Efficacious and safe tissue-selective controlled gene therapy approaches for the cornea

Area of Science:

• Ophthalmology
• Gene Therapy
• Molecular Biology

Background:

• Untargeted gene delivery hinders gene therapy success.
• Corneal diseases require targeted therapeutic approaches.
• Keratocytes are key targets for corneal gene therapy.

Purpose of the Study:

• To develop efficient and safe tissue-selective gene therapy for corneal keratocytes in vivo.
• To evaluate gene delivery using adeno-associated virus serotype 5 (AAV5) in normal and diseased rabbit corneas.
• To assess the safety and efficacy of a novel topical vector-delivery technique.

Main Methods:

• Topical application of AAV5 expressing green fluorescent protein (GFP) to rabbit corneas using a hair-dryer based technique.
• Induction of corneal fibrosis and neovascularization in rabbit models.
• Assessment of gene expression, localization, and safety via immunocytochemistry, Western blotting, confocal microscopy, and histological analysis.

Main Results:

• A single 2-minute topical AAV5 application efficiently and selectively transduced keratocytes in normal and diseased rabbit corneas.
• Transgene expression was detected from day 3, peaked at day 7, and persisted for at least 16 weeks.
• No significant toxicity or adverse effects were observed in AAV5-treated corneas compared to controls.

Conclusions:

• Targeted gene therapy approaches using AAV5 are safe and effective for delivering genes into corneal keratocytes in vivo.
• The developed vector-delivery technique shows promise for treating corneal disorders.
• This strategy holds potential for future human corneal disease therapies.