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Effects of CD25siRNA gene transfer on high-risk rat corneal graft rejection
Qin Qin1, Yunjie Shi1, Qingqing Zhao1
1Department of Ophthalmology, First Teaching Hospital, Chongqing University of Medical Sciences, Chongqing Eye Bank, Chongqing, China.
Background:
Corneal graft rejection is the major cause of corneal failures. Previous studies have shown that the CD25 monoclonal antibody can inhibit corneal graft rejection during the acute phase of rejection in rat models. In the current study, we evaluated the safety and efficacy of the topical Entranster™ vector in rat corneal applications and further investigated the effects of CD25siRNA gene transfer on high-risk rat corneal graft rejection.
Methods:
Fluorescence detection, clinical assessment, hematoxylin and eosin (HE), terminal deoxynucleotidyl transferase biotin-dUTP nick end labelling (TUNEL) and CD11b assays were used to evaluate the safety and efficacy of CD25siRNA gene transfer in normal SD rat corneas. Orthotopic corneal transplants were performed in alkali burned SD rats. Corneal recipients were divided into four groups that received different treatments. Clinical assessment, western blotting, quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry, HE and transmission electron microscopy (TEM) were performed on all grafts.
Results:
Low toxicity, no immunogenicity and high transfection efficiency were observed in rat corneas treated with the Entranster™ vector. Reduced endothelial cell apoptosis, inflammatory cell infiltration and graft neovascularisation were observed in the CD25siRNA treatment groups. The graft survival curves showed that CD25siRNA treatment significantly prolonged graft survival time, with better graft transparency and less graft oedema. Lower CD25 and higher IL-10 expression were detected in the CD25siRNA treatment groups during the study period, and a higher FOXP3 level was found in the CD25siRNA group than in the CD25siRNA-twice group on days 14 and 21 post-operation.
Conclusions:
The Entranster™ vector is an effective vector for corneal gene therapy. CD25 siRNA gene transfer inhibits corneal graft rejection via upregulation of anti-inflammatory molecule expression.
Insights
Topical CD25 siRNA gene transfer using the Entranster™ vector effectively inhibits corneal graft rejection in rats. This approach reduces inflammation and prolongs graft survival, offering a promising strategy for preventing corneal transplant failure.
Area of Science:
- Ophthalmology
- Gene Therapy
- Immunology
Background:
- Corneal graft rejection is a primary cause of corneal graft failure.
- Previous research indicated CD25 monoclonal antibody efficacy in acute rejection phases.
- This study explores topical Entranster™ vector safety and CD25siRNA gene transfer for high-risk corneal grafts.
Purpose of the Study:
- Evaluate the safety and efficacy of the Entranster™ vector for topical corneal gene delivery.
- Investigate the impact of CD25 siRNA gene transfer on high-risk corneal graft rejection in a rat model.
Main Methods:
- Assessed Entranster™ vector safety and transfection efficiency in normal rat corneas.
- Utilized orthotopic corneal transplants in alkali-burned rats with four treatment groups.
- Employed clinical assessment, western blotting, qRT-PCR, immunohistochemistry, HE, and TEM for graft analysis.
Main Results:
- Entranster™ vector demonstrated low toxicity, no immunogenicity, and high transfection efficiency.
- CD25 siRNA treatment reduced endothelial cell apoptosis, inflammation, and neovascularization.
- Significantly prolonged graft survival, improved transparency, and reduced edema were observed with CD25 siRNA.
Conclusions:
- The Entranster™ vector is a safe and effective tool for corneal gene therapy.
- CD25 siRNA gene transfer successfully inhibits corneal graft rejection.
- Upregulation of anti-inflammatory molecules contributes to the therapeutic effect of CD25 siRNA.
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