The Vimentin Rod Domain Blocks P-Selectin-P-Selectin Glycoprotein Ligand 1 Interactions to Attenuate Leukocyte Adhesion to Inflamed Endothelium

Authors

Fong Wilson Lam
Cameron August Brown
Christian Valladolid
Dabel Cynthia Emebo
Timothy Gerald Palzkill
Miguel Angel Cruz

Publication Date

1-1-2020

Journal

Cell Chemical Biology

DOI

10.1016/j.chembiol.2019.10.013

PMID

31735695

PMCID

MC6980656

PubMedCentral® Posted Date

1-16-2021

PubMedCentral® Full Text Version

Author MSS

Published Open-Access

yes

Keywords

Animals, Cell Adhesion, Disease Models, Animal, Endothelium, Endotoxemia, Female, Healthy Volunteers, Human Umbilical Vein Endothelial Cells, Humans, Interferometry, Leukocytes, Male, Membrane Glycoproteins, Molecular Docking Simulation, P-Selectin, Protein Binding, Protein Domains, Recombinant Proteins, Vimentin

Abstract

Acute inflammation begins with leukocyte P-selectin glycoprotein ligand-1 (PSGL-1) binding to P-selectin on inflamed endothelium and platelets. In pathologic conditions, this process may contribute to secondary organ damage, like sepsis-induced liver injury. Therefore, developing novel therapies to attenuate inflammation may be beneficial. We previously reported that recombinant human vimentin (rhVim) binds P-selectin to block leukocyte adhesion to endothelium and platelets. In this study, we used SPOT-peptide arrays to identify the rod domain as the active region within rhVim that interacts with P-selectin. Indeed, recombinant human rod domain of vimentin (rhRod) binds to P-selectin with high affinity, with in silico modeling suggesting that rhRod binds P-selectin at or near the PSGL-1 binding site. Using bio-layer interferometry, rhRod decreases PSGL-1 binding to immobilized P-selectin, corroborating the in silico data. Under parallel-plate flow, rhRod blocks leukocyte adhesion to fibrin(ogen)-captured platelets, P-selectin/Fc-coated channels, and IL-1β/IL-4-co-stimulated human umbilical vein endothelial cells. Finally, using intravital microscopy in endotoxemic C57Bl/6 mice, rhRod co-localizes with P-selectin in the hepatic sinusoids and decreases neutrophil adhesion to hepatic sinusoids. These data suggest a potential role for rhRod in attenuating inflammation through directly blocking P-selectin-PSGL-1 interactions.

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