Filamin-A is required for the incorporation of tissue factor into cell-derived microvesicles

Summary

We previously reported that the incorporation of tissue factor (TF) into cell-derived microvesicles (MVs) is regulated by the phosphorylation of the cytoplasmic domain of TF. Since the cytoskeletal protein filamin-A is known to bind to the cytoplasmic domain of TF in a phosphorylation-dependent manner, the involvement of filamin-A in the incorporation of TF into MVs was examined. Endothelial cells were transfected to express TF, whereas MDA-MB-231 cells were used to examine endogenously expressed TF. MV release was induced by activating protease-activated receptor-2 (PAR2). Partial suppression of filamin-A expression using two different filamin-A siRNA sequences resulted in significant reductions in the incorporation of TF antigen into MVs as determined by TF-ELISA and western blot analysis, and was reflected in reduced thrombin-generation and FXa-generation capacities of these MVs. Deletion of the cytoplasmic domain of TF also resulted in reduced incorporation of TF into MVs, whereas the suppression of filamin-A expression had no additional effect on the incorporation of truncated TF into MVs. Partial suppression of filamin-A expression had no effect on the number and size distribution of the released MVs. However, >90% suppression of filamin-A expression resulted in increased MV release, possibly as a result of increased instability of the plasma membrane and underlying cytoskeleton. In conclusion, the presence of filamin-A appears to be essential for the incorporation of TF into MVs following PAR2 activation, but is not required for the process of MV formation and release following PAR2 activation.

Keywords

Tissue factor - filamin-A - microvesicles - microparticles - endothelial cells

• References

• 1 Kushak RI, Nestoridi E, Lambert J. et al. Detached endothelial cells and microparticles as sources of tissue factor activity.. Thromb Res 2005; 116: 409-419.
• 2 Shet AS, Aras O, Gupta K. et al. Sickle blood contains tissue factor-positive microparticles derived from endothelial cells and monocytes.. Blood 2003; 102: 2678-2683.
• 3 Del Conde I, Shrimpton CN, Thiagarajan P. et al. Tissue-factor-bearing microparticles arise from lipid rafts and fuse with activated platelets to initiate coagulation.. Blood 2005; 106: 1604-1611.
• 4 Schecter AD, Spirn B, Rossikhina M. et al. Release of active tissue factor by human arterial smooth muscle cells.. Circ Res 2000; 87: 126-132.
• 5 Zwicker JI, Liebman HA, Neuberg D. et al. Tumor-derived tissue factor-bearing microparticles are associated with venous thromboembolic events in malignancy.. Clin Cancer Res 2009; 15: 6830-6840.
• 6 Mueller BM, Ruf W. Requirement for binding of catalytically active factor Vila in tissue factor-dependent experimental metastasis.. J Clin Invest 1998; 10: 1372-1378.
• 7 Ahamed J, Ruf W. Protease-activated receptor 2-dependent phosphorylation of the tissue factor cytoplasmic domain.. J Biol Chem 2004; 279: 23038-23044.
• 8 Collier ME, Ettelaie C. Regulation of the incorporation of tissue factor into microparticles by serine phosphorylation of the cytoplasmic domain of tissue factor.. J Biol Chem 2011; 286: 11977-11984.
• 9 Sapet C, Simoncini S, Loriod B. et al. Thrombin-induced endothelial micropar-ticle generation: identification of a novel pathway involving ROCK-II activation by caspase-2.. Blood 2006; 108: 1868-1876.
• 10 Morel O, Jesel L, Freyssinet JM. et al. Cellular mechanisms underlying the formation of circulating microparticles.. Arterioscler Thromb Vasc Biol 2011; 31: 15-26.
• 11 Nakamura F, Stossel TP, Hartwig JH. The filamins: organizers of cell structure and function.. Cell Adh Migr 2011; 5: 160-169.
• 12 Gorlin JB, Yamin R, Egan S. et al. Human endothelial actin-binding protein (ABP-280, nonmuscle filamin): a molecular leaf spring.. J Cell Biol 1990; 111: 1089-1105.
• 13 Ott I, Fischer EG, Miyagi Y. et al. A role for tissue factor in cell adhesion and migration mediated by interaction with actin-binding protein 280.. J Cell Biol 1998; 140: 1241-1253.
• 14 Collier ME, Ettelaie C. Induction of endothelial cell proliferation by recombi-nant and microparticle-tissue factor involves beta1-integrin and extracellular signal regulated kinase activation.. Arterioscler Thromb Vasc Biol 2010; 30: 1810-1817.
• 15 Ettelaie C, Su S, Li C. et al. Tissue factor-containing microparticles released from mesangial cells in response to high glucose and AGE induce tube formation in microvascular cells.. Microvasc Res 2008; 76: 152-160.
• 16 Lin R, Karpa K, Kabbani N, Goldman-Rakic P, Levenson R. Dopamine D2 and D3 receptors are linked to the actin cytoskeleton via interaction with filamin A.. Proc Natl Acad Sci USA 2001; 98: 5258-5263.
• 17 Li M, Bermak JC, Wang ZW. et al. Modulation of dopamine D(2) receptor signaling by actin-binding protein (ABP-280).. Mol Pharmacol 2000; 57: 446-452.
• 18 Mammoto A, Huang S, Ingber DE. Filamin links cell shape and cytoskeletal structure to Rho regulation by controlling accumulation of p190RhoGAP in lipid rafts.. J Cell Sci 2007; 120: 456-467.
• 19 Onoprishvili I, Andria ML, Kramer HK, Ancevska-Taneva N, Hiller JM, Simon EJ. Interaction between the mu opioid receptor and filamin A is involved in receptor regulation and trafficking.. Mol Pharmacol 2003; 64: 1092-1100.
• 20 Seck T, Baron R, Horne WC. Binding of filamin to the C-terminal tail of the calcitonin receptor controls recycling.. J Biol Chem 2003; 278: 10408-10416.
• 21 Jy W, Jimenez JJ, Mauro LM. et al. Agonist-induced capping of adhesion proteins and microparticle shedding in cultures of human renal microvascular en- dothelial cells.. Endothelium 2002; 9: 179-189.
• 22 Peña E, Arderiu G, Badimon L. Subcellular localization of tissue factor and human coronary artery smooth muscle cell migration.. J Thromb Haemost 2012; 10: 2373-2382.
• 23 Davizon P, Munday AD, López JA. Tissue factor, lipid rafts, and microparticles.. Semin Thromb Hemost 2010; 36: 857-864.
• 24 Mandal SK, Pendurthi UR, Rao LV. Tissue factor trafficking in fibroblasts: involvement of protease-activated receptor-mediated cell signaling.. Blood 2007; 110: 161-170.
• 25 Bona R, Lee E, Rickles F. Tissue factor apoprotein: intracellular transport and expression in shed membrane vesicles.. Thromb Res 1987; 48: 487-500.
• 26 Feng S, Lu X, Kroll MH. Filamin A binding stabilizes nascent glycoprotein Ibalpha trafficking and thereby enhances its surface expression.. J Biol Chem 2005; 280: 6709-6715.
• 27 Kanaji T, Ware J, Okamura T. et al. GPIba regulates platelet size by controlling the subcellular localization of filamin.. Blood 2012; 119: 2906-2913.
• 28 Legler DF, Doucey MA, Schneider P. et al. Differential insertion of GPI-an-chored GFPs into lipid rafts of live cells.. FASEB J 2005; 19: 73-75.
• 29 Brown D, Waneck GL. Glycosyl-phosphatidylinositol-anchored membrane proteins.. J Am Soc Nephrol 1992; 3: 895-906.
• 30 Nasu J, Mizuno M, Uesu T. et al. Cytokine-stimulated release of decay-accelerating factor (DAF;CD55) from HT-29 human intestinal epithelial cells.. Clin Exp Immuno 1998; 113: 379-385.
• 31 Cunningham CC, Gorlin JB, Kwiatkowski DJ. et al. Actin-binding protein requirement for cortical stability and efficient locomotion.. Science 1992; 255: 325-327.
• 32 Jimenez JJ, Jy W, Mauro LM. et al. Endothelial cells release phenotypically and quantitatively distinct microparticles in activation and apoptosis.. Thromb Res 2003; 109: 175-180.