Gene therapy knockdown of VEGFR2 in retinal endothelial cells to treat retinopathy.
Gene therapy knockdown of VEGFR2 in retinal endothelial cells to treat retinopathy.
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DOI：
10.1007/s10456-018-9618-5
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发表时间：
2018-11
期刊：
影响因子：
9.8
通讯作者：
Hartnett ME
中科院分区：
文献类型：
--
作者：
Simmons AB;Bretz CA;Wang H;Kunz E;Hajj K;Kennedy C;Yang Z;Suwanmanee T;Kafri T;Hartnett ME
早产儿视网膜病变（ROP）中血管内皮生长因子（VEGF）的抑制引起了早产儿的关注，因为VEGF对视网膜血管发育以及神经元和神经胶质健康至关重要。本研究验证了内皮细胞特异性敲除VEGF受体2（VEGFR 2）或下游STAT 3将抑制VEGF诱导的视网膜病变而不延迟生理性视网膜血管发育的假设。我们开发了一种内皮细胞特异性慢病毒载体，该载体在VE-钙粘蛋白启动子的控制下将shRNA递送至VEGFR 2或STAT 3和绿色荧光蛋白报告基因。在体外和体内验证了慢病毒载体驱动的shRNA的特异性和功效。在高度代表人类ROP的大鼠氧诱导视网膜病变模型中，测定了VEGF 2或STAT 3的内皮细胞敲低对玻璃体内新生血管形成（IVNV）、生理性视网膜血管发育[评估为外周无血管/总视网膜（AVA）面积]、视网膜结构和视网膜功能的影响。通过将慢病毒载体视网膜下注射到出生后第8天的大鼠幼鼠眼中，在视网膜内皮细胞中特异性靶向敲低VEGFR 2或STAT 3有效地抑制了IVNV，并且敲低VEGFR 2还降低了AVA并增加了视网膜厚度而不改变视网膜功能。总之，我们的结果支持特异性敲除视网膜内皮细胞中的VEGFR 2作为治疗视网膜病变的新治疗方法。本文的在线版本（10.1007/s10456-018-9618-5）包含补充材料，可供授权用户使用。
Inhibition of vascular endothelial growth factor (VEGF) in retinopathy of prematurity (ROP) raises concerns for premature infants because VEGF is essential for retinovascular development as well as neuronal and glial health. This study tested the hypothesis that endothelial cell-specific knockdown of VEGF receptor 2 (VEGFR2), or downstream STAT3, would inhibit VEGF-induced retinopathy without delaying physiologic retinal vascular development. We developed an endothelial cell-specific lentiviral vector that delivered shRNAs to VEGFR2 or STAT3 and a green fluorescent protein reporter under control of the VE-cadherin promoter. The specificity and efficacy of the lentiviral vector-driven shRNAs were validated in vitro and in vivo. In the rat oxygen-induced retinopathy model highly representative of human ROP, the effects of endothelial cell knockdown of VEGFR2 or STAT3 were determined on intravitreal neovascularization (IVNV), physiologic retinal vascular development [assessed as area of peripheral avascular/total retina (AVA)], retinal structure, and retinal function. Targeted knockdown of VEGFR2 or STAT3 specifically in retinal endothelial cells by subretinal injection of lentiviral vectors into postnatal day 8 rat pup eyes efficiently inhibited IVNV, and knockdown of VEGFR2 also reduced AVA and increased retinal thickness without altering retinal function. Taken together, our results support specific knockdown of VEGFR2 in retinal endothelial cells as a novel therapeutic method to treat retinopathy. The online version of this article (10.1007/s10456-018-9618-5) contains supplementary material, which is available to authorized users.
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影响因子：
5.3
作者：
STONE, J;ITIN, A;KESHET, E
通讯作者：
KESHET, E
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