Novel integrin-dependent platelet malfunction in siblings with leukocyte adhesion deficiency-III (LAD-III) caused by a point mutation in FERMT3

Summary

Leukocyte adhesion deficiency-III (LAD-III) also called leukocyte adhesion deficiency-1/variant (LAD1v) is a rare congenital disease caused by defective integrin activation of leukocytes and platelets. Patients with LAD-III present with non-purulent infections and increased bleeding symptoms. We report on a novel integrin-dependent platelet dys-function in two brothers with LAD-III syndrome caused by a homozygous mutation 1717C>T in the FERMT3 gene leading to a premature stop codon R573X in the focal adhesion protein kindlin-3. Stimulation of patients´ platelets with all used agonists resulted in a severely decreased binding of soluble fibrinogen indicating a defect in inside-out activation of the integrin α_IIbβ₃ (GPIIb/IIIa). Patients´ platelets did not respond to the α₂β₁-integrin agonist aggretin-A at all. Our data on granula secretion indicate for the first time that the thrombin receptor PAR-4 but not PAR-1 may be important in integrin-triggered granule secretion in response to thrombin. In contrast, collagen mediated platelet granule secretion was not affected in LAD-III-patients. Thus, integrin-signalling may be not essential in collagen-induced granule secretion. The patients’ peripheral blood mononuclear cells showed a severe loss of adhesion capacity to VCAM-1 and to endothelial cells compared to cells from healthy donors. Rap-1 activation after PMA stimulation could be observed in controls´ but not in patients´ cells. After haematogenesis stem cell transplantation (HSCT) the brothers showed no symptoms of bleeding or immunodeficiency and the integrin-dependent platelet and leukocyte functions normalised.

Keywords

Leukocyte adhesion deficiency - platelets - integrin - kindlin-3

• References

• 1 Kuijpers TW, van Lier RA, Hamann D. et al. Leukocyte adhesion deficiency type 1 (LAD-1)/variant. A novel immunodeficiency syndrome characterized by dys-functional beta2 integrins. J Clin Invest 1997; 100: 1725-1733.
• 2 Harris ES, Shigeoka AO, Li W. et al. A novel syndrome of variant leukocyte adhesion deficiency involving defects in adhesion mediated by beta1 and beta2 inte-grins. Blood 2001; 97: 767-776.
• 3 Etzioni A, Alon R. LAD III versus LAD I variant. Blood 2007; 110: 4129-4130.
• 4 Mor-Cohen R, Rosenberg N, Peretz H. et al. Disulfide bond disruption by a beta 3-Cys549Arg mutation in six Jordanian families with Glanzmann thrombasthenia causes diminished production of constitutively active alpha IIb beta 3. Thromb Haemost 2007; 98: 1257-1265.
• 5 Kuijpers TW, van BR, Kamerbeek N. et al. Natural history and early diagnosis of LAD-1/variant syndrome. Blood 2007; 109: 3529-3537.
• 6 Svensson L, Howarth K, McDowall A. et al. Leukocyte adhesion deficiency-III is caused by mutations in KINDLIN3 affecting integrin activation. Nat Med 2009; 15: 306-312.
• 7 Bergmeier W, Goerge T, Wang HW. et al. Mice lacking the signaling molecule CalDAG-GEFI represent a model for leukocyte adhesion deficiency type III. J Clin Invest 2007; 117: 1699-1707.
• 8 Kinashi T, Aker M, Sokolovsky-Eisenberg M. et al. LAD-III, a leukocyte adhesion deficiency syndrome associated with defective Rap1 activation and impaired stabilization of integrin bonds. Blood 2004; 103: 1033-106.
• 9 Kilic SS, Etzioni A. The clinical spectrum of leukocyte adhesion deficiency (LAD) III due to defective CalDAG-GEF1. J Clin Immunol 2009; 29: 117-122.
• 10 Crittenden JR, Bergmeier W, Zhang Y. et al. CalDAG-GEFI integrates signaling for platelet aggregation and thrombus formation. Nat Med 2004; 10: 982-986.
• 11 Mory A, Feigelson SW, Yarali N. et al. Kindlin-3: a new gene involved in the pathogenesis of LAD-III. Blood 2008; 112: 2591.
• 12 Kuijpers TW, van V d, Weterman MA. et al. LAD-1/variant syndrome is caused by mutations in FERMT3. Blood 2009; 113: 4740-4746.
• 13 Malinin NL, Zhang L, Choi J. et al. A point mutation in KINDLIN3 ablates activation of three integrin subfamilies in humans. Nat Med 2009; 15: 313-318.
• 14 Moser M, Legate KR, Zent R. et al. The tail of integrins, talin, and kindlins. Science 2009; 324: 895-899.
• 15 Plow EF, Qin J, Byzova T. Kindling the flame of integrin activation and function with kindlins. Curr Opin Hematol 2009; 16: 323-328.
• 16 Lahav J, Jurk K, Hess O. et al. Sustained integrin ligation involves extracellular free sulfhydryls and enzymatically catalyzed disulfide exchange. Blood 2002; 100: 2472-2478.
• 17 Kehrel B, Kronenberg A, Rauterberg J. et al. Platelets deficient in glycoprotein IIIb aggregate normally to collagens type I and III but not to collagen type V. Blood 1993; 82: 3364-3370.
• 18 Navdaev A, Clemetson JM, Polgar J. et al. Aggretin, a heterodimeric C-type lectin from Calloselasma rhodostoma (malayan pit viper), stimulates platelets by binding to alpha 2beta 1 integrin and glycoprotein Ib, activating Syk and phospholipase Cgamma 2, but does not involve the glycoprotein VI/Fc receptor gamma chain collagen receptor. J Biol Chem 2001; 276: 20882-20889.
• 19 Polgar J, Clemetson JM, Kehrel BE. et al. Platelet activation and signal transduction by convulxin, a C-type lectin from Crotalus durissus terrificus (tropical rattlesnake) venom via the p62/GPVI collagen receptor. J Biol Chem 1997; 272: 13576-13583.
• 20 Steel CM, Philipson J, Arthur E. et al. Possibility of EB virus preferentially transforming a subpopulation of human B lymphocytes. Nature 1977; 270: 729-731.
• 21 Alon R, Etzioni A. LAD-III, a novel group of leukocyte integrin activation deficiencies. Trends Immunol 2003; 24: 561-566.
• 22 Pasvolsky R, Feigelson SW, Kilic SS. et al. A LAD-III syndrome is associated with defective expression of the Rap-1 activator CalDAG-GEFI in lymphocytes, neutrophils, and platelets. J Exp Med 2007; 204: 1571-1582.
• 23 Jurk K, Kehrel BE. Inherited and Acquired Disorders of Platelet Function. Transfus Med Hemother 2007; 34: 6-19.
• 24 Moser M, Bauer M, Schmid S. et al. Kindlin-3 is required for beta2 integrin-mediated leukocyte adhesion to endothelial cells. Nat Med 2009; 15: 300-305.
• 25 Manevich-Mendelson E, Feigelson SW, Pasvolsky R. et al. Loss of Kindlin-3 in LAD-III eliminates LFA-1 but not VLA-4 adhesiveness developed under shear flow conditions. Blood 2009; 114: 2344-2353.
• 26 Moser M, Nieswandt B, Ussar S. et al. Kindlin-3 is essential for integrin activation and platelet aggregation. Nat Med 2008; 14: 325-330.
• 27 Shattil SJ, Kashiwagi H, Pampori N. Integrin signaling: the platelet paradigm. Blood 1998; 91: 2645-2657.
• 28 Woulfe DS. Platelet G protein-coupled receptors in hemostasis and thrombosis. J Thromb Haemost 2005; 03: 2193-2200.
• 29 Voss B, McLaughlin JN, Holinstat M. et al. PAR1, but not PAR4, activates human platelets through a Gi/o/phosphoinositide-3 kinase signaling axis. Mol Pharmacol 2007; 71: 1399-1406.
• 30 Clemetson KJ, Clemetson JM. Collagen receptors as potential targets for novel anti-platelet agents. Curr Pharm Des 2007; 13: 2673-2683.
• 31 Qasim W, Cavazzana-Calvo M, Davies EG. et al. Allogeneic hematopoietic stem-cell transplantation for leukocyte adhesion deficiency. Pediatrics 2009; 123: 836-840.