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Long-acting FGF21 inhibits retinal vascular leakage in in vivo and in vitro models

Journal article
Authors Y. Tomita
Z. Fu
Z. Wang
B. Cakir
S. S. Cho
W. Britton
Y. Sun
Ann Hellström
S. Talukdar
L. E. H. Smith
Published in International Journal of Molecular Sciences
Volume 21
Issue 4
ISSN 16616596 (ISSN)
Publication year 2020
Published at Institute of Neuroscience and Physiology, Department of Clinical Neuroscience
Language en
Links dx.doi.org/10.3390/ijms21041188
Keywords Claudin-1, Fibroblast growth factor 21 (FGF21), Machine learning analysis, Tight junction, Vascular endothelial growth factor (VEGF), Vascular leakage
Subject categories Clinical Medicine

Abstract

The aim of the current study was to investigate the impact of long-acting fibroblast growth factor 21 (FGF21) on retinal vascular leakage utilizing machine learning and to clarify the mechanism underlying the protection. To assess the effect on retinal vascular leakage, C57BL/6J mice were pre-treated with long-acting FGF21 analog or vehicle (Phosphate Buffered Saline; PBS) intraperitoneally (i.p.) before induction of retinal vascular leakage with intravitreal injection of mouse (m) vascular endothelial growth factor 164 (VEGF164) or PBS control. Five hours after mVEGF164 injection, we retro-orbitally injected Fluorescein isothiocyanate (FITC) -dextran and quantified fluorescence intensity as a readout of vascular leakage, using the Image Analysis Module with a machine learning algorithm. In FGF21- or vehicle-treated primary human retinal microvascular endothelial cells (HRMECs), cell permeability was induced with human (h) VEGF165 and evaluated using FITC-dextran and trans-endothelial electrical resistance (TEER). Western blots for tight junction markers were performed. Retinal vascular leakage in vivo was reduced in the FGF21 versus vehicle- treated mice. In HRMECs in vitro, FGF21 versus vehicle prevented hVEGF-induced increase in cell permeability, identified with FITC-dextran. FGF21 significantly preserved TEER compared to hVEGF. Taken together, FGF21 regulates permeability through tight junctions; in particular, FGF21 increases Claudin-1 protein levels in hVEGF-induced HRMECs. Long-acting FGF21 may help reduce retinal vascular leakage in retinal disorders and machine learning assessment can help to standardize vascular leakage quantification. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.

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