Six-Month In Vivo Safety Profiling of Topical Ocular AAV5-Decorin Gene Transfer.
Six-Month In Vivo Safety Profiling of Topical Ocular AAV5-Decorin Gene Transfer.
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DOI：
10.1167/tvst.10.10.5
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发表时间：
2021-08-12
影响因子：
3
通讯作者：
Hesemann NP
中科院分区：
文献类型：
--
作者：
Mohan RR;Balne PK;Muayad MS;Tripathi R;Sinha NR;Gupta S;An JA;Sinha PR;Hesemann NP
通过组织选择性局部腺相关病毒 (AAV)5-核心蛋白聚糖 (Dcn) 基因治疗，观察到兔眼体内角膜纤维化和新生血管形成显着缓解。本研究旨在利用兔子模型研究这种基因疗法对眼睛的 6 个月体内毒性分析。在 12 只兔眼中单侧进行小面积上皮刮擦，然后进行角膜干燥，并使用克隆圆筒技术局部递送 AAV5–Dcn (n = 6) 或裸载体 (n = 6)。对侧眼睛作为初始对照（n = 6）。使用多模型临床眼科成像工具（裂隙灯、立体显微镜和 HRT3-RCM 体内共聚焦显微镜）定期对活兔进行安全性和耐受性测量，直到第 6 个月。此后，切除角膜并进行苏木精和伊红染色、梅森毛状体染色、碘化丙啶核染色和定量实时聚合酶链反应分析。基于改良的 Hackett-McDonald 眼部评分系统的临床眼部检查以及角膜体内共聚焦成像显示，在治疗后 6 个月内，接受 AAV5-Dcn 基因转移的兔眼与对照眼相比没有出现眼部毒性迹象（P > 0.05）。组织学和分子分析显示，AAV5-Dcn 与 AAV 裸露或未接触对照组没有显着差异（P > 0.05），并且与未显示异常的隐蔽临床眼科观察结果一致。局部组织靶向的局部 AAV5-Dcn 基因治疗似乎对体内兔眼是安全且无毒的。 AAV5–Dcn 基因疗法有潜力在体内安全地治疗角膜纤维化和新生血管形成，且没有明显的眼毒性。
A significant remission of corneal fibrosis and neovascularization in rabbit eye in vivo was observed from a tissue-selective localized adeno-associated virus (AAV)5–Decorin (Dcn) gene therapy. This study sought to investigate 6-month toxicity profiling of this gene therapy for the eye in vivo using a rabbit model. A small epithelial scrape followed by corneal drying was performed unilaterally in 12 rabbit eyes and either AAV5–Dcn (n = 6) or naked vector (n = 6) was delivered topically using a cloning cylinder technique. Contralateral eyes served as naïve control (n = 6). Safety and tolerability measurements in live rabbits were performed periodically until month 6 using multimodel clinical ophthalmic imaging tools—a slit lamp, stereomicroscope, and HRT3-RCM in vivo confocal microscope. Thereafter, corneas were excised and subjected to hematoxylin and eosin staining, Mason trichome staining, propidium iodide nuclear staining, and quantitative real-time polymerase chain reaction analyses. Clinical eye examinations based on the modified Hackett–McDonald ocular scoring system, and in vivo confocal imaging of the cornea showed no signs of ocular toxicity in rabbit eyes given AAV5–Dcn gene transfer vs control eyes (P > 0.05) through 6 months after treatment. The histologic and molecular analyses showed no significant differences in AAV5–Dcn vs AAV naked or naïve control groups (P > 0.05) and were in accordance with the masked clinical ophthalmic observations showing no abnormalities. Topical tissue-targeted localized AAV5–Dcn gene therapy seems to be safe and nontoxic to the rabbit eye in vivo. AAV5–Dcn gene therapy has the potential to treat corneal fibrosis and neovascularization in vivo safely without significant ocular toxicity.
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影响因子：
3.7
作者：
Mohan RR;Sinha S;Tandon A;Gupta R;Tovey JC;Sharma A
通讯作者：
Sharma A
影响因子：
4.4
作者：
Gupta S;Fink MK;Ghosh A;Tripathi R;Sinha PR;Sharma A;Hesemann NP;Chaurasia SS;Giuliano EA;Mohan RR
通讯作者：
Mohan RR
影响因子：
5.2
作者：
Balne, Praveen K.;Sinha, Nishant R.;Hofmann, Alexandria C.;Martin, Lynn M.;Mohan, Rajiv R.
通讯作者：
Mohan, Rajiv R.
影响因子：
3.4
作者：
Pal-Ghosh, Sonali;Pajoohesh-Ganji, Ahdeah;Tadvalkar, Gauri;Stepp, Mary Ann
通讯作者：
Stepp, Mary Ann
影响因子：
4.4
作者：
Mohan, Rajiv R.;Tandon, Ashish;Tovey, Jonathan C. K.
通讯作者：
Tovey, Jonathan C. K.