Trans-Ocular Electric Current In Vivo Enhances AAV-Mediated Retinal Transduction in Large Animal Eye After...

Area of Science:

• Ophthalmology
• Gene Therapy
• Molecular Biology

Background:

• Adeno-associated viral (AAV) vectors are crucial for ocular gene therapy.
• Intravitreal administration is a common delivery route for retinal gene therapy.
• Enhancing AAV transduction efficiency is critical for therapeutic success.

Purpose of the Study:

• To investigate the efficacy of electric-current vector mobility (ECVM) in augmenting retinal transduction of intravitreal AAV vectors.
• To evaluate ECVM's effect on AAV8-CMV-EGFP in rabbit and nonhuman primate (NHP) retinas.
• To explore potential mechanisms underlying ECVM-mediated enhancement of retinal transduction.

Main Methods:

• Intravitreal injection of AAV8-CMV-EGFP followed by immediate application of electric micro-current in rabbits and macaques.
• In vivo fundus imaging to assess retinal GFP expression.
• Retinal immunohistochemistry for cell-specific transduction analysis.
• Quantitative RT-PCR for bFGF expression and immunohistochemistry for GFAP and vimentin.

Main Results:

• ECVM significantly increased AAV transduction efficiency in rabbit retinas, including inner nuclear layer, ganglion cells, and Müller cells.
• A similar trend of improved transduction was observed in ECVM-treated NHP retinas.
• Electric micro-current upregulated bFGF in Müller cells and induced vimentin-associated ILM structural changes in mouse retina.

Conclusions:

• ECVM effectively promotes the transduction efficiency of intravitreal AAV8-CMV-GFP in rabbit and monkey retinas.
• The findings suggest ECVM is a viable strategy to enhance ocular gene delivery.
• This technique holds translational potential for human ocular gene therapy.