From Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, CA (F.B., E.A., S.S., S.D.-A., M.R., Z.Z., H.Z., M.F., K.Y.); and Shanghai Institute for Advanced Immunological Studies, ShanghaiTech University, China (D.Z., K.Y.).
Circulation. 2017 Jul 11;136(2):180-195. doi: 10.1161/CIRCULATIONAHA.116.025604. Epub 2017 Mar 29.
Anti-angiogenic biologicals represent an important concept for the treatment of vasoproliferative diseases. However, the need for continued treatment, the presence of nonresponders, and the risk of long-term side effects limit the success of existing therapeutic agents. Although Tspan12 has been shown to regulate retinal vascular development, nothing is known about its involvement in neovascular disease and its potential as a novel therapeutic target for the treatment of vasoproliferative diseases.
Rodent models of retinal neovascular disease, including the mouse model of oxygen-induced retinopathy and the very low density lipoprotein receptor knockout mouse model were analyzed for Tspan/β-catenin regulation. Screening of a phage display of a human combinatorial antibody (Ab) library was used for the development of a high-affinity Ab against Tspan12. Therapeutic effects of the newly developed Ab on vascular endothelial cells were tested in vitro and in vivo in the oxygen-induced retinopathy and very low density lipoprotein receptor knockout mouse model.
The newly developed anti-Tspan12 Ab exhibited potent inhibitory effects on endothelial cell migration and tube formation. Mechanistic studies confirmed that the Ab inhibited the interaction between Tspan12 and Frizzled-4 and effectively modulates β-catenin levels and target genes in vascular endothelial cells. Tspan12/β-catenin signaling was activated in response to acute and chronic stress in the oxygen-induced retinopathy and very low density lipoprotein receptor mouse model of proliferative retinopathy. Intravitreal application of the Ab showed significant therapeutic effects in both models without inducing negative side effects on retina function. Moreover, combined intravitreal injection of the Ab with a known vascular endothelial growth factor inhibitor, Aflibercept, resulted in significant enhancement of the therapeutic efficacy of each monotherapy. Combination therapy with the Tspan12 blocking antibody can be used to reduce anti-vascular endothelial growth factor doses, thus decreasing the risk of long-term off-target effects.
Tspan12/β-catenin signaling is critical for the progression of vasoproliferative disease. The newly developed anti-Tspan12 antibody has therapeutic effects in vasoproliferative retinopathy and can enhance the potency of existing anti- vascular endothelial growth factor agents.
抗血管生成生物制剂是治疗血管增殖性疾病的一个重要概念。然而，现有治疗药物存在需要持续治疗、存在无应答者以及长期副作用风险等问题，限制了其成功应用。尽管 Tspan12 已被证明可调节视网膜血管发育，但对于其在新生血管疾病中的作用以及作为治疗血管增殖性疾病的新治疗靶点的潜力尚不清楚。
分析了包括氧诱导视网膜病变的小鼠模型和极低密度脂蛋白受体敲除小鼠模型在内的视网膜新生血管疾病的啮齿动物模型中 Tspan/β-连环蛋白的调控作用。利用噬菌体展示的人组合抗体文库进行筛选，开发了针对 Tspan12 的高亲和力抗体。在氧诱导视网膜病变和极低密度脂蛋白受体敲除小鼠模型中，体外和体内检测了新开发的 Ab 对血管内皮细胞的治疗作用。
新开发的抗 Tspan12 Ab 对内皮细胞迁移和管状形成具有强大的抑制作用。机制研究证实，该 Ab 抑制了 Tspan12 与 Frizzled-4 的相互作用，并有效调节了血管内皮细胞中的β-连环蛋白水平和靶基因。在氧诱导视网膜病变和极低密度脂蛋白受体小鼠增生性视网膜病变模型中，Tspan12/β-连环蛋白信号在急性和慢性应激下被激活。Ab 的玻璃体内应用在两种模型中均显示出显著的治疗效果，而对视网膜功能没有产生负面的影响。此外，Ab 与已知的血管内皮生长因子抑制剂 Aflibercept 的玻璃体内联合注射导致每种单药治疗的疗效显著增强。Tspan12 阻断抗体的联合治疗可用于减少抗血管内皮生长因子的剂量，从而降低长期脱靶效应的风险。
Tspan12/β-连环蛋白信号通路对于血管增殖性疾病的进展至关重要。新开发的抗 Tspan12 抗体对血管增殖性视网膜病变具有治疗作用，并可增强现有的抗血管内皮生长因子药物的效力。
