Fluorinated per-acetylated GalNAc metabolically alters glycan structures on leukocyte PSGL-1 and reduces cell binding to selectins.
Novel strategies to control the binding of adhesion molecules belonging to the selectin family are required for the treatment of inflammatory diseases. We tested the possibility that synthetic monosaccharide analogs can compete with naturally occurring sugars to alter the O-glycan content on human leukocyte cell surface selectin-ligand, P-selectin glycoprotein ligand-1 (PSGL-1). Resulting reduction in the sialyl Lewis-X-bearing epitopes on this ligand may reduce cell adhesion. Consistent with this hypothesis, 50muM per-acetylated 4F-GalNAc added to the growth media of promyelocytic HL-60 cells reduced the expression of the cutaneous lymphocyte associated-antigen (HECA-452 epitope) by 82% within 2 cell doubling cycles. Cell binding to all 3 selectins (L-, E-, and P-selectin) was reduced in vitro. 4F-GalNAc was metabolically incorporated into PSGL-1, and this was accompanied by an approximately 20% reduction in PSGL-1 glycan content. A 70% to 85% reduction in HECA-452 binding epitope and N-acetyl lactosamine content in PSGL-1 was also noted on 4F-GalNAc addition. Intravenous 4F-GalNAc infusion reduced leukocyte migration to the peritoneum in a murine model of thioglycolate-induced peritonitis. Thus, the compound has pharmacologic activity. Overall, the data suggest that 4F-GalNAc may be applied as a metabolic inhibitor to reduce O-linked glycosylation, sialyl Lewis-X formation, and leukocyte adhesion via the selectins.
Marathe
Dhananjay D
DD
Chemical and Biological Engineering, State University of New York at Buffalo, Buffalo, NY, USA.
Buffone
Alexander
A
Chandrasekaran
E V
EV
Xue
Jun
J
Locke
Robert D
RD
Nasirikenari
Mehrab
M
Lau
Joseph T Y
JT
Matta
Khushi L
KL
Neelamegham
Sriram
S
D000107:Acetylation; D000117:Acetylglucosamine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000378:metabolism; D002448:Cell Adhesion; D002465:Cell Movement; D002634:Chemotaxis, Leukocyte; D004195:Disease Models, Animal; D005434:Flow Cytometry; D006031:Glycosylation; D018922:HL-60 Cells; D006801:Humans; D007962:Leukocytes / Q000378:metabolism*; D007983:Lewis Blood Group Antigens / Q000276:immunology / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009363:Neoplasm Proteins / Q000276:immunology / Q000378:metabolism; D010538:Peritonitis / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D011134:Polysaccharides / Q000737:chemistry*; D011485:Protein Binding
C445125:HECA protein, human; D007983:Lewis Blood Group Antigens; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; C084882:P-selectin ligand protein; D011134:Polysaccharides; C089178:2-acetamido-1,3,6-tri-O-acetyl-4-deoxy-4-fluoroglucopyranose; D000117:Acetylglucosamine