Antibody-Mediated Inhibition of Tspan12 Ameliorates Vasoproliferative Retinopathy Through Suppression of β-Catenin Signaling

• Published in: Circulation (New York, N.Y.) Vol. 136; no. 2; pp. 180 - 195
• Main Authors: Bucher, Felicitas, Zhang, Ding, Aguilar, Edith, Sakimoto, Susumu, Diaz-Aguilar, Sophia, Rosenfeld, Mauricio, Zha, Zhao, Zhang, Hongkai, Friedlander, Martin, Yea, Kyungmoo
• Format: Journal Article
• Language: English
• Published: United States by the American College of Cardiology Foundation and the American Heart Association, Inc 11.07.2017
• Subjects: Amino Acid Sequence; Animals; Antibodies - genetics; Antibodies - pharmacology; Antibodies - therapeutic use; beta Catenin - antagonists & inhibitors; beta Catenin - metabolism; HEK293 Cells; Human Umbilical Vein Endothelial Cells; Humans; Intravitreal Injections; Mice; Mice, 129 Strain; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Retinal Neovascularization - drug therapy; Retinal Neovascularization - metabolism; Signal Transduction - drug effects; Signal Transduction - physiology; Tetraspanins - antagonists & inhibitors; Tetraspanins - metabolism; β-catenin; antibody; vascular disease; retinopathy; VEGF; angiogenesis; therapy; Tetraspanin12
• ISSN: 0009-7322; 1524-4539
• DOI: 10.1161/CIRCULATIONAHA.116.025604

BACKGROUND: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. METHODS: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. RESULTS: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. CONCLUSIONS: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.