采用CRISPR/Cas9技术探讨转录因子Islet-1调控VEGFA的表达在糖尿病视网膜病变中作用的研究

胰岛素增强子结合蛋白1（Islet-1）作为转录因子与糖尿病的发生有关，但其在糖尿病视网膜病变中的作用尚不明确。我们的前期研究表明Islet-1参与缺氧诱导的视网膜新生血管形成，同时与糖尿病患者纤维血管膜的形成和糖尿病模型鼠血-视网膜屏障的破坏有关。但上述研究结果的作用途径及机制尚不清楚。为进一步阐明糖尿病视网膜病变形成中Islet-1在其中所起的作用，我们拟采用CRISPR/Cas9技术调控Islet-1的表达，通过db/db鼠和斑马鱼糖尿病模型，及体外高糖培养的视网膜血管内皮细胞、视网膜周细胞、视网膜色素上皮细胞和视网膜Müller细胞，探讨Islet-1在糖尿病血-视网膜屏障破坏、视网膜新生血管形成中所起的作用，并阐明Islet-1在糖尿病视网膜病变形成中对血管内皮细胞生长因子A(VEGFA)表达的调控作用，为完善糖尿病视网膜病变的发病机制及开创新的治疗途径奠定理论基础。

Insulin gene enhancer binding protein 1( Islet-1), as a transcription factor, is associated with the pathogenesis of diabetes. To date, the function of Islet-1 in diabetic retinopathy remains elusive. Our previous study showed that Islet-1 was involved in hypoxia induced retinal neovascularization. Moreover, Islet-1 was found to participate in the formation of diabetic fibrovascular membrane in diabetic patients and destruction of blood retinal barrier in diabetic mice. However, the mechanism underlying aforementioned studies are unclear. To further elucidate the role of Islet-1 in the diabetic retinopathy, we will use CRISPR/Cas9 technique to regulate Islet-1 expression in db/db mice and diabetic zebrafish model in vivo and cultured retinal vascular endothelial cells, retinal pericytes, retinal pigment epithelial cells and retinal Müller cells under high glucose concentration in vitro，which can demonstrate the function of Islet-1 in blood retinal barrier destruction and retinal angiogenesis. Furthermore, it will uncover whether Islet-1 take part in regulating VEGFA expression during the occurrence and development of diabetic retinopathy. The results of this project will help to further understand the pathogenesis of diabetic retinopathy and to provide the novel therapeutic target for diabetic retinopathy.