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Updated: May 10, 2026

Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium
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Retinal gene therapy using epiretinal AAV-containing fibrin hydrogel implants.

Brittni A Scruggs1,2, Aubrey Berger1,3, Travis Knudsen1

  • 1Department of Ophthalmology, Mayo Clinic, Rochester, MN, USA.

Science Advances
|September 5, 2025
PubMed
Summary

Epiretinal hydrogels carrying adeno-associated virus (AAV) gene therapy offer a safer, more effective alternative to subretinal injections. This novel approach broadens retinal pigment epithelium transduction without inflammation or atrophy.

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Area of Science:

  • Ophthalmology
  • Gene Therapy
  • Biomaterials

Background:

  • Subretinal injection of adeno-associated virus (AAV) is a common route for retinal gene therapy but can cause adverse events like chorioretinal atrophy.
  • Current subretinal delivery methods often result in localized transduction, limiting therapeutic efficacy.
  • Intravitreal injections, while less invasive, typically lead to insufficient transduction and inflammation.

Purpose of the Study:

  • To develop and evaluate a novel epiretinal hydrogel delivery system for AAV gene therapy.
  • To compare the efficacy and safety of epiretinal hydrogel delivery with traditional subretinal and intravitreal injection methods.
  • To assess the potential of this new technology for treating retinal degenerations.

Main Methods:

  • High-concentration fibrin hydrogels were manufactured to encapsulate adeno-associated virus serotype 2 carrying the green fluorescent protein (AAV2-GFP).
  • The hydrogels were characterized for AAV distribution, mechanical properties, and infectivity.
  • Epiretinal, subretinal, and intravitreal placements of AAV2-GFP were performed in animal models (n=11, n=5, and n=3, respectively).

Main Results:

  • Subretinal injection resulted in inconsistent retinal pigment epithelium (RPE) transduction, localized delivery, and severe atrophy in two cases.
  • Intravitreal injection showed weak transduction and induced inflammation.
  • Epiretinal hydrogels degraded within days, achieving broad RPE transduction without observable atrophy or inflammation.

Conclusions:

  • Epiretinal placement of fibrin-AAV hydrogels provides a promising alternative for retinal gene therapy delivery.
  • This method overcomes the limitations of subretinal and intravitreal injections, offering improved safety and efficacy.
  • The technology holds potential for advancing treatments for various ocular and systemic diseases.