Feasibility of direct vitrectomy-sparing subretinal injection for gene delivery in large animals
Open accessAbstract
PURPOSE: To assess the safety and feasibility of direct vitrectomy-sparing subretinal injection for gene delivery in a large animal model. METHODS: The experimental Liběchov minipigs were used for subretinal delivery of a plasmid DNA vector (pS/MAR-CMV-copGFP) with cytomegalovirus (CMV) promoter, green fluorescent protein (GFP) reporter (copGFP) and a scaffold/matrix attachment region (S/MAR) sequence. The eyes were randomized to subretinal injection of the vector following pars plana vitrectomy (control group) or a direct injection without prior vitrectomy surgery (experimental group). Intra- and post-operative observations up to 30 days after surgery were compared. RESULTS: Six eyes of three mini-pigs underwent surgery for delivery into the subretinal space. Two eyes in the control group were operated with a classical approach (lens-sparing vitrectomy and posterior hyaloid detachment). The other four eyes in the experimental group were injected directly with a subretinal cannula without vitrectomy surgery. No adverse events, such as endophthalmitis, retinal detachment and intraocular pressure elevation were observed post-operatively. The eyes in the experimental group had both shorter surgical time and recovery while achieving the same surgical goal. CONCLUSIONS: This pilot study demonstrates that successful subretinal delivery of gene therapy vectors is achievable using a direct injection without prior vitrectomy surgery.
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The NANO SubRet Gateway Device technique reduced operating time and vector reflux from the retinotomy compared with vitrectomized eyes.
“Notably, the technique also reduced operating time and vector reflux from the retinotomy compared with vitrectomized eyes (Stranak et al., 2024).”
From Gene Therapy-Associated Uveitis (GTAU): Understanding and mitigating the adverse immune response in retinal gene therapy (2025) · cites this paper as Stranak et al., 2024
The Results report shorter surgical times without vitrectomy (10–15 versus 15–30 minutes). Table 2 describes reflux as less common without vitrectomy only if the vector is less viscous than the vitreous; the claim omits that condition. The paper describes a vitrectomy-sparing injection technique but does not identify it as the NANO SubRet Gateway Device technique. (LLM, conf=0.86)
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- Authors
- Stranak, Z., Ardan, T., Nemesh, Y., Toms, M., Toualbi, L., Harbottle, R., Ellederova, Z., Lytvynchuk, L., Petrovski, G., Motlik, J., Moosajee, M., & Kozak, I.
- Affiliations
- Department of Ophthalmology, Charles University, Prague and the Kralovske Vinohrady University Hospital, Prague, Czech Republic
- Czech Academy of Sciences (CZ)
- The Francis Crick Institute (GB)
- German Cancer Research Center (DE)
- Libechov Institute of Animal Physiology and Genetics, Czech Academy of Sciences, Libechov, Czech Republic
- Justus-Liebig-Universität Gießen (DE)
- Oslo University Hospital (NO)
- Moorfields Eye Hospital NHS Foundation Trust (GB)
- Department of Vitreoretinal Surgery and Research and Innovation, Moorfields Eye Hospitals UAE, Abu Dhabi
- Corresponding author
- Kozak, Igor
- ORCID
- Venue
- Current Eye Research
- Publisher
- Informa UK Limited
- Published
- 2024-04-26
- Volume
- 49
- Issue
- 8
- Pages
- 879-887
- ISSN
- 0271-3683 (ISSN-L)
- 1460-2202
- OpenAlex
- W4395675238
- Cited by
- 7
- References
- 37
- Citations by year
- 2026: 2 · 2025: 4 · 2024: 1
- FWCI
- 0.95
- Type
- journal article
- Language
- en
- Open access
- green
- discovery.ucl.ac.uk
- Locations
- Current Eye Research (publishedVersion)
- PubMed (publishedVersion)
- UCL Discovery (University College London) (submittedVersion)
- Figshare (submittedVersion, cc-by)
- The Francis Crick Institute (cc-by)
- Funders
- Johannes Amos Comenius Programme (Research Excellence in Regenerative medicine (CZ.02.01.01/00/22_008/0004562)
- AV21 Strategy (Czech Academy of Sciences) and IAPG institutional support (RVO 67985904)
- Moorfields Eye Charity
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Health Sciences › Medicine › Ophthalmology › Retinal Diseases and Treatments
- Keywords
- subretinal gene delivery, plasmid vectors, GFP reporter, minipigs, pars plana vitrectomy, surgical safety, retinal gene therapy
- MeSH terms
- Animals, Disease Models, Animal, Feasibility Studies, Genetic Vectors, Pilot Projects, Plasmids, Retina, Swine, Swine, Miniature, Vitrectomy, Genetic Therapy, Gene Transfer Techniques, Green Fluorescent Proteins, Injections, Intraocular
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