The P2Y1 Receptor Is Normal in a Patient Presenting a Severe Deficiency of ADP-induced Platelet Aggregation

Catherine Léon; Catherine Vial; Christian Gachet; Philippe Ohlmann; Béatrice Hechler; Jean-Pierre Cazenave; Anna Lecchi; Marco Cattaneo

doi:10.1055/s-0037-1614570

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1999; 81(05): 775-781
DOI: 10.1055/s-0037-1614570

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Schattauer GmbH

The P2Y₁ Receptor Is Normal in a Patient Presenting a Severe Deficiency of ADP-induced Platelet Aggregation

Further Evidence for a Distinct P2 Receptor Responsible for Adenylyl Cyclase Inhibition

Authors

Catherine Léon

²From the INSERM, Etablissement de Transfusion Sanguine de Strasbourg, Strasbourg
Catherine Vial

²From the INSERM, Etablissement de Transfusion Sanguine de Strasbourg, Strasbourg
Christian Gachet

²From the INSERM, Etablissement de Transfusion Sanguine de Strasbourg, Strasbourg
Philippe Ohlmann

²From the INSERM, Etablissement de Transfusion Sanguine de Strasbourg, Strasbourg
Béatrice Hechler
Jean-Pierre Cazenave

²From the INSERM, Etablissement de Transfusion Sanguine de Strasbourg, Strasbourg
Anna Lecchi

¹France; Angelo Bianchi Bonomi Hemophilia and Thrombosis Center, Institute of Internal Medicine, IRCCS Maggiore Hospital, University of Milano, Italy
Marco Cattaneo

¹France; Angelo Bianchi Bonomi Hemophilia and Thrombosis Center, Institute of Internal Medicine, IRCCS Maggiore Hospital, University of Milano, Italy

Abstract

Summary

ADP is a key stimulus inducing platelet shape change and aggregation, a rise in internal calcium and inhibition of adenylyl cyclase. These signaling pathways are thought to be activated by three independent receptors, but to date only the P2Y₁ receptor responsible for calcium mobilization and the ionotropic P2₁ receptor have been identified. We report here the characteristics of the P2Y₁ receptor in a patient presenting a selective deficiency of ADP-induced aggregation. Cloning of the P2Y₁ gene revealed that the patient’s DNA and mRNA were normal. Pharmacological studies showed that the P2Y₁ receptor was expressed and functional in patient’s platelets. Hence, the P2Y₁ receptor is not the cause of the impaired ADP-induced platelet aggregation in this patient. The P2₁ mRNA was also found to be present and normal. These findings add evidence to previous observations suggesting that a third P2 receptor coupled to adenylyl cyclase may be involved in ADP-induced platelet aggregation.

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References

References
1 Macfarlane DE. Agonists and receptors: adenosine diphosphate. In Responses and Metabolism.. Holmsen H. ed 1987; 19-36.
2 Webb TE, Simon J, Krishek BJ, Bateson AN, Smart TG, King BF, Burn-stock G, Barnard EA. Cloning and functional expression of a brain G-protein-coupled ATP receptor.. FEBS Lett 1993; 324: 219-25.
3 Schachter JB, Li Q, Boyer JL, Nicholas RA, Harden TK. Second messenger cascade specificity and pharmacological selectivity of the human P2Y₁purinoceptor.. Br J Pharmacol 1996; 118: 167-73.
4 Léon C, Hechler B, Vial C, Leray C, Cazenave JP, Gachet C. The human P2Y₁receptor is an ADP receptor antagonized by ATP and expressed in platelets and megakaryoblastic cells.. FEBS Lett 1997; 403: 26-30.
5 Hechler B, Vigne P, Léon C, Breittmayer P, Gachet C, Frelin C. ATP derivatives are antagonists of the P2Y₁receptor: similarities to the platelet ADP receptor.. Mol Pharmacol 1998; 53: 727-33.
6 Boyer JL, Romero-Avila T, Schachter JB, Harden TK. Identification of competitive antagonists of the P2Y₁receptor.. Mol Pharmacol 1996; 50: 1323-9.
7 Hechler B, Léon C, Vial C, Vigne P, Frelin C, Cazenave JP, Gachet C. The P2Y₁receptor is necessary for ADP-induced platelet aggregation.. Blood 1998; 92: 152-9.
8 Jin J, Daniel JL, Kunapuli SP. Molecular basis for ADP-induced platelet activation. II. The P2Y₁receptor mediates ADP-induced intracellular calcium mobilization and shape change in platelets.. J Biol Chem 1998; 273: 2030-4.
9 Savi P, Beauverger P, Labouret C, Delfaud M, Salel V, Kaghad M, Herbert JM. Role of P2Y₁purinoceptor in ADP-induced platelet activation.. FEBS Lett 1998; 422: 291-5.
10 Humphries RG, Tomlinson W, Ingall AG, Cage PA, Leff P. FPL 66096LT: a novel, highly potent and selective antagonist at human platelet P2T-receptors.. Br J Pharmacol 1994; 113: 1057-63.
11 Hechler B, Eckly A, Ohlmann P, Cazenave JP, Gachet C. The P2Y₁receptor, necessary but not sufficient to support full ADP-induced platelet aggregation, is not the target of the drug clopidogrel.. Br J Haematol 1998; 103: 858-66.
12 Daniel JL, Dangelmaier C, Lin L, Ashby B, Smith B, Kunapuli S. Molecular basis for ADP-induced platelet activation. I. Evidence for three distinct ADP receptors on human platelets.. J Biol Chem 1998; 273: 2024-9.
13 Fagura MS, Dainty IA, McKay GD, Kirk IP, Humphries RG, Robertson MJ, Dougall IG, Leff P. P2Y₁-receptors in human platelets which are pharmacologically distinct from P2Y_ADP-receptors.. Br J Pharmacol 1998; 124: 157-64.
14 Jin J, Kunapuli SP. Coactivation of two different G protein-coupled receptors is essential for ADP-induced platelet aggregation.. Proc Natl Acad Sci USA 1998; 95: 8070-4.
15 Ohlmann P, Laugwitz K, Nürnberg B, Spicher K, Schultz G, Cazenave JP, Gachet C. The human platelet ADP receptor activates Gi₂proteins.. Biochem J 1995; 312: 775-9.
16 Vial C, Hechler B, Léon C, Cazenave JP, Gachet C. Presence of P2X₁purinoceptors in human platelets and megakaryoblastic cell lines.. Thromb Haemost 1997; 78: 1500-4.
17 Scase TJ, Heath MF, Allen JM, Sage SO, Evans RJ. Identification of a P2X₁purinoceptor expressed on human platelets.. Biochem Biophys Res Comm 1998; 242: 525-8.
18 Sun B, Li K, Okahara K, Kambayashi I J. P2X₁purinoceptor in human platelets. Molecular cloning and functional characterization after heterologous expression.. J Biol Chem 1998; 273: 11544-7.
19 Clifford EE, Parker K, Humphreys BD, Kertezy SB, Dubyak GR. The P2X₁receptor, an adenosine triphosphate-gated cation channel, is expressed in human platelets but not in human blood leukocytes.. Blood 1998; 91: 3172-81.
20 Mahaut-Smith MP, Sage SO, Rink TJ. Rapid ADP-evoked currents in human platelets recorded with the nystatin permeabilized patch technique.. J Biol Chem 1992; 267: 3060-5.
21 MacKenzie A, Mahaut-Smith MP, Sage SO. Activation of receptor-operated cation channels via P2X₁not P2T purinoceptors in human platelets.. J Biol Chem 1996; 271: 2879-81.
22 Cattaneo M, Lecchi A, Randi AM, McGregor JL, Mannuci PM. Identification of a new congenital defect of platelet function characterized by severe impairment of platelet responses to adenosine diphosphate.. Blood 1992; 80: 2787-96.
23 Nurden P, Savi P, Heilmann E, Bihour C, Herbert JM, Maffrand JP, Nurden A. An inherited bleeding disorder linked to a defective interaction between ADP and its receptor on platelets.. J Clin Invest 1995; 95: 1612-22.
24 Gachet C, Cattaneo M, Ohlmann P, Hechler B, Lecchi A, Chevalier J, Cassel D, Manucci PM, Cazenave JP. Purinoceptors on blood platelets: further pharmacological and clinical evidence to suggest the presence of two ADP receptors.. Br J Haematol 1995; 91: 434-44.
25 Cattaneo M, Lombardi R, Zighetti ML, Gachet C, Ohlmann P, Cazenave JP Mannucci PM. Deficiency of [^³³P]2MeSADP binding sites on platelets with secretion defect, normal granule stores and normal thromboxane A2 production.. Thromb Haemost 1997; 77: 986-90.
26 Cazenave JP, Hemmendinger S, Beretz A, Sutter-Bay A, Launay J. L’agrégation plaquettaire: outil d’investigation clinique et d’étude pharmacologique. Méthodologie.. Ann Biol Clin 1983; 41: 167-79.
27 Léon C, Vial C, Cazenave JP, Gachet C. Cloning and sequencing of a human cDNA encoding endothelial P2Y₁purinoceptor.. Gene 1996; 171: 295-7.
28 Sambrook J, Fritsh EF, Maniatis T. Molecular cloning. A laboratory manual.. Cold Spring Harbor Laboratory Press 1989
29 Munson PJ, Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems.. Anal Biochem 1980; 107: 220-39.
30 Breathnach R, Chambon P. Organization and expression of eukaryotic split genes coding for proteins.. Annu Rev Biochem 1981; 50: 349-83.
31 Ayyanathan K, Webbs TE, Sandhu AK, Athwal RS, Barnard EA, Kunapuli SP. Cloning and chromosomal localization of the human P2Y₁purinoceptor.. Biochem Biophys Res Commun 1996; 218: 783-8.
32 Lanza F, Beretz A, Stierlé A, Hanau D, Kubina M, Cazenave JP. Epinephrine potentiates human platelet activation but is not an aggregating agent.. Am J Physiol 1988; 255: 1276-88.
33 Savi P, Pflieger AM, Herber JM. cAMP is not an important messenger for ADP-induced platelet aggregation.. Blood Coag Fibrin 1996; 7: 249-52.
34 Lévy-Toledano S, Maclouf J, Rosa J-P, Gallet C, Vallès G, Nurden P, Nur-den AT. Abnormal tyrosine phosphorylation linked to a defective interaction between ADP and its receptor on platelets.. Thromb Haemost 1998; 80: 463-8.
35 Savi P, Artçanuthurry V, Bornia J, Grelac F, Maclouf J, Levy-Toledano S, Herbert J-M.. Effect of clopidogrel treatment on ADP-induced phosphorylations in rat platelets.. Brit J Haematol 1997; 97:: 185-91.