Endothelial barrier stabilization by a cyclic tandem peptide targeting VE-cadherin transinteraction in vitro and in vivo.
Inflammatory stimuli result in vascular leakage with potentially life threatening consequences. As a key barrier component, loss of vascular endothelial (VE-) cadherin-mediated adhesion often precedes endothelial breakdown. This study aimed to stabilize VE-cadherin transinteraction and endothelial barrier function using peptides targeting the VE-cadherin adhesive interface. After modelling the transinteracting VE-cadherin structure, an inhibiting single peptide (SP) against a VE-cadherin binding pocket was selected, which specifically blocked VE-cadherin transinteraction as analyzed by single molecule atomic force microscopy (AFM). The tandem peptide (TP) consisting of two SP sequences in tandem was designed to strengthen VE-cadherin adhesion by simultaneously binding and cross-bridging two interacting cadherin molecules. Indeed, in AFM experiments TP specifically rendered VE-cadherin transinteraction resistant against an inhibitory monoclonal antibody. Moreover, TP reduced VE-cadherin lateral mobility and enhanced binding of VE-cadherin-coated microbeads to cultured endothelial cells, but acted independently of the actin cytoskeleton. TP also stabilized endothelial barrier properties against the Ca(2+) ionophore A23187 and the inhibitory antibody. Finally, TP abolished endothelial permeability increase induced by tumour necrosis factor-alpha in microperfused venules in vivo. Stabilization of VE-cadherin adhesion by cross-bridging peptides may therefore be a novel therapeutic approach for the treatment of vascular hyperpermeability.
University of Würzburg, Institute of Anatomy and Cell Biology, Koellikerstr. 6, D-97070 Würzburg, Germany.
Wolfgang-Moritz
WM
Heupel
Athina
A
Efthymiadis
Nicolas
N
Schlegel
Thomas
T
Müller
Yvonne
Y
Baumer
Werner
W
Baumgartner
Detlev
D
Drenckhahn
Jens
J
Waschke
D022005:Adherens Junctions / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D016466:CHO Cells; D015820:Cadherins / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002199:Capillary Permeability / Q000187:drug effects*; D002448:Cell Adhesion / Q000187:drug effects*; D003198:Computer Simulation; D017076:Computer-Aided Design; D006224:Cricetinae; D003412:Cricetulus; D004058:Diffusion; D017097:Electric Impedance; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D036681:Fluorescence Recovery After Photobleaching; D006801:Humans; D007476:Ionophores / Q000494:pharmacology; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D018625:Microscopy, Atomic Force; D008958:Models, Molecular; D010456:Peptides, Cyclic / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014699:Venules / Q000187:drug effects / Q000378:metabolism; D000099152:Cadherin 5
D015703:Antigens, CD; D015820:Cadherins; D000001:Calcimycin; D002118:Calcium; D007476:Ionophores; D008164:Luminescent Proteins; D010456:Peptides, Cyclic; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D000099152:Cadherin 5