title: Facilitated diffusion of VEGF165 through descemet's membrane with sucrose octasulfate.
abstract: Vascular endothelial growth factor A (VEGF-A) is a promoter of neovascularization and thus a popular therapeutic target for diseases involving excessive growth of blood vessels. In this study, we explored the potential of the disaccharide sucrose octasulfate (SOS) to alter VEGF165 diffusion through Descemet's membrane. Descemet's membranes were isolated from bovine eyes and used as a barrier between two chambers of a diffusion apparatus to measure VEGF transport. Diffusion studies revealed a dramatic increase in VEGF165 transport in the presence of SOS, with little diffusion of VEGF165 across the membrane over a 10-h time course in the absence of SOS. Diffusion studies with VEGF121, a non-heparin binding variant of VEGF, showed robust diffusion with or without SOS. To determine a possible mechanism, we measured the ability of SOS to inhibit VEGF interactions with extracellular matrix (ECM), using cell-free and cell surface binding assays. Binding studies showed SOS had no effect on VEGF165 binding to either heparin-coated plates or endothelial cell surfaces at less than mg/ml concentrations. In contrast, we show that SOS inhibited VEGF165 binding to fibronectin in a dose dependent manner and dramatically accelerated the rate of release of VEGF165 from fibronectin. SOS also inhibited the binding of VEGF165 to fibronectin-rich ECM deposited by vascular smooth muscle cells. These results suggest that fibronectin-rich extracellular matrices serve as barriers to VEGF165 diffusion by providing a network of binding sites that can trap and sequester the protein. Since the content of Descemet's membrane is typical of many basement membranes it is possible that they serve throughout the body as formidable barriers to VEGF165 diffusion and tightly regulate its bioavailability and distribution within tissues.
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Kentucky, Lexington, KY 40536-0305, USA. michael.fannon@uky.edu
authors.forename: Michael
authors.initials: M
authors.lastname: Fannon
authors.forename: Kimberly
authors.initials: K
authors.lastname: Forsten-Williams
authors.forename: Bing
authors.initials: B
authors.lastname: Zhao
authors.forename: Eva
authors.initials: E
authors.lastname: Bach
authors.forename: Parag P
authors.initials: PP
authors.lastname: Parekh
authors.forename: Chia Lin
authors.initials: CL
authors.lastname: Chu
authors.forename: Adrienne L
authors.initials: AL
authors.lastname: Goerges-Wildt
authors.forename: Jo Ann
authors.initials: JA
authors.lastname: Buczek-Thomas
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Nugent
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D003886:Descemet Membrane / Q000187:drug effects / Q000378:metabolism; D015194:Diffusion Chambers, Culture; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism; D055459:Facilitated Diffusion / Q000187:drug effects*; D005353:Fibronectins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D013395:Sucrose / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry / Q000378:metabolism*
chemical_list: D005353:Fibronectins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D013395:Sucrose; C053081:sucrose octasulfate