Aflibercept Traps Galectin-1, an Angiogenic Factor Associated with Diabetic Retinopathy.
Aflibercept Traps Galectin-1, an Angiogenic Factor Associated with Diabetic Retinopathy.
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DOI：
10.1038/srep17946
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发表时间：
2015-12-09
影响因子：
4.6
通讯作者：
Ishida S
中科院分区：
文献类型：
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作者：
Kanda A;Noda K;Saito W;Ishida S
血管内皮生长因子（VEGF）驱动的血管生成有助于多种疾病，包括癌症和增殖性糖尿病视网膜病变（PDR）。在临床上使用的几种VEGF-A阻滞剂中，afliberept是一种基于VEGFR1/ vegfr2的嵌合诱饵受体，与IgG1的Fc片段融合（即VEGFR1/VEGFR2-Fc）。在这里，我们揭示了一种新的抗血管生成功能，除了它对VEGF家族成员的拮抗作用。免疫沉淀和质谱分析鉴定了半乳糖凝集素-1是一种与阿利伯赛相互作用的蛋白。生物层干涉法显示，与VEGFR1-Fc和VEGFR2-Fc相比，阿非利西普与半乳糖凝集素-1结合的亲和力更高，通过肽：n-糖苷酶f使阿非利西普去糖基化来消除半乳糖凝集素-1 mRNA的表达，在体外缺氧刺激和体内诱导糖尿病等疾病增强视网膜LGALS1/半乳糖凝集素-1 mRNA的表达。半乳糖凝集素-1免疫反应性与VEGFR2共定位在手术切除患有PDR的人眼新血管组织中。与非糖尿病对照组相比，PDR眼玻璃体内半乳糖凝集素-1蛋白水平升高，与VEGF-A水平升高无相关性。术前注射贝伐单抗（一种VEGF-A单克隆抗体）可降低VEGF-A水平，但不能降低半乳糖凝集素-1水平。半乳糖凝集素-1在人视网膜微血管内皮细胞中的应用上调了VEGFR2的磷酸化，这一磷酸化被阿非利西普消除。我们目前的研究结果证明了阿伯西普对半乳糖凝集素-1的中和作用，半乳糖凝集素-1是一种独立于VEGF-A与PDR相关的血管生成因子。
Vascular endothelial growth factor (VEGF)-A-driven angiogenesis contributes to various disorders including cancer and proliferative diabetic retinopathy (PDR). Among several VEGF-A blockers clinically used is aflibercept, a chimeric VEGFR1/VEGFR2-based decoy receptor fused to the Fc fragment of IgG1 (i.e., VEGFR1/VEGFR2-Fc). Here, we revealed a novel anti-angiogenic function for aflibercept beyond its antagonism against VEGF family members. Immunoprecipitation and mass spectrometry analyses identified galectin-1 as an aflibercept-interacting protein. Biolayer interferometry revealed aflibercept binding to galectin-1 with higher affinity than VEGFR1-Fc and VEGFR2-Fc, which was abolished by deglycosylation of aflibercept with peptide:N-glycosidase F. Retinal LGALS1/Galectin-1 mRNA expression was enhanced in vitro by hypoxic stimulation and in vivo by induction of diseases including diabetes. Galectin-1 immunoreactivity co-localized with VEGFR2 in neovascular tissues surgically excised from human eyes with PDR. Compared with non-diabetic controls, intravitreal galectin-1 protein levels were elevated in PDR eyes, showing no correlation with increased VEGF-A levels. Preoperative injection of bevacizumab, a monoclonal antibody to VEGF-A, reduced the VEGF-A, but not galectin-1, levels. Galectin-1 application to human retinal microvascular endothelial cells up-regulated VEGFR2 phosphorylation, which was eliminated by aflibercept. Our present findings demonstrated the neutralizing efficacy of aflibercept against galectin-1, an angiogenic factor associated with PDR independently of VEGF-A.