Lee Si Hyung, Sim Kyeong Sun, Kim Chan Yun, Park Tae Kwann
Department of Ophthalmology, College of Medicine, Soonchunhyang University, Cheonan 31151, Republic of Korea.
Department of Ophthalmology, Soonchunhyang University Hospital Bucheon, Bucheon 14584, Republic of Korea.
Mol Ther Methods Clin Dev. 2019 Jul 10;14:197-205. doi: 10.1016/j.omtm.2019.06.009. eCollection 2019 Sep 13.
Adeno-associated viruses (AAVs) are the vector of choice for gene therapy in the eye, and self-complementary AAVs (scAAVs), which do not require second-strand DNA synthesis, can be transduced into cells of the trabecular meshwork (TM). The scAAV transduction patterns in the anterior segment of normotensive eyes have been investigated previously, but those in ocular hypertensive (OHT) eyes have not. We assessed the transduction efficiencies of AAV serotypes 2, 5, and 8 in the anterior-segment structures of the eyes of Sprague-Dawley rats with OHT by circumlimbal suturing, followed 3 days later by intracameral injection of scAAV serotype 2 (scAAV2), scAAV5, or scAAV8 packaged with EGFP. The transduction of scAAV2 and scAAV5 in the TM of OHT rats was markedly enhanced after 1 month, and transduction of scAAV5 was more efficient than that of scAAV2; transduction of scAAV8 into the TM did not occur. The transduction of scAAV2, scAAV5, and scAAV8 was enhanced in the ciliary body, iris, and corneal endothelium of the OHT eyes for 3 months. The expression levels of receptors for scAAV2 and scAAV5 were significantly increased in the OHT compared with control eyes. The results demonstrated that scAAV2 and scAAV5 target the ciliary body and TM in OHT eyes, and that the OHT-related changes in anterior-segment structures enhance scAAV transduction.
腺相关病毒（AAV）是眼部基因治疗的首选载体，而无需进行第二链DNA合成的自我互补AAV（scAAV）可被转导至小梁网（TM）细胞中。此前已对正常眼压眼眼前节的scAAV转导模式进行了研究，但尚未研究高眼压（OHT）眼的转导模式。我们通过角膜缘缝线法评估了AAV 2型、5型和8型在患有OHT的Sprague-Dawley大鼠眼前节结构中的转导效率，3天后通过前房内注射包装有增强绿色荧光蛋白（EGFP）的scAAV 2型（scAAV2）、scAAV5或scAAV8。1个月后，scAAV2和scAAV5在OHT大鼠小梁网中的转导显著增强，且scAAV5的转导效率高于scAAV2；scAAV8未转导至小梁网。在3个月内，scAAV2、scAAV5和scAAV8在OHT眼的睫状体、虹膜和角膜内皮中的转导增强。与对照眼相比，OHT眼中scAAV2和scAAV5受体的表达水平显著增加。结果表明，scAAV2和scAAV5靶向OHT眼中的睫状体和小梁网，且眼前节结构中与OHT相关的变化增强了scAAV的转导。
