Disabled-2 is Required for Efficient Hemostasis and Platelet Activation by Thrombin in Mice

Overview

Journal Arterioscler Thromb Vasc Biol

Date 2014 Sep 13

PMID 25212232

Citations 19

Authors

Hui-Ju Tsai

Chien-Ling Huang

Yao-Wen Chang

Ding-Yuan Huang

Chung-Ching Lin

Jonathan A Cooper

Ju-Chien Cheng

Ching-Ping Tseng

Affiliations

Soon will be listed here.

Abstract

Objective: The essential role of platelet activation in hemostasis and thrombotic diseases focuses attention on unveiling the underlying intracellular signals of platelet activation. Disabled-2 (Dab2) has been implicated in platelet aggregation and in the control of clotting responses. However, there is not yet any in vivo study to provide direct evidence for the role of Dab2 in hemostasis and platelet activation.

Approach And Results: Megakaryocyte lineage-restricted Dab2 knockout (Dab2(-/-)) mice were generated to delineate in vivo functions of Dab2 in platelets. Dab2(-/-) mice appeared normal in size with prolonged bleeding time and impaired thrombus formation. Although normal in platelet production and granule biogenesis, Dab2(-/-) platelets elicited a selective defect in platelet aggregation and spreading on fibrinogen in response to low concentrations of thrombin, but not other soluble agonists. Investigation of the role of Dab2 in thrombin signaling revealed that Dab2 has no effect on the expression of thrombin receptors and the outside-in signaling. Dab2(-/-) platelets stimulated by low concentrations of thrombin were normal in Gαq-mediated calcium mobilization and protein kinase C activation, but were defective in Gα₁₂/₁₃-mediated RhoA-ROCKII activation. The attenuated Gα₁₂/₁₃ signaling led to impaired ADP release, Akt-mammalian target of rapamycin and integrin αIIbβ3 activation, fibrinogen binding, and clot retraction. The defective responses of Dab2(-/-) platelets to low concentrations of thrombin stimulation may contribute to the impaired hemostasis and thrombosis of Dab2(-/-) mice.

Conclusions: This study sheds new insight in platelet biology and represents the first report demonstrating that Dab2 is a key regulator of hemostasis and thrombosis by functional interplay with Gα₁₂/₁₃-mediated thrombin signaling.

Citing Articles

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References

Cho M, Pestina T, Steward S, Lowell C, Jackson C, Gartner T . Role of the Src family kinase Lyn in TxA2 production, adenosine diphosphate secretion, Akt phosphorylation, and irreversible aggregation in platelets stimulated with gamma-thrombin. Blood. 2002; 99(7):2442-7. DOI: 10.1182/blood.v99.7.2442. View

Keyel P, Mishra S, Roth R, Heuser J, Watkins S, Traub L . A single common portal for clathrin-mediated endocytosis of distinct cargo governed by cargo-selective adaptors. Mol Biol Cell. 2006; 17(10):4300-17. PMC: 1635374. DOI: 10.1091/mbc.e06-05-0421. View

Huang C, Cheng J, Chen J, Tseng C . Evaluation of the role of Disabled-2 in nerve growth factor-mediated neurite outgrowth and cellular signalling. Cell Signal. 2007; 19(6):1339-47. DOI: 10.1016/j.cellsig.2007.01.019. View

Spudich G, Chibalina M, Au J, Arden S, Buss F, Kendrick-Jones J . Myosin VI targeting to clathrin-coated structures and dimerization is mediated by binding to Disabled-2 and PtdIns(4,5)P2. Nat Cell Biol. 2006; 9(2):176-83. PMC: 2670391. DOI: 10.1038/ncb1531. View

Dubois C, Steiner B, Kieffer N, Reigner S . Thrombin binding to GPIbalpha induces platelet aggregation and fibrin clot retraction supported by resting alphaIIbbeta3 interaction with polymerized fibrin. Thromb Haemost. 2003; 89(5):853-65. View

Huang C, Cheng J, Stern A, Hsieh J, Liao C, Tseng C . Disabled-2 is a novel alphaIIb-integrin-binding protein that negatively regulates platelet-fibrinogen interactions and platelet aggregation. J Cell Sci. 2006; 119(Pt 21):4420-30. DOI: 10.1242/jcs.03195. View

Hosaka K, Takeda T, Iino N, Hosojima M, Sato H, Kaseda R . Megalin and nonmuscle myosin heavy chain IIA interact with the adaptor protein Disabled-2 in proximal tubule cells. Kidney Int. 2009; 75(12):1308-1315. DOI: 10.1038/ki.2009.85. View

Morris S, Cooper J . Disabled-2 colocalizes with the LDLR in clathrin-coated pits and interacts with AP-2. Traffic. 2001; 2(2):111-23. DOI: 10.1034/j.1600-0854.2001.020206.x. View

Varga-Szabo D, Pleines I, Nieswandt B . Cell adhesion mechanisms in platelets. Arterioscler Thromb Vasc Biol. 2008; 28(3):403-12. DOI: 10.1161/ATVBAHA.107.150474. View

10.

Covic L, Misra M, Badar J, Singh C, Kuliopulos A . Pepducin-based intervention of thrombin-receptor signaling and systemic platelet activation. Nat Med. 2002; 8(10):1161-5. DOI: 10.1038/nm760. View

11.

Dubois C, Steiner B, Reigner S . Contribution of PAR-1, PAR-4 and GPIbalpha in intracellular signaling leading to the cleavage of the beta3 cytoplasmic domain during thrombin-induced platelet aggregation. Thromb Haemost. 2004; 91(4):733-42. DOI: 10.1160/TH03-06-0391. View

12.

Offermanns S . Activation of platelet function through G protein-coupled receptors. Circ Res. 2006; 99(12):1293-304. DOI: 10.1161/01.RES.0000251742.71301.16. View

13.

Chong A, Pohlman T, Hampton C, Shimamoto A, Mackman N, Verrier E . Tissue factor and thrombin mediate myocardial ischemia-reperfusion injury. Ann Thorac Surg. 2003; 75(2):S649-55. DOI: 10.1016/s0003-4975(02)04691-x. View

14.

Schonberger T, Ziegler M, Borst O, Konrad I, Nieswandt B, Massberg S . The dimeric platelet collagen receptor GPVI-Fc reduces platelet adhesion to activated endothelium and preserves myocardial function after transient ischemia in mice. Am J Physiol Cell Physiol. 2012; 303(7):C757-66. DOI: 10.1152/ajpcell.00060.2012. View

15.

Tiedt R, Schomber T, Hao-Shen H, Skoda R . Pf4-Cre transgenic mice allow the generation of lineage-restricted gene knockouts for studying megakaryocyte and platelet function in vivo. Blood. 2006; 109(4):1503-6. DOI: 10.1182/blood-2006-04-020362. View

16.

Angiolillo D, Capodanno D, Goto S . Platelet thrombin receptor antagonism and atherothrombosis. Eur Heart J. 2009; 31(1):17-28. PMC: 2800923. DOI: 10.1093/eurheartj/ehp504. View

17.

Huang C, Cheng J, Liao C, Stern A, Hsieh J, Wang C . Disabled-2 is a negative regulator of integrin alpha(IIb)beta(3)-mediated fibrinogen adhesion and cell signaling. J Biol Chem. 2004; 279(40):42279-89. DOI: 10.1074/jbc.M402540200. View

18.

Mishra S, Keyel P, Hawryluk M, Agostinelli N, Watkins S, Traub L . Disabled-2 exhibits the properties of a cargo-selective endocytic clathrin adaptor. EMBO J. 2002; 21(18):4915-26. PMC: 126284. DOI: 10.1093/emboj/cdf487. View

19.

Jin J, Mao Y, Thomas D, Kim S, Daniel J, Kunapuli S . RhoA downstream of G(q) and G(12/13) pathways regulates protease-activated receptor-mediated dense granule release in platelets. Biochem Pharmacol. 2008; 77(5):835-44. PMC: 2745276. DOI: 10.1016/j.bcp.2008.11.017. View

20.

Wolfenson H, Lubelski A, Regev T, Klafter J, Henis Y, Geiger B . A role for the juxtamembrane cytoplasm in the molecular dynamics of focal adhesions. PLoS One. 2009; 4(1):e4304. PMC: 2627934. DOI: 10.1371/journal.pone.0004304. View