Increased monocyte adhesion by endothelial expression of VCAM-1 missense variation in vitro.
OBJECTIVE
In whole genome and single gene analyses, genetic variation at the vascular cell adhesion molecule-1 (VCAM-1) locus has been associated with inflammatory disease and stroke in sickle cell anaemia. In the current work, we investigated the functional impact of VCAM-1 missense variants and their effect on cell-cell adhesion.


RESULTS
To determine the functional in vitro relevance of five missense VCAM-1 variants (S318F; T384A; G413A; L555V; I716L), we generated wild type and single variant VCAM-1 forms [318F, 384A, 413A, 555V, 716L] in EA.hy926 endothelial cells. Real-time PCR, western blot and ELISA analyses revealed significant differences in mRNA and protein levels for VCAM-1 variants. Monocytic cell lines THP-1 and U937 showed significantly increased adhesion to endothelial cells overexpressing VCAM-1 forms 318F, 555V and 716L compared to those overexpressing wild type VCAM-1 (p < 0.05).


CONCLUSIONS
VCAM-1-dependent cell adhesion to endothelial cells in vitro is significantly increased when expressing VCAM-1 missense mutations 318F, 555V and 716L. The underlying mechanism involves altered VCAM-1 protein levels and function. This observation may be of particular relevance for chronic inflammatory pathophysiologic conditions involving cell-cell adhesion such as atherosclerosis and other proinflammatory conditions.
University Hospital Münster, Institute of Sports Medicine, Molecular Genetics of Cardiovascular Disease, Horstmarer Landweg 39, D-48149 Münster, Germany; University Hospital Münster, Internal Medicine D, Nephrology, Hypertension and Rheumatology, Münster, Germany.
Boris
B
Schmitz
Peter
P
Vischer
Eva
E
Brand
Klaus
K
Schmidt-Petersen
Adelheid
A
Korb-Pap
Katrin
K
Guske
Johanna
J
Nedele
Michael
M
Schelleckes
Jan
J
Hillen
Alois
A
Rötrige
Thomas
T
Simmet
Martin
M
Paul
François
F
Cambien
Stefan-Martin
SM
Brand
D000483:Alleles; D050197:Atherosclerosis / Q000473:pathology; D015153:Blotting, Western; D002448:Cell Adhesion; D042783:Endothelial Cells / Q000166:cytology; D004730:Endothelium, Vascular / Q000473:pathology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007249:Inflammation; D009000:Monocytes / Q000166:cytology*; D009154:Mutation; D020125:Mutation, Missense; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013997:Time Factors; D020298:U937 Cells; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics*
D012333:RNA, Messenger; D019010:Vascular Cell Adhesion Molecule-1
Cardiovascular disease; Functional analyses; Missense variants; Monocyte adhesion; Vascular cell adhesion protein-1