Vector delivery technique affects gene transfer in the cornea in vivo

Area of Science:

• Ophthalmology
• Gene Therapy
• Biomaterials Science

Background:

• Gene therapy holds promise for treating ocular diseases.
• Efficient delivery of therapeutic vectors to the cornea is crucial for successful gene therapy.
• Current delivery methods often involve invasive procedures with potential side effects.

Purpose of the Study:

• To investigate if controlled corneal drying enhances vector absorption in mouse, rabbit, and human corneas.
• To evaluate the effects of corneal drying on gene transfer, corneal structure, and inflammatory responses in vivo.

Main Methods:

• Corneal epithelium was removed, followed by controlled drying using a hair dryer for varying durations.
• Balanced salt solution (BSS) absorption was quantified.
• Adeno-associated virus 8 (AAV8) vector was applied to study gene transfer, with analysis of gene expression, morphology, and inflammation.

Main Results:

• Corneal drying significantly increased BSS absorption in a time-dependent manner.
• 50 seconds of drying led to higher transgene delivery in mouse corneas but with mild-to-moderate morphological changes.
• 30 seconds of drying achieved significant gene transfer without compromising corneal morphology, with minimal inflammatory cell infiltration.

Conclusions:

• Controlled corneal drying is an effective method to increase vector absorption.
• Optimized topical application of AAV serotypes following controlled drying can achieve high, targeted gene expression in the cornea.
• This minimally invasive approach offers a promising strategy for ocular gene therapy with reduced side effects.