Subconjunctival injection of recombinant AAV-angiostatinameliorates alkali burn induced corneal angiogenesis

[摘要] Purpose: To evaluate the effect of subconjunctival injection ofrecombinant adeno-associated virus (rAAV)-angiostatin in alkaliburn-induced corneal angiogenesis.Methods: Adeno-associated viral vector-mediated gene delivery intoextraocular tissue was determined by fluorescent microscopy three weeksafter subconjunctival injection of viral vector expressing greenfluorescent protein (rAAV-GFP). Subconjunctival injection of recombinantadeno-associated viral vector expressing human angiostatin(rAAV-angiostatin) and blank rAAV viral vector (control) was performedin the left eye of male Sprague-Dawley rats (n=6). Alkaline induction ofcorneal neovascularization (NV) was performed three weeks later. CornealNV regression was analyzed 7-14 days after alkali burn with slit lampand digital pictures. Transgenic expression of angiostatin in thecornea, conjunctiva, retina, and muscle insertions was confirmed byreverse-transcriptase polymerase chain reaction (RT-PCR).Results: Transgenic GFP gene expression was detected mainly in themuscle fibers at the extraocular muscle (EOM) insertions aftersubconjunctival injection. The area of corneal neovascularization wassignificantly lower in eyes injected with rAAV-angiostatin (p<0.01)than in eyes injected with the control virus. RT-PCR demonstrated thatthe angiostatin gene expression was highly detectable in muscle fibersand not detectable in the conjunctiva, cornea, and retina.Conclusions: Subconjunctival injection of rAAV-angiostatin reducedalkali burn-induced corneal angiogenesis. We proved the concept thatocular gene therapy by subconjunctival injection ofadenovirus-associated gene transfer of angiogenesis inhibitors can be asimple and safe treatment modality that can achieve therapeutic levelsand long lasting effects in the treatment of corneal NV induced byocular surface disorders.

[学科分类] 生物化学/生物物理