostained Caspase-9 in Retinal Tissue
Published on: July 25, 2022
Gene therapy with caspase-3 small interfering RNA-nanoparticles is neuroprotective after optic nerve damage
Mohamed Tawfik1, Xiwei Zhang2, Lisa Grigartzik1
1Institute of Medical Psychology, Otto von Guericke University of Magdeburg, Magdeburg, Germany.
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).
Insights
Gene therapy using nanoparticles effectively reduced caspase-3 levels, a key protein in cell death. This approach shows promise for protecting neurons and restoring vision after central nervous system damage.
Area of Science:
- Neuroscience
- Biotechnology
- Gene Therapy
Background:
- Apoptosis, or programmed cell death, is mediated by caspase-3 protein.
- Central nervous system damage often leads to neuronal cell death.
- Gene therapy offers a potential strategy to mitigate cell death.
Purpose of the Study:
- To develop and evaluate a novel non-viral gene therapy for blocking caspase-3 gene expression.
- To assess the efficacy of siRNA-nanoparticles in preventing neuronal cell death in vivo.
Main Methods:
- Development of polybutylcyanoacrylate nanoparticles (CaspNPs) for delivering small interfering RNA (siRNA) targeting caspase-3.
- In vitro assessment of CaspNPs' ability to block caspase-3 protein expression in C6 cells.
- In vivo intraocular injection of CaspNPs in rats with optic nerve crush, followed by immunofluorescence and cell counting.
Main Results:
- CaspNPs significantly inhibited caspase-3 protein expression in vitro.
- Intraocular CaspNPs reduced retinal caspase-3 immunofluorescence by 57.9% in rats.
- Post-traumatic retinal ganglion cell loss was significantly lower (36.1%) in CaspNP-treated rats compared to controls (63.4%).
Conclusions:
- Non-viral gene therapy using siRNA-nanoparticles can effectively silence caspase-3 expression and inhibit apoptosis in neurons.
- This siRNA delivery system holds promise for neuroprotection and restoration in central visual system damage.
- Nanoparticle-mediated gene therapy may be a viable therapeutic strategy for various neurological disorders.
