Gene Delivery of a Viral Anti-Inflammatory Protein to Combat Ocular Inflammation
Gene Delivery of a Viral Anti-Inflammatory Protein to Combat Ocular Inflammation
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DOI：
10.1089/hum.2014.089
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发表时间：
2015-01-01
影响因子：
4.2
通讯作者：
Lewin, Alfred S.
中科院分区：
文献类型：
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作者：
Ildefonso, Cristhian J.;Jaime, Henrique;Lewin, Alfred S.
视网膜的炎症是眼部疾病如葡萄膜炎、糖尿病性视网膜病和年龄相关性黄斑变性（AMD）的促成因素。来自粘液瘤病毒的M013免疫调节蛋白已显示干扰涉及NLRP 3炎性体和NF-κ B两者的促炎信号传导途径。我们已经开发并表征了腺相关病毒（AAV）载体，其递送M013蛋白（TatM 013）的可分泌和细胞穿透形式。表达的TatM 013蛋白被分泌并阻断单核细胞来源的细胞中的内毒素诱导的白细胞介素（IL）-1 β的分泌和视网膜色素上皮细胞中的反应性雌二醇诱导的IL-1 β的分泌。在内毒素诱导的葡萄膜炎（EIU）小鼠模型中，在玻璃体内注射携带融合至分泌的绿色荧光蛋白（GFP）标签的TatM 013（sGFP-TatM 013）或GFP的基于AAV 2的载体后，评价AAV递送的TatM 013的局部抗炎作用。在活小鼠中通过荧光眼底镜检查证实sGFP-TatM 013转基因的表达。在EIU中，与注射对照AAV-GFP的眼睛相比，注射AAV-sGFP-TatM 013的眼睛中玻璃体中浸润细胞的数量和IL-1 β的浓度显著降低。这些结果表明，当通过AAV递送时，先天免疫应答的病毒衍生抑制剂可以是用于各种眼部炎性疾病的通用疗法。
Inflammation of the retina is a contributing factor in ocular diseases such as uveitis, diabetic retinopathy, and age-related macular degeneration (AMD). The M013 immunomodulatory protein from myxoma virus has been shown to interfere with the proinflammatory signaling pathways involving both the NLRP3 inflammasome and NF-kappa B. We have developed and characterized an adeno-associated viral (AAV) vector that delivers a secretable and cell-penetrating form of the M013 protein (TatM013). The expressed TatM013 protein was secreted and blocked the endotoxin-induced secretion of interleukin (IL)-1 beta in monocyte-derived cells and the reactive aldehyde-induced secretion of IL-1 beta in retinal pigment epithelium cells. The local anti-inflammatory effects of AAV-delivered TatM013 were evaluated in an endotoxin-induced uveitis (EIU) mouse model after intravitreal injection of mice with an AAV2-based vector carrying either TatM013 fused to a secreted green fluorescent protein (GFP) tag (sGFP-TatM013) or GFP. Expression of the sGFP-TatM013 transgene was demonstrated by fluorescence funduscopy in living mice. In EIU, the number of infiltrating cells and the concentration of IL-1 beta in the vitreous body were significantly lower in the eyes injected with AAV-sGFP-TatM013 compared with the eyes injected with control AAV-GFP. These results suggest that a virus-derived inhibitor of the innate immune response, when delivered via AAV, could be a generalized therapy for various inflammatory diseases of the eye.