Claudin-5 Redistribution Induced by Inflammation Leads to Anti-VEGF-Resistant Diabetic Macular Edema
Claudin-5 Redistribution Induced by Inflammation Leads to Anti-VEGF-Resistant Diabetic Macular Edema
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DOI：
10.2337/db19-1121
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发表时间：
2020-05-01
期刊：
影响因子：
7.7
通讯作者：
Sonoda, Koh-Hei
中科院分区：
文献类型：
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作者：
Arima, Mitsuru;Nakao, Shintaro;Sonoda, Koh-Hei
大约40%的糖尿病性黄斑水肿（DME）患者对抗血管内皮生长因子（VEGF）治疗（rDME）有抵抗力。在这里，我们证明了炎性细胞因子和VEGF之间的显着相关性，如在幼稚DME中观察到的，在rDME患者中丢失。VEGF在小鼠视网膜中的过度表达引起延迟的炎性细胞因子上调、单核细胞/巨噬细胞浸润（CD 11b（+）Ly 6C（+）CCR 2（+）细胞）、巨噬细胞/小胶质细胞活化（CD 11b（+）CD 80（+）细胞）和由于claudin-5重新分布引起的血视网膜屏障破坏，其不能通过单独的VEGF阻断而恢复。VEGF过度表达的视网膜的磷酸化蛋白质分析显示rho相关的卷曲螺旋蛋白激酶（ROCK）激活。给予选择性ROCK抑制剂Ripasudil，可减轻视网膜炎症和claudin-5再分布。Ripasudil还通过在炎症性血管紧张素刺激的内皮中的转录增强和降解抑制来促进claudin-5表达的稳定性。值得注意的是，抗VEGF剂和ROCK抑制剂在抑制细胞因子上调、单核细胞/巨噬细胞浸润、巨噬细胞/小胶质细胞活化和密蛋白-5再分布方面具有协同作用。此外，体外分析证实，claudin-5再分布依赖于ROCK 2，而不是ROCK 1。这种协同效应也在人rDME病例中得到证实。我们的研究结果表明，ROCK介导的claudin-5的炎症再分布是DME抗VEGF抵抗的关键机制。
Approximately 40% of patients with diabetic macular edema (DME) are resistant to anti-vascular endothelial growth factor (VEGF) therapy (rDME). Here, we demonstrate that significant correlations between inflammatory cytokines and VEGF, as observed in naive DME, are lost in patients with rDME. VEGF overexpression in the mouse retina caused delayed inflammatory cytokine upregulation, monocyte/macrophage infiltration (CD11b(+) Ly6C(+) CCR2(+) cells), macrophage/microglia activation (CD11b(+) CD80(+) cells), and blood-retinal barrier disruption due to claudin-5 redistribution, which did not recover with VEGF blockade alone. Phosphorylated protein analysis of VEGF-overexpressed retinas revealed rho-associated coiled-coil-containing protein kinase (ROCK) activation. Administration of ripasudil, a selective ROCK inhibitor, attenuated retinal inflammation and claudin-5 redistribution. Ripasudil also contributed to the stability of claudin-5 expression by both transcriptional enhancement and degradation suppression in inflammatory cytokine-stimulated endothelium. Notably, the anti-VEGF agent and the ROCK inhibitor were synergic in suppressing cytokine upregulation, monocyte/macrophage infiltration, macrophage/microglia activation, and claudin-5 redistribution. Furthermore, in vitro analysis confirmed that claudin-5 redistribution depends on ROCK2 but not on ROCK1. This synergistic effect was also confirmed in human rDME cases. Our results suggest that ROCK-mediated claudin-5 redistribution by inflammation is a key mechanism in the anti-VEGF resistance of DME.