A novel antiplatelet antibody therapy that induces cAMP-dependent endocytosis of the GPVI/Fc receptor gamma-chain complex.
Platelet adhesion to vascular subendothelium, mediated in part by interactions between collagen and glycoprotein VI (GPVI) complexed with Fc receptor gamma-chain, is crucial for thrombus formation. Antiplatelet therapy benefits patients with various thrombotic and ischemic diseases, but the safety and efficacy of existing treatments are limited. Recent data suggest GPVI as a promising target for a novel antiplatelet therapy, for example, GPVI-specific Abs that deplete GPVI from the surface of platelets. Here, we characterized GPVI-specific auto-Abs (YA-Abs) from the first reported patient with ongoing platelet GPVI deficiency caused by the YA-Abs. To obtain experimentally useful human GPVI-specific mAbs with characteristics similar to YA-Abs, we generated human GPVI-specific mouse mAbs and selected 2 representative mAbs, mF1201 and mF1232, whose binding to GPVI was inhibited by YA-Abs. In vitro, mF1201, but not mF1232, induced human platelet activation and GPVI shedding, and mF1232 inhibited collagen-induced human platelet aggregation. Administration of mF1201 and mF1232 to monkeys caused GPVI immunodepletion with and without both significant thrombocytopenia and GPVI shedding, respectively. When a human/mouse chimeric form of mF1232 (cF1232) was labeled with a fluorescent endocytosis probe and administered to monkeys, fluorescence increased in circulating platelets and surface GPVI was lost. Loss of platelet surface GPVI mediated by cF1232 was successfully reproduced in vitro in the presence of a cAMP-elevating agent. Thus, we have characterized cAMP-dependent endocytosis of GPVI mediated by a human GPVI-specific mAb as what we believe to be a novel antiplatelet therapy.
Takayama
Hiroshi
H
Department of Health and Nutrition, School of Human Cultures, The University of Shiga Prefecture, Shiga, Japan. htakayam@shc.usp.ac.jp
Hosaka
Yoshitaka
Y
Nakayama
Kazuyuki
K
Shirakawa
Kamon
K
Naitoh
Katsuki
K
Matsusue
Tomokazu
T
Shinozaki
Mikihiko
M
Honda
Motoyasu
M
Yatagai
Yukiko
Y
Kawahara
Tetsushi
T
Hirose
Jiro
J
Yokoyama
Tooru
T
Kurihara
Michiru
M
Furusako
Shoji
S
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D001323:Autoantibodies / Q000235:genetics / Q000276:immunology; D001792:Blood Platelets / Q000276:immunology*; D000242:Cyclic AMP / Q000378:metabolism*; D004705:Endocytosis / Q000502:physiology*; D005260:Female; D006801:Humans; D008252:Macaca fascicularis; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010973:Platelet Adhesiveness / Q000502:physiology; D010975:Platelet Aggregation Inhibitors / Q000276:immunology / Q000627:therapeutic use; D010980:Platelet Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D017452:Receptors, IgG / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D010975:Platelet Aggregation Inhibitors; D010980:Platelet Membrane Glycoproteins; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins; C098341:platelet membrane glycoprotein VI; D000242:Cyclic AMP