Biodistribution of adeno-associated virus type 2 carrying multi-characteristic opsin in dogs following intravitreal injection.
Biodistribution of adeno-associated virus type 2 carrying multi-characteristic opsin in dogs following intravitreal injection.
复制标题
玻璃体内注射后，与腺相关的2型腺相关病毒的生物分布。
DOI：
10.1111/jcmm.16823
复制
发表时间：
2021-09
影响因子：
5.3
通讯作者：
Mohanty SK
中科院分区：
文献类型：
--
作者：
Tchedre KT;Batabyal S;Galicia M;Narcisse D;Mustafi SM;Ayyagari A;Chavala S;Mohanty SK
基于重组腺相关病毒(RAAV)载体的视网膜疾病的基因治疗已经取得了临床成功，基于rAAV的光遗传疗法的临床试验目前正在进行中。最近，我们开发了多特征视蛋白(MCO)，它被证明能有效地重新光敏小鼠的光感受器退化的视网膜，导致在环境光环境下的视力恢复。在这里，我们报告了携带rAAV2的MCO(vMCO-I)在野生型犬眼内分娩后在活体样本和死后器官中的生物分布。免疫组织化学结果显示，玻璃体内注射vMCO-I可导致内核层(INL)的基因转导，但不会引起明显的炎症或免疫反应。应用定量聚合酶链式反应(QPCR)对活体排泄物和唾液、鼻腔分泌物等体液进行载体DNA分析，发现鼻腔分泌物或唾液中的媒介拷贝没有相关性增加，低剂量组在注射后13周尿液中媒介拷贝的增加幅度很小，高剂量组在注射后3~13周的粪便中媒介拷贝的增加幅度很小，提示病毒载体通过尿液和粪便清除。进一步对从粪便中提取的载体DNA进行PCR分析，发现注射后3周没有转基因。玻璃体内注射vMCO-I后，在肠系膜淋巴结、肝脏、脾和睾丸中很少出现零星的非靶向表达。这项研究表明，基于rAAV2的MCO-I玻璃体内注射用于视网膜基因治疗是安全的。
Gene therapy of retinal diseases using recombinant adeno‐associated virus (rAAV) vector‐based delivery has shown clinical success, and clinical trials based on rAAV‐based optogenetic therapies are currently in progress. Recently, we have developed multi‐characteristic opsin (MCO), which has been shown to effectively re‐photosensitize photoreceptor‐degenerated retina in mice leading to vision restoration at ambient light environment. Here, we report the biodistribution of the rAAV2 carried MCO (vMCO‐I) in live samples and post‐mortem organs following intraocular delivery in wild‐type dogs. Immunohistochemistry showed that the intravitreal injection of vMCO‐I resulted in gene transduction in the inner nuclear layer (INL) but did not induce detectable inflammatory or immune reaction in the dog retina. Vector DNA analysis of live body wastes and body fluids such as saliva and nasal secretions using quantitative polymerase chain reaction (qPCR) showed no correlative increase of vector copy in nasal secretions or saliva, minimal increase of vector copy in urine in the low‐dose group 13 weeks after injection and in the faeces of the high‐dose group at 3–13 weeks after injection suggesting clearance of the virus vector via urine and faeces. Further analysis of vector DNA extracted from faeces using PCR showed no transgene after 3 weeks post‐injection. Intravitreal injection of vMCO‐I resulted in few sporadic off‐target presences of the vector in the mesenteric lymph node, liver, spleen and testis. This study showed that intravitreal rAAV2‐based delivery of MCO‐I for retinal gene therapy is safe.
登录
查看更多内容
影响因子：
4.2
作者：
Kattenhorn LM;Tipper CH;Stoica L;Geraghty DS;Wright TL;Clark KR;Wadsworth SC
通讯作者：
Wadsworth SC
影响因子：
12.4
作者：
Cearley, CN;Wolfe, JH
通讯作者：
Wolfe, JH
影响因子：
3.5
作者：
Fitzsimons, Helen L.;Riban, Veronique;Bland, Ross J.;Wendelken, Jennifer L.;Sapan, Christine V.;During, Matthew J.
通讯作者：
During, Matthew J.
影响因子：
5.1
作者：
Hickey DG;Edwards TL;Barnard AR;Singh MS;de Silva SR;McClements ME;Flannery JG;Hankins MW;MacLaren RE
通讯作者：
MacLaren RE
影响因子：
--
作者：
Ai, Jianzhong;Ibraheim, Raed;Gao, Guangping
通讯作者：
Gao, Guangping