Diverging signaling events control the pathway of GPVI down-regulation in vivo.
Coronary artery thrombosis is often initiated by platelet activation on collagen-rich subendothelial layers in the disrupted atherosclerotic plaque. The activating platelet collagen receptor glycoprotein VI (GPVI) noncovalently associates with the Fc receptor gamma-chain (FcRgamma), which signals through its immunoreceptor-tyrosine-based activation motif (ITAM) via the adaptor LAT leading to the activation of phospholipase Cgamma2 (PLCgamma2). GPVI is a promising antithrombotic target as anti-GPVI antibodies induce the irreversible loss of the receptor from circulating platelets by yet undefined mechanisms in humans and mice and long-term antithrombotic protection in the latter. However, the treatment is associated with transient but severe thrombocytopenia and reduced platelet reactivity to thrombin questioning its clinical usefulness. Here we show that GPVI down-regulation occurs through 2 distinct pathways, namely ectodomain shedding or internalization/intracellular clearing, and that both processes are abrogated in mice carrying a point mutation in the FcRgamma-associated ITAM. In mice lacking LAT or PLCgamma2, GPVI shedding is abolished, but the receptor is irreversibly down-regulated through internalization/intracellular clearing. This route of GPVI loss is not associated with thrombocytopenia or altered thrombin responses. These results reveal the existence of 2 distinct signaling pathways downstream of the FcRgamma-ITAM and show that it is possible to uncouple GPVI down-regulation from undesired side effects with obvious therapeutic implications.
Rabie
Tamer
T
University of Würzburg, Rudolf Virchow Center, Deutsche Forschungsgemeinschaft Research Center for Experimental Biomedicine, Würzburg, Germany.
Varga-Szabo
David
D
Bender
Markus
M
Pozgaj
Rastislav
R
Lanza
Francois
F
Saito
Takashi
T
Watson
Stephen P
SP
Nieswandt
Bernhard
B
D000818:Animals; D001792:Blood Platelets / Q000166:cytology / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005342:Fibrinolysis; D005434:Flow Cytometry; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D051966:Phospholipase C gamma / Q000097:blood; D015539:Platelet Activation / Q000502:physiology*; D010980:Platelet Membrane Glycoproteins / Q000235:genetics*; D038981:Receptors, Collagen / Q000235:genetics*; D017452:Receptors, IgG / Q000097:blood; D015398:Signal Transduction
D010980:Platelet Membrane Glycoproteins; D038981:Receptors, Collagen; D017452:Receptors, IgG; C098341:platelet membrane glycoprotein VI; D051966:Phospholipase C gamma