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Expression of the endocannabinoid system in the bi-potential HEL cell line: commitment to the megakaryoblastic lineage by 2-arachidonoylglycerol

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Abstract

The role of the endocannabinoid system in haematopoietic cells is not completely understood. We investigated whether human erythroleukemia (HEL) cells were able to bind, metabolise and transport the main endocannabinoids, anandamide (AEA) and 2-arachidonoylglycerol (2-AG). We also investigated whether AEA or 2-AG could modulate HEL differentiation. Although able to internalise both endocannabinoids, HEL cells had the machinery to metabolise 2-AG only, since they were devoid of the enzymes needed to synthesise and degrade AEA. Nonetheless, the intracellular transport of exogenous AEA might be required to activate the vanilloid receptors, with yet unknown implications for vascular biology. On the contrary, 2-AG appeared to play a role in lineage determination. Indeed, 2-AG itself drove HEL cells towards megakaryocytic differentiation, as it enhanced expression of β3 integrin subunit, a megakaryocyte/platelet surface antigen, and glycoprotein VI, a late marker of megakaryocytes; in parallel, it reduced the amount of messenger RNA encoding for glycophorin A, a marker of erythroid phenotype. All these effects were mediated by activation of CB2 cannabinoid receptors that triggered an extracellular signal-regulated kinase-dependent signalling cascade. In addition, classical inducers of megakaryocyte differentiation reduced 2-AG synthesis (although they did not affect the binding efficiency of CB2 receptors), suggesting that levels of this endocannabinoid may be critical for committing HEL cells towards the megakaryocytic lineage.

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Abbreviations

HEL:

human erythroleukaemia

AEA:

anandamide

2-AG:

2-arachidonoylglycerol

CB1 :

type-1 cannabinoid receptor

CB2 :

type-2 cannabinoid receptor

NAPE-PLD:

N-acyl-phosphatidylethanolamines-hydrolyzing phospholipase D

FAAH:

fatty acid amide hydrolase

TRPV1:

transient receptor potential channel vanilloid receptor subunit 1

DAGL:

sn-1-specific diacylglycerol lipase

MAGL:

monoacylglycerol lipase

RTX:

resinferatoxin

CP55,940:

5-(1,10-dimethyheptyl)-2-[1R,5R-hydroxy-2R-(3-hydroxypropyl)-cyclohexyl]phenol

SR141716:

N-piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-3-pyrazole-carboxa-mide

SR144528:

[N-[(1S)-endo-1,3,3-trimethy-1-bicyclo [2.2.1]-heptan-2-yl]5-(4-choro-3-methyl-phenyl)-1-(4-methyl-benzyl)-pyrazole-3-carboxamide]

OMDM1:

(R)-N-oleoyl-(1′-hydroxybenzyl)-2′-ethanolamin

NarPE:

(N-arachidonoyl-phosphatidylethanolamine

2-OG:

2-oleoyl-glycerol

DAG:

sn-1-stearoyl-2arachidonoyl-glycerol

References

  1. Bari M, Battista N, Fezza F, Gasperi V, Maccarrone M (2006) New insights into endocannabinoid degradation and its therapeutic potential. Mini Rev Med Chem 6:257–268

    Article  PubMed  CAS  Google Scholar 

  2. Di Marzo V (2006) A brief history of cannabinoid and endocannabinoid pharmacology as inspired by the work of British scientists. Trends Pharmacol Sci 27:134–140

    Article  PubMed  CAS  Google Scholar 

  3. Egertova M, Cravatt BF, Elphick MR (2003) Comparative analysis of fatty acid amide hydrolase and CB1 cannabinoid receptor expression in the mouse brain: evidence of a widespread role for fatty acid amide hydrolase in regulation of endocannabinoid signaling. Neuroscience 119:481–496

    Article  PubMed  CAS  Google Scholar 

  4. Van Sickle MD, Duncan M, Kingsley PJ, Mouihate A, Urbani P, Mackie K, Stella N, Makriyannis A, Piomelli D, Davison JS, Marnett LJ, Di Marzo V, Pittman QJ, Patel KD, Sharkey KA (2005) Identification and functional characterization of brain stem cannabinoid CB2 receptors. Science 310:329–332

    Article  PubMed  CAS  Google Scholar 

  5. Okamoto Y, Morishita J, Tsuboi K, Tonai T, Ueda N (2004) Molecular characterization of a phospholipase D generating anandamide and Its congeners. J Biol Chem 279:5298–5305

    Article  PubMed  CAS  Google Scholar 

  6. Battista N, Gasperi V, Fezza F, Maccarrone M (2005) The anandamide membrane transporter and the therapeutic implications of its inhibition. Therapy 2:141–150

    Article  CAS  Google Scholar 

  7. McKinney MK, Cravatt BF (2005) Structure and function of fatty acid amide hydrolase. Annu Rev Biochem 74:411–432

    Article  PubMed  CAS  Google Scholar 

  8. Van der Stelt M, Di Marzo V (2004) Endovanilloids. Putative endogenous ligands of transient receptor potential vanilloid 1 channels. Eur J Biochem 271:1827–1834

    Article  CAS  Google Scholar 

  9. Bisogno T, Howell F, Williams G, Minassi A, Cascio MG, Ligresti A, Matias I, Schiano-Moriello A, Paul P, Williams EJ, Gangadharan U, Hobbs C, Di Marzo V, Doherty P (2003) Cloning of the first sn1-DAG lipases points to the spatial and temporal regulation of endocannabinoid knockout in the brain. J Cell Biol 163:463–468

    Article  PubMed  CAS  Google Scholar 

  10. Dinh TP, Carpenter D, Leslie FM, Freund TF, Katona I, Sensi SL, Kathuria S, Piomelli D (2002) Brain monoglyceride lipase participating in endocannabinoid inactivation. Proc Natl Acad Sci USA 99:10819–10824

    Article  PubMed  CAS  Google Scholar 

  11. Hermann A, Kaczocha M, Deutsch DG (2006) 2-Arachidonoylglycerol (2-AG) membrane transport: history and outlook. AAPS J 8:E409–412

    PubMed  Google Scholar 

  12. Piomelli D (2003) The molecular logic of endocannabinoid signaling. Nat Rev Neurosci 4:873–884

    Article  PubMed  CAS  Google Scholar 

  13. Jorda MA, Rayman N, Tas M, Verbakel SE, Battista N, van Lom K, Lowenberg B, Maccarrone M, Delwel R (2004) The peripheral cannabinoid receptor Cb2, frequently expressed on AML blasts, either induces a neutrophilic differentiation block or confers abnormal migration properties in a ligand-dependent manner. Blood 104:526–534

    Article  CAS  Google Scholar 

  14. Pacher P, Batkai S, Kunos G (2005) Cardiovascular pharmacology of cannabinoids. Handb Exp Pharmacol 168:599–625

    Article  PubMed  CAS  Google Scholar 

  15. Maccarrone M, Bari M, Menichelli A, Del Principe D, Finazzi-Agrò A (1999) Anandamide activates human platelets through a pathway independent of the arachidonate cascade. FEBS Lett 447:277–282

    Article  PubMed  CAS  Google Scholar 

  16. Maccarrone M, Bari M, Del Principe D, Finazzi-Agrò A (2003) Activation of human platelets by 2 arachidonoylglycerol is enhanced by serotonin. Thromb Haemost 89:340–347

    PubMed  CAS  Google Scholar 

  17. Maccarrone M, Bari M, Battista N, Finazzi-Agrò A (2002) Estrogen stimulates arachidonoylethanolamide release from human endothelial cells and platelet activation. Blood 100:4040–4048

    Article  PubMed  CAS  Google Scholar 

  18. Galve-Roperh I, Aguado T, Rueda D, Velasco G, Guzman M (2006) Endocannabinoids: a new family of lipid mediators involved in the regulation of neural cell development. Curr Pharm Des 12:2319–2325

    Article  PubMed  CAS  Google Scholar 

  19. Matias I, Gonthier MP, Orlando P, Martiadis V, De Petrocellis L, Cervino C, Petrosino S, Hoareau L, Festy F, Pasquali R, Roche R, Maj M, Pagotto U, Monteleone P, Di Marzo V (2006) Regulation, function, and dysregulation of endocannabinoids in models of adipose and beta-pancreatic cells and in obesity and hyperglycemia. J Clin Endocrinol Metab 91:3171–3180

    Article  PubMed  CAS  Google Scholar 

  20. Jorda MA, Lowenberg B, Delwel R (2003) The peripheral cannabinoid receptor Cb2, a novel oncoprotein, induces a reversible block in neutrophilic differentiation. Blood 101:1336–1343

    Article  PubMed  CAS  Google Scholar 

  21. Akashi K, Traver D, Miyamoto T, Weissman IL (2000) A clonogenic common myeloid progenitor that gives rise to all myeloid lineages. Nature 404:193–197

    Article  PubMed  CAS  Google Scholar 

  22. Papayannopoulou T, Nakamoto B, Kurachi S, Nelson R (1987) Analysis of the erythroid phenotype of HEL cells: clonal variation and the effect of inducers. Blood 70:1764–1772

    PubMed  CAS  Google Scholar 

  23. Tabilio A, Rosa JP, Testa U, Kieffer N, Nurden AT, Del Canizo MC, Breton-Gorius J, Vainchenker W (1984) Expression of platelet membrane glycoproteins and alpha-granule proteins by a human erythroleukemia cell line (HEL). EMBO J 3:453–459

    PubMed  CAS  Google Scholar 

  24. Savini I, Catani MV, Arnone R, Rossi A, Frega G, Del Principe D, Avigliano L (2007) Translational control of the ascorbic acid transporter SVCT2 in human platelets. Free Radic Biol Med 42:608–616

    Article  PubMed  CAS  Google Scholar 

  25. Maccarrone M, Barboni B, Paradisi A, Bernabo N, Gasperi V, Pistilli MG, Fezza F, Lucidi P, Mattioli M (2005) Characterization of the endocannabinoid system in boar spermatozoa and implications for sperm capacitation and acrosome reaction. J Cell Sci 118:4393–4404

    Article  PubMed  CAS  Google Scholar 

  26. Hillard CJ, Edgemond WS, Jarrahian A, Campbell WB (1997) Accumulation of N-arachidonoylethanolamine (anandamide) into cerebellar granule cells occurs via facilitated diffusion. J Neurochem 69:631–638

    Article  PubMed  CAS  Google Scholar 

  27. de Lago E, Ligresti A, Ortar G, Morera E, Cabranes A, Pryce G, Bifulco M, Baker D, Fernandez-Ruiz J, Di Marzo V (2004) In vivo pharmacological actions of two novel inhibitors of anandamide cellular uptake. Eur J Pharmacol 484:249–257

    Article  PubMed  CAS  Google Scholar 

  28. Pertwee RG (1997) Pharmacology of cannabinoid CB1 and CB2 receptors. Pharmacol Ther 74:129–180

    Article  PubMed  CAS  Google Scholar 

  29. Sugiura T, Kondo S, Kishimoto S, Miyashita T, Nakane S, Kodaka T, Suhara Y, Takayama H, Waku K (2000) Evidence that 2-arachidonoylglycerol but not N-palmitoylethanolamine or anandamide is the physiological ligand for the cannabinoid CB2 receptor. Comparison of the agonistic activities of various cannabinoid receptor ligands in HL-60 cells. J Biol Chem 275:605–612

    Article  PubMed  CAS  Google Scholar 

  30. Maccarrone M, Bari M, Menichelli A, Giuliani E, Del Principe D, Finazzi Agrò A (2001) Human platelets bind and degrade 2-arachidonoylglycerol, which activates these cells through a cannabinoid receptor. Eur J Biochem 268:819–825

    Article  PubMed  CAS  Google Scholar 

  31. Marshall CJ (1995) Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. Cell 80:179–185

    Article  PubMed  CAS  Google Scholar 

  32. Wu H, Shen HW, Wu TF, Brass LF, Sung KC (2002) Extracellular signal-regulated kinases and g protein-coupled receptors in megakaryocytic human erythroleukemia cells: selective activation, differential regulation, and dissociation from mitogenesis. J Pharmacol Exp Ther 300:339–345

    Article  PubMed  CAS  Google Scholar 

  33. Woessmann W, Zwanzger D, Borkhardt A (2004) ERK signaling pathway is differentially involved in erythroid differentiation of K562 cells depending on time and the inducing agent. Cell Biol Int 28:403–410

    Article  PubMed  CAS  Google Scholar 

  34. Siemens J, Zhou S, Piskorowski R, Nikai T, Lumpkin EA, Basbaum AI, King D, Julius D (2006) Spider toxins activate the capsaicin receptor to produce inflammatory pain. Nature 444:208–212

    Article  PubMed  CAS  Google Scholar 

  35. Rabellino EM, Levene RB, Leung LL, Nachman RL (1981) Human megakaryocytes. II. Expression of platelet proteins in early marrow megakaryocytes. J Exp Med 154:88–100

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This work was supported by grants from Ministero dell’Università e della Ricerca (PRIN 2004), from Ministero della Salute (R.C. 2005), from Fondazione della Cassa di Risparmio di Teramo (TERCAS contract 2005) and from Regione Piemonte -Ricerca Sanitaria Finalizzata 2004 (AB).

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Authors and Affiliations

  1. Department of Experimental Medicine & Biochemical Sciences, University of Rome Tor Vergata, Rome, Italy

    Maria Valeria Catani, Filomena Fezza, Alessandro Finazzi-Agrò & Luciana Avigliano

  2. European Center for Brain Research (CERC)/IRCCS S. Lucia Foundation, Rome, Italy

    Filomena Fezza, Valeria Gasperi, Nicoletta Pasquariello & Mauro Maccarrone

  3. Department of Medical Sciences, University of Eastern Piedmont “A. Avogadro”, Novara, Italy

    Samantha Baldassarri, Alessandra Bertoni & Fabiola Sinigaglia

  4. Department of Biomedical Sciences, University of Teramo, Piazza A. Moro 45, 64100, Teramo, Italy

    Valeria Gasperi, Nicoletta Pasquariello & Mauro Maccarrone

  5. Center of Biotechnology for Applied Medical Research (BRMA), Novara, Italy

    Alessandra Bertoni & Fabiola Sinigaglia

Authors
  1. Maria Valeria Catani
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  2. Filomena Fezza
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Correspondence to Mauro Maccarrone.

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Maria Valeria Catani and Filomena Fezza contributed equally to the study.

Luciana Avigliano and Mauro Maccarrone are equally senior authors.

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Catani, M.V., Fezza, F., Baldassarri, S. et al. Expression of the endocannabinoid system in the bi-potential HEL cell line: commitment to the megakaryoblastic lineage by 2-arachidonoylglycerol. J Mol Med 87, 65–74 (2009). https://doi.org/10.1007/s00109-008-0406-3

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