Claudin-5 Redistribution Induced by Inflammation Leads to Anti-VEGF-Resistant Diabetic Macular Edema.

Abstract

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.

• Authors: Mitsuru Arima, Shintaro Nakao, Muneo Yamaguchi, Hao Feng, Yuya Fujii, Kensuke Shibata, Iori Wada, Yoshihiro Kaizu, Hamid Ahmadieh, Tatsuro Ishibashi, Alan W Stitt, Koh-Hei Sonoda
• Journal: Diabetes (Diabetes) Vol. 69 Issue 5 Pg. 981-999 (05 2020) ISSN: 1939-327X [Electronic] United States
• PMID: 32139595 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2020 by the American Diabetes Association.

Chemical References

• Angiogenesis Inhibitors
• CLDN5 protein, human
• Claudin-5
• Cytokines
• Recombinant Fusion Proteins
• VEGFA protein, human
• Vascular Endothelial Growth Factor A
• aflibercept
• Bevacizumab
• 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
• Receptors, Vascular Endothelial Growth Factor
• Rock1 protein, mouse
• Rock2 protein, mouse
• rho-Associated Kinases
• fasudil
• Ranibizumab

Topics

• 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine (analogs & derivatives, therapeutic use)
• Angiogenesis Inhibitors (therapeutic use)
• Animals
• Bevacizumab (therapeutic use)
• Cell Line
• Claudin-5 (metabolism)
• Cytokines (genetics, metabolism)
• Diabetes Complications
• Diabetes Mellitus, Experimental (chemically induced, complications)
• Endothelial Cells (drug effects)
• Gene Deletion
• Gene Expression Regulation (drug effects, physiology)
• Humans
• Inflammation (metabolism)
• Macular Edema (drug therapy, etiology)
• Mice, Inbred C57BL
• Ranibizumab (therapeutic use)
• Receptors, Vascular Endothelial Growth Factor (therapeutic use)
• Recombinant Fusion Proteins (therapeutic use)
• Retina (pathology)
• Vascular Endothelial Growth Factor A (antagonists & inhibitors)
• rho-Associated Kinases (genetics, metabolism)