Carbon Monoxide-Releasing Molecule-401 Suppresses Polymorphonuclear Leukocyte Migratory Potential by Modulating F-Actin Dynamics

Ken Inoue; Eric K Patterson; Alfredo Capretta; Abdel R Lawendy; Douglas D Fraser; Gediminas Cepinskas

doi:10.1016/j.ajpath.2016.12.025

Carbon Monoxide-Releasing Molecule-401 Suppresses Polymorphonuclear Leukocyte Migratory Potential by Modulating F-Actin Dynamics

Am J Pathol. 2017 May;187(5):1121-1133. doi: 10.1016/j.ajpath.2016.12.025. Epub 2017 Mar 17.

Authors

Ken Inoue¹, Eric K Patterson¹, Alfredo Capretta², Abdel R Lawendy³, Douglas D Fraser⁴, Gediminas Cepinskas⁵

Affiliations

¹ Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada.
² Department of Chemistry, McMaster University, Hamilton, Ontario, Canada.
³ Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada; Department of Medical Biophysics, University of Western Ontario, London, Ontario, Canada.
⁴ Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada; Department of Paediatrics, University of Western Ontario, London, Ontario, Canada.
⁵ Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada; Department of Medical Biophysics, University of Western Ontario, London, Ontario, Canada. Electronic address: gcepinsk@uwo.ca.

PMID: 28320610
DOI: 10.1016/j.ajpath.2016.12.025

Abstract

Carbon monoxide-releasing molecules (CORMs) suppress inflammation by reducing polymorphonuclear leukocyte (PMN) recruitment to the affected organs. We investigated modulation of PMN-endothelial cell adhesive interactions by water-soluble CORM-401 using an experimental model of endotoxemia in vitro. Human umbilical vein endothelial cells (HUVEC) grown on laminar-flow perfusion channels were stimulated with 1 μg/mL lipopolysaccharide for 6 hours and perfused with 100 μmol/L CORM-401 (or inactive compound iCORM-401)-pretreated PMN for 5 minutes in the presence of 1.0 dyn/cm² shear stress.

Huvec: PMN co-cultures were perfused for additional 15 minutes with PMN-free medium containing CORM-401/inactive CORM-401. The experiments were videorecorded (phase-contrast microscopy), and PMN adhesion/migration were assessed off-line. In parallel, CORM-401-dependent modulation of PMN chemotaxis, F-actin expression/distribution, and actin-regulating pathways [eg, p21-activated protein kinases (PAK1/2) and extracellular signal-regulated kinase (ERK)/C-Jun N-terminal kinase (JNK) mitogen-activated protein kinases (MAPK)] were assessed in response to N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation. Pretreating PMN with CORM-401 did not suppress PMN adhesion to HUVEC, but significantly reduced PMN transendothelial migration (P < 0.0001) and fMLP-induced PMN chemotaxis (ie, migration directionality and velocity). These changes were associated with CORM-401-dependent suppression of F-actin levels/cellular distribution and fMLP-induced phosphorylation of PAK1/2 and ERK/JNK MAPK (P < 0.05). CORM-401 had no effect on p38 MAPK activation. In summary, this study demonstrates, for the first time, CORM-401-dependent suppression of neutrophil migratory potential associated with modulation of PAK1/2 and ERK/JNK MAPK signaling and F-actin dynamics.

MeSH terms

Actins / metabolism
CD18 Antigens / metabolism
Carbon Monoxide / metabolism*
Cell Adhesion / physiology
Cell Movement / physiology*
Cells, Cultured
Human Umbilical Vein Endothelial Cells
Humans
MAP Kinase Kinase 4 / metabolism
MAP Kinase Signaling System / physiology
Mitogen-Activated Protein Kinase Kinases / metabolism
Neutrophils / physiology*
Phosphorylation / physiology
Platelet Endothelial Cell Adhesion Molecule-1 / metabolism

Substances

Actins
CD18 Antigens
Platelet Endothelial Cell Adhesion Molecule-1
Carbon Monoxide
MAP Kinase Kinase 4
Mitogen-Activated Protein Kinase Kinases