Subscribe to RSS
DOI: 10.1055/s-0039-1693737
All-Trans Retinoic Acid Impairs Platelet Function and Thrombus Formation and Inhibits Protein Kinase CßI/δ Phosphorylation
Funding This research was supported by National Natural Science Foundation of China (grant no. 81400082, 81641151, and 81700178), the Natural Science Foundation of Jiangsu Province (grant no. BK20140219 and BK20170259), the funding for the Distinguished Professorship Program of Jiangsu Province, the Shuangchuang Project of Jiangsu Province, the Six Talent Peaks Project of Jiangsu Province (WSN-133), the 333 projects of Jiangsu Province (BRA2017542), the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (18KJA320010 and 17KJA320008), Jiangsu Province's Key Provincial Talents Program (ZDRCA2016054), Jiangsu Province’s Graduate Scientific Research Innovation Program (KYCX18-2186, KYCX19-2231, KYCX19-2234) and the National Health and Medical Research Council of Australia.Publication History
14 March 2019
12 June 2019
Publication Date:
01 August 2019 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/201910/lookinside/thumbnails/10-1055-s-0039-1693737_190145-1.jpg)
Abstract
All-trans retinoic acid (ATRA) is widely used for induction of complete remission in patients with acute promyelocytic leukemia (APL). ATRA also regulates protein kinase C (PKC) activity. Therapeutic use of ATRA reportedly interferes with hemostatic function in APL patients, including effects on coagulation or other vascular cells, although effects of ATRA on platelets remain unclear. This study aims to investigate the effect of therapeutic-relevant doses of ATRA on platelet function. Human platelets were preincubated with ATRA (0–20 μM) for 1 hour at 37°C, followed by analysis of aggregation, granule secretion, receptor expression by flow cytometry, platelet spreading, or clot retraction. Additionally, ATRA (10 mg/kg) was injected intraperitoneally into mice and tail bleeding time and arterial thrombus formation were evaluated. ATRA inhibited platelet aggregation and adenosine triphosphate release induced by collagen (5 μg/mL) or thrombin (0.05 U/mL) in a dose-dependent manner without affecting P-selectin expression or surface levels of glycoprotein (GP) Ibα, GPVI, or αIIbβ3. ATRA-treated platelets demonstrated reduced spreading on immobilized fibrinogen or collagen and reduced thrombin-induced clot retraction together with reduced phosphorylation of Syk and PLCγ2. In addition, ATRA-treated mice displayed significantly impaired hemostasis and arterial thrombus formation in vivo. Further, in platelets stimulated with either collagen-related peptide or thrombin, ATRA selectively inhibited phosphorylation of PKCßI (Ser661) and PKCδ (Thr505), but not PKCα or PKCßII phosphorylation (Thr638/641). In conclusion, ATRA inhibits platelet function and thrombus formation, possibly involving direct or indirect inhibition of PKCßI/δ, indicating that ATRA might be beneficial for the treatment of thrombotic or cardiovascular diseases.
Keywords
all-trans retinoic acid - platelet - hemostasis - thrombus formation - protein kinase CßI/δ* These authors contributed equally to this study.
-
References
- 1 Qiao JL, Shen Y, Gardiner EE, Andrews RK. Proteolysis of platelet receptors in humans and other species. Biol Chem 2010; 391 (08) 893-900
- 2 Qiao J, Arthur JF, Gardiner EE, Andrews RK, Zeng L, Xu K. Regulation of platelet activation and thrombus formation by reactive oxygen species. Redox Biol 2018; 14: 126-130
- 3 Li Z, Delaney MK, O'Brien KA, Du X. Signaling during platelet adhesion and activation. Arterioscler Thromb Vasc Biol 2010; 30 (12) 2341-2349
- 4 Durrant TN, van den Bosch MT, Hers I. Integrin αIIbβ3 outside-in signaling. Blood 2017; 130 (14) 1607-1619
- 5 Siddikuzzaman GC, Guruvayoorappan C, Berlin Grace VM. All trans retinoic acid and cancer. Immunopharmacol Immunotoxicol 2011; 33 (02) 241-249
- 6 Napoli JL. Retinoic acid biosynthesis and metabolism. FASEB J 1996; 10 (09) 993-1001
- 7 Allenby G, Janocha R, Kazmer S, Speck J, Grippo JF, Levin AA. Binding of 9-cis-retinoic acid and all-trans-retinoic acid to retinoic acid receptor-alpha, receptor-beta, and receptor-gamma - retinoic acid receptor-gamma binds all-trans-retinoic acid preferentially over 9-cis-retinoic acid. J Biol Chem 1994; 269 (24) 16689-16695
- 8 Cicconi L, Lo-Coco F. Current management of newly diagnosed acute promyelocytic leukemia. Ann Oncol 2016; 27 (08) 1474-1481
- 9 Wang ZY, Chen Z. Acute promyelocytic leukemia: from highly fatal to highly curable. Blood 2008; 111 (05) 2505-2515
- 10 Dos Santos GA, Kats L, Pandolfi PP. Synergy against PML-RARa: targeting transcription, proteolysis, differentiation, and self-renewal in acute promyelocytic leukemia. J Exp Med 2013; 210 (13) 2793-2802
- 11 Warrell Jr RP, de Thé H, Wang ZY, Degos L. Acute promyelocytic leukemia. N Engl J Med 1993; 329 (03) 177-189
- 12 Falanga A, Iacoviello L, Evangelista V. , et al. Loss of blast cell procoagulant activity and improvement of hemostatic variables in patients with acute promyelocytic leukemia administered all-trans-retinoic acid. Blood 1995; 86 (03) 1072-1081
- 13 Dombret H, Scrobohaci ML, Ghorra P. , et al. Coagulation disorders associated with acute promyelocytic leukemia: corrective effect of all-trans retinoic acid treatment. Leukemia 1993; 7 (01) 2-9
- 14 Kawai Y, Watanabe K, Kizaki M. , et al. Rapid improvement of coagulopathy by all-trans retinoic acid in acute promyelocytic leukemia. Am J Hematol 1994; 46 (03) 184-188
- 15 Federici AB, Falanga A, Lattuada A, Di Rocco N, Barbui T, Mannucci PM. Proteolysis of von Willebrand factor is decreased in acute promyelocytic leukaemia by treatment with all-trans-retinoic acid. Br J Haematol 1996; 92 (03) 733-739
- 16 Barbui T, Finazzi G, Falanga A. The impact of all-trans-retinoic acid on the coagulopathy of acute promyelocytic leukemia. Blood 1998; 91 (09) 3093-3102
- 17 Rondina MT, Freitag M, Pluthero FG. , et al. Non-genomic activities of retinoic acid receptor alpha control actin cytoskeletal events in human platelets. J Thromb Haemost 2016; 14 (05) 1082-1094
- 18 Arthur JF, Qiao J, Shen Y. , et al. ITAM receptor-mediated generation of reactive oxygen species in human platelets occurs via Syk-dependent and Syk-independent pathways. J Thromb Haemost 2012; 10 (06) 1133-1141
- 19 Qiao J, Wu X, Luo Q. , et al. NLRP3 regulates platelet integrin αIIbβ3 outside-in signaling, hemostasis and arterial thrombosis. Haematologica 2018; 103 (09) 1568-1576
- 20 Qiao J, Wu Y, Liu Y. , et al. Busulfan triggers intrinsic mitochondrial-dependent platelet apoptosis independent of platelet activation. Biol Blood Marrow Transplant 2016; 22 (09) 1565-1572
- 21 Luo Q, Wei G, Wu X. , et al. Platycodin D inhibits platelet function and thrombus formation through inducing internalization of platelet glycoprotein receptors. J Transl Med 2018; 16 (01) 311
- 22 Asmis L, Tanner FC, Sudano I, Lüscher TF, Camici GG. DMSO inhibits human platelet activation through cyclooxygenase-1 inhibition. A novel agent for drug eluting stents?. Biochem Biophys Res Commun 2010; 391 (04) 1629-1633
- 23 Lehuu B, Curtis-Prior PB. Effects of dimethyl sulphoxide (DMSO) on aggregation of human blood platelets. J Pharm Pharmacol 1987; 39 (01) 62-63
- 24 Gardiner EE, Andrews RK. Structure and function of platelet receptors initiating blood clotting. Adv Exp Med Biol 2014; 844: 263-275
- 25 Bennett JS. Structure and function of the platelet integrin alphaIIbbeta3. J Clin Invest 2005; 115 (12) 3363-3369
- 26 Flevaris P, Stojanovic A, Gong H, Chishti A, Welch E, Du X. A molecular switch that controls cell spreading and retraction. J Cell Biol 2007; 179 (03) 553-565
- 27 Suzuki-Inoue K, Hughes CE, Inoue O. , et al. Involvement of Src kinases and PLCgamma2 in clot retraction. Thromb Res 2007; 120 (02) 251-258
- 28 Radominska-Pandya A, Chen G, Czernik PJ. , et al. Direct interaction of all-trans-retinoic acid with protein kinase C (PKC). Implications for PKC signaling and cancer therapy. J Biol Chem 2000; 275 (29) 22324-22330
- 29 Harper MT, Poole AW. Diverse functions of protein kinase C isoforms in platelet activation and thrombus formation. J Thromb Haemost 2010; 8 (03) 454-462
- 30 Ishii H, Horie S, Kizaki K, Kazama M. Retinoic acid counteracts both the downregulation of thrombomodulin and the induction of tissue factor in cultured human endothelial cells exposed to tumor necrosis factor. Blood 1992; 80 (10) 2556-2562
- 31 Dittman WA, Nelson SC, Greer PK, Horton ET, Palomba ML, McCachren SS. Characterization of thrombomodulin expression in response to retinoic acid and identification of a retinoic acid response element in the human thrombomodulin gene. J Biol Chem 1994; 269 (24) 16925-16932
- 32 van Giezen JJ, Boon GI, Jansen JW, Bouma BN. Retinoic acid enhances fibrinolytic activity in-vivo by enhancing tissue type plasminogen activator (t-PA) activity and inhibits venous thrombosis. Thromb Haemost 1993; 69 (04) 381-386
- 33 Falanga A, Consonni R, Marchetti M. , et al. Cancer procoagulant in the human promyelocytic cell line NB4 and its modulation by all-trans-retinoic acid. Leukemia 1994; 8 (01) 156-159
- 34 Rivera J, Lozano ML, Navarro-Núñez L, Vicente V. Platelet receptors and signaling in the dynamics of thrombus formation. Haematologica 2009; 94 (05) 700-711
- 35 Murugappan S, Tuluc F, Dorsam RT, Shankar H, Kunapuli SP. Differential role of protein kinase C delta isoform in agonist-induced dense granule secretion in human platelets. J Biol Chem 2004; 279 (04) 2360-2367
- 36 Chari R, Getz T, Nagy Jr B. , et al. Protein kinase C[delta] differentially regulates platelet functional responses. Arterioscler Thromb Vasc Biol 2009; 29 (05) 699-705