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Abstract
Background: Cannabinoid receptors have been detected in human gliomas and cannabinoids have been proposed as novel drug candidates in the treatment of brain tumors.
Aims: To test the in vitro antitumor activity of COR167, a novel cannabinoid CB2-selective agonist displaying a high binding affinity for human CB2 receptors, on tumor cells isolated from human glioblastoma multiforme and anaplastic astrocytoma. Methods: Glioma cell cultures were established from two glioblastoma multiforme and two anaplastic astrocytomas. Proliferation was measured in the presence or absence of COR167 with a bromodeoxyuridine (BrdU) cell proliferation ELISA assay. CB2 receptor expression was detected by western blotting. Apoptosis was assessed with phycoerythrin (PE) annexin V flow cytometry kit. TGF-beta 1 and 2 levels were analyzed in culture supernatants with commercial ELISAs. Results: COR167 was found to significantly reduce the proliferation of both glioblastoma and anaplastic astrocytoma in a dose-dependent manner at lower doses than other known, less specific CB2 agonists. This activity is independent of apoptosis and is associated with a significant reduction of TGF-beta 1 and 2 levels in supernatants of glioma cell cultures. Conclusion: These findings add to the role of cannabinoid CB2 receptor as a possible pharmacological target to counteract glial tumor growth and encourage further work to explore any other pharmacological effect of this novel CB2 agonist useful in the treatment of human gliomas.Keywords: Glioma therapy, cannabinoids, CB2 receptor, TGF-beta, glioblastoma multiforme, anaplastic astrocytoma.
[1]
Cohen, A.L.; Colman, H. Glioma biology and molecular markers Cancer Treat. Res, 2015, 163, 15-30.
[http://dx.doi.org/10.1007/978-3-319-12048-5_2] [PMID: 25468223]
[http://dx.doi.org/10.1007/978-3-319-12048-5_2] [PMID: 25468223]
[2]
Krex, D.; Klink, B.; Hartmann, C.; von Deimling, A.; Pietsch, T.; Simon, M.; Sabel, M.; Steinbach, J.P.; Heese, O.; Reifenberger, G.; Weller, M.; Schackert, G. Long-term survival with glioblastoma multiforme. Brain, 2007, 130(Pt 10), 2596-2606.
[http://dx.doi.org/10.1093/brain/awm204] [PMID: 17785346]
[http://dx.doi.org/10.1093/brain/awm204] [PMID: 17785346]
[3]
Binder, D.C.; Davis, A.A.; Wainwright, D.A. Immunotherapy for cancer in the central nervous system: Current and future directions. OncoImmunology, 2015, 5(2) e1082027
[http://dx.doi.org/10.1080/2162402X.2015.1082027] [PMID: 27057463]
[http://dx.doi.org/10.1080/2162402X.2015.1082027] [PMID: 27057463]
[4]
Alifieris, C.; Trafalis, D.T. Glioblastoma multiforme: Pathogenesis and treatment. Pharmacol. Ther., 2015, 152, 63-82.
[http://dx.doi.org/10.1016/j.pharmthera.2015.05.005] [PMID: 25944528]
[http://dx.doi.org/10.1016/j.pharmthera.2015.05.005] [PMID: 25944528]
[5]
Folkman, J. Angiogenesis: an organizing principle for drug discovery? Nat. Rev. Drug Discov., 2007, 6(4), 273-286.
[http://dx.doi.org/10.1038/nrd2115] [PMID: 17396134]
[http://dx.doi.org/10.1038/nrd2115] [PMID: 17396134]
[6]
Held-Feindt, J.; Dörner, L.; Sahan, G.; Mehdorn, H.M.; Mentlein, R. Cannabinoid receptors in human astroglial tumors. J. Neurochem., 2006, 98(3), 886-893.
[http://dx.doi.org/10.1111/j.1471-4159.2006.03911.x] [PMID: 16893424]
[http://dx.doi.org/10.1111/j.1471-4159.2006.03911.x] [PMID: 16893424]
[7]
Ellert-Miklaszewska, A.; Grajkowska, W.; Gabrusiewicz, K.; Kaminska, B.; Konarska, L. Distinctive pattern of cannabinoid receptor type II (CB2) expression in adult and pediatric brain tumors. Brain Res., 2007, 1137(1), 161-169.
[http://dx.doi.org/10.1016/j.brainres.2006.12.060] [PMID: 17239827]
[http://dx.doi.org/10.1016/j.brainres.2006.12.060] [PMID: 17239827]
[8]
Carracedo, A.; Lorente, M.; Egia, A.; Blázquez, C.; García, S.; Giroux, V.; Malicet, C.; Villuendas, R.; Gironella, M.; González-Feria, L.; Piris, M.A.; Iovanna, J.L.; Guzmán, M.; Velasco, G. The stress-regulated protein p8 mediates cannabinoid-induced apoptosis of tumor cells. Cancer Cell, 2006, 9(4), 301-312.
[http://dx.doi.org/10.1016/j.ccr.2006.03.005] [PMID: 16616335]
[http://dx.doi.org/10.1016/j.ccr.2006.03.005] [PMID: 16616335]
[9]
Blázquez, C.; González-Feria, L.; Álvarez, L.; Haro, A.; Casanova, M.L.; Guzmán, M. Cannabinoids inhibit the vascular endothelial growth factor pathway in gliomas. Cancer Res., 2004, 64(16), 5617-5623.
[http://dx.doi.org/10.1158/0008-5472.CAN-03-3927] [PMID: 15313899]
[http://dx.doi.org/10.1158/0008-5472.CAN-03-3927] [PMID: 15313899]
[10]
Samoto, K.; Ikezaki, K.; Ono, M.; Shono, T.; Kohno, K.; Kuwano, M.; Fukui, M. Expression of vascular endothelial growth factor and its possible relation with neovascularization in human brain tumors. Cancer Res., 1995, 55(5), 1189-1193.
[PMID: 7532545]
[PMID: 7532545]
[11]
Schmidt, N.O.; Westphal, M.; Hagel, C.; Ergün, S.; Stavrou, D.; Rosen, E.M.; Lamszus, K. Levels of vascular endothelial growth factor, hepatocyte growth factor/scatter factor and basic fibroblast growth factor in human gliomas and their relation to angiogenesis. Int. J. Cancer, 1999, 84(1), 10-18.
[http://dx.doi.org/10.1002/(SICI)1097-0215(19990219)84:1<10:AID-IJC3>3.0.CO;2-L] [PMID: 9988225]
[http://dx.doi.org/10.1002/(SICI)1097-0215(19990219)84:1<10:AID-IJC3>3.0.CO;2-L] [PMID: 9988225]
[12]
Blázquez, C.; Salazar, M.; Carracedo, A.; Lorente, M.; Egia, A.; González-Feria, L.; Haro, A.; Velasco, G.; Guzmán, M. Cannabinoids inhibit glioma cell invasion by down-regulating matrix metalloproteinase-2 expression. Cancer Res., 2008, 68(6), 1945-1952.
[http://dx.doi.org/10.1158/0008-5472.CAN-07-5176] [PMID: 18339876]
[http://dx.doi.org/10.1158/0008-5472.CAN-07-5176] [PMID: 18339876]
[13]
Casanova, M.L.; Blázquez, C.; Martínez-Palacio, J.; Villanueva, C.; Fernández-Aceñero, M.J.; Huffman, J.W.; Jorcano, J.L.; Guzmán, M. Inhibition of skin tumor growth and angiogenesis in vivo by activation of cannabinoid receptors. J. Clin. Invest., 2003, 111(1), 43-50.
[http://dx.doi.org/10.1172/JCI200316116] [PMID: 12511587]
[http://dx.doi.org/10.1172/JCI200316116] [PMID: 12511587]
[14]
Sánchez, C.; de Ceballos, M.L.; Gomez del Pulgar, T.; Rueda, D.; Corbacho, C.; Velasco, G.; Galve-Roperh, I.; Huffman, J.W.; Ramón y Cajal, S.; Guzmán, M. Inhibition of glioma growth in vivo by selective activation of the CB(2) cannabinoid receptor. Cancer Res., 2001, 61(15), 5784-5789.
[PMID: 11479216]
[PMID: 11479216]
[15]
Hanus, L.; Breuer, A.; Tchilibon, S.; Shiloah, S.; Goldenberg, D.; Horowitz, M.; Pertwee, R.G.; Ross, R.A.; Mechoulam, R.; Fride, E. HU-308: A specific agonist for CB(2), a peripheral cannabinoid receptor. Proc. Natl. Acad. Sci. USA, 1999, 96(25), 14228-14233.
[http://dx.doi.org/10.1073/pnas.96.25.14228] [PMID: 10588688]
[http://dx.doi.org/10.1073/pnas.96.25.14228] [PMID: 10588688]
[16]
Huffman, J.W.; Liddle, J.; Yu, S.; Aung, M.M.; Abood, M.E.; Wiley, J.L.; Martin, B.R. 3-(1′,1′-Dimethylbutyl)-1-deoxy-delta8-THC and related compounds: synthesis of selective ligands for the CB2 receptor. Bioorg. Med. Chem., 1999, 7(12), 2905-2914.
[http://dx.doi.org/10.1016/S0968-0896(99)00219-9] [PMID: 10658595]
[http://dx.doi.org/10.1016/S0968-0896(99)00219-9] [PMID: 10658595]
[17]
Pasquini, S.; Botta, L.; Semeraro, T.; Mugnaini, C.; Ligresti, A.; Palazzo, E.; Maione, S.; Di Marzo, V.; Corelli, F. Investigations on the 4-quinolone-3-carboxylic acid motif. 2. Synthesis and structure-activity relationship of potent and selective cannabinoid-2 receptor agonists endowed with analgesic activity in vivo. J. Med. Chem., 2008, 51(16), 5075-5084.
[http://dx.doi.org/10.1021/jm800552f] [PMID: 18680276]
[http://dx.doi.org/10.1021/jm800552f] [PMID: 18680276]
[18]
Cascio, M.G.; Bolognini, D.; Pertwee, R.G.; Palazzo, E.; Corelli, F.; Pasquini, S.; Di Marzo, V.; Maione, S. In vitro and in vivo pharmacological characterization of two novel selective cannabinoid CB(2) receptor inverse agonists. Pharmacol. Res., 2010, 61(4), 349-354.
[http://dx.doi.org/10.1016/j.phrs.2009.11.011] [PMID: 19961936]
[http://dx.doi.org/10.1016/j.phrs.2009.11.011] [PMID: 19961936]
[19]
Annunziata, P.; Cioni, C.; Mugnaini, C.; Corelli, F. Potent immunomodulatory activity of a highly selective cannabinoid CB2 agonist on immune cells from healthy subjects and patients with multiple sclerosis. J. Neuroimmunol., 2017, 303, 66-74.
[http://dx.doi.org/10.1016/j.jneuroim.2016.12.009] [PMID: 28041663]
[http://dx.doi.org/10.1016/j.jneuroim.2016.12.009] [PMID: 28041663]
[20]
Brunet, J.F.; Pellerin, L.; Magistretti, P.; Villemure, J.G. Cryopreservation of human brain tissue allowing timely production of viable adult human brain cells for autologous transplantation. Cryobiology, 2003, 47(2), 179-183.
[http://dx.doi.org/10.1016/j.cryobiol.2003.08.005] [PMID: 14580852]
[http://dx.doi.org/10.1016/j.cryobiol.2003.08.005] [PMID: 14580852]
[21]
Mrsulja, B.B.; Mrsulja, B.J.; Fujimoto, T.; Klatzo, I.; Spatz, M. Isolation of brain capillaries: A simplified technique. Brain Res., 1976, 110(2), 361-365.
[http://dx.doi.org/10.1016/0006-8993(76)90408-X] [PMID: 938949]
[http://dx.doi.org/10.1016/0006-8993(76)90408-X] [PMID: 938949]
[22]
Cioni, C.; Turlizzi, E.; Zanelli, U.; Oliveri, G.; Annunziata, P. Expression of tight junction and drug efflux transporter proteins in an in vitro model of human blood–brain barrier. Front. Psychiatry, 2012, 3, 47.
[http://dx.doi.org/10.3389/fpsyt.2012.00047] [PMID: 22593745]
[http://dx.doi.org/10.3389/fpsyt.2012.00047] [PMID: 22593745]
[23]
Tada, H.; Shiho, O.; Kuroshima, K.; Koyama, M.; Tsukamoto, K. An improved colorimetric assay for interleukin 2. J. Immunol. Methods, 1986, 93(2), 157-165.
[http://dx.doi.org/10.1016/0022-1759(86)90183-3] [PMID: 3490518]
[http://dx.doi.org/10.1016/0022-1759(86)90183-3] [PMID: 3490518]
[24]
van der Zee, J.M.; Siegert, C.E.H.; de Vreede, T.A.; Daha, M.R.; Breedveld, F.C. Characterization of anti-endothelial cell antibodies in Systemic Lupus Erythematosus (SLE). Clin. Exp. Immunol., 1991, 84(2), 238-244.
[PMID: 2025951]
[PMID: 2025951]
[25]
Schägger, H.; von Jagow, G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal. Biochem., 1987, 166(2), 368-379.
[http://dx.doi.org/10.1016/0003-2697(87)90587-2] [PMID: 2449095]
[http://dx.doi.org/10.1016/0003-2697(87)90587-2] [PMID: 2449095]
[26]
Galve-Roperh, I.; Sánchez, C.; Cortés, M.L.; Gómez del Pulgar, T.; Izquierdo, M.; Guzmán, M. Anti-tumoral action of cannabinoids: Involvement of sustained ceramide accumulation and extracellular signal-regulated kinase activation. Nat. Med., 2000, 6(3), 313-319.
[http://dx.doi.org/10.1038/73171] [PMID: 10700234]
[http://dx.doi.org/10.1038/73171] [PMID: 10700234]
[27]
Sánchez, C.; Galve-Roperh, I.; Canova, C.; Brachet, P.; Guzmán, M. Delta9-tetrahydrocannabinol induces apoptosis in C6 glioma cells. FEBS Lett., 1998, 436(1), 6-10.
[http://dx.doi.org/10.1016/S0014-5793(98)01085-0] [PMID: 9771884]
[http://dx.doi.org/10.1016/S0014-5793(98)01085-0] [PMID: 9771884]
[28]
Guzmán, M.; Sánchez, C.; Galve-Roperh, I. Control of the cell survival/death decision by cannabinoids. J. Mol. Med. (Berl.), 2001, 78(11), 613-625.
[http://dx.doi.org/10.1007/s001090000177] [PMID: 11269508]
[http://dx.doi.org/10.1007/s001090000177] [PMID: 11269508]
[29]
Stella, N. Cannabinoid signaling in glial cells. Glia, 2004, 48(4), 267-277.
[http://dx.doi.org/10.1002/glia.20084] [PMID: 15390110]
[http://dx.doi.org/10.1002/glia.20084] [PMID: 15390110]
[30]
Schley, M.; Ständer, S.; Kerner, J.; Vajkoczy, P.; Schüpfer, G.; Dusch, M.; Schmelz, M.; Konrad, C. Predominant CB2 receptor expression in endothelial cells of glioblastoma in humans. Brain Res. Bull., 2009, 79(5), 333-337.
[http://dx.doi.org/10.1016/j.brainresbull.2009.01.011] [PMID: 19480992]
[http://dx.doi.org/10.1016/j.brainresbull.2009.01.011] [PMID: 19480992]
[31]
Fernández-Ruiz, J.; Romero, J.; Velasco, G.; Tolón, R.M.; Ramos, J.A.; Guzmán, M. Cannabinoid CB2 receptor: A new target for controlling neural cell survival? Trends Pharmacol. Sci., 2007, 28(1), 39-45.
[http://dx.doi.org/10.1016/j.tips.2006.11.001] [PMID: 17141334]
[http://dx.doi.org/10.1016/j.tips.2006.11.001] [PMID: 17141334]
[32]
Palazuelos, J.; Aguado, T.; Egia, A.; Mechoulam, R.; Guzmán, M.; Galve-Roperh, I. Non-psychoactive CB2 cannabinoid agonists stimulate neural progenitor proliferation. FASEB J., 2006, 20(13), 2405-2407.
[http://dx.doi.org/10.1096/fj.06-6164fje] [PMID: 17015409]
[http://dx.doi.org/10.1096/fj.06-6164fje] [PMID: 17015409]
[33]
Guzmán, M.; Galve-Roperh, I.; Sánchez, C. Ceramide: A new second messenger of cannabinoid action. Trends Pharmacol. Sci., 2001, 22(1), 19-22.
[http://dx.doi.org/10.1016/S0165-6147(00)01586-8] [PMID: 11165667]
[http://dx.doi.org/10.1016/S0165-6147(00)01586-8] [PMID: 11165667]
[34]
Carracedo, A.; Lorente, M.; Egia, A.; Blázquez, C.; García, S.; Giroux, V.; Malicet, C.; Villuendas, R.; Gironella, M.; González-Feria, L.; Piris, M.A.; Iovanna, J.L.; Guzmán, M.; Velasco, G. The stress-regulated protein p8 mediates cannabinoid-induced apoptosis of tumor cells. Cancer Cell, 2006, 9(4), 301-312.
[http://dx.doi.org/10.1016/j.ccr.2006.03.005] [PMID: 16616335]
[http://dx.doi.org/10.1016/j.ccr.2006.03.005] [PMID: 16616335]
[35]
Massi, P.; Valenti, M.; Vaccani, A.; Gasperi, V.; Perletti, G.; Marras, E.; Fezza, F.; Maccarrone, M.; Parolaro, D. 5-Lipoxygenase and anandamide hydrolase (FAAH) mediate the antitumor activity of cannabidiol, a non-psychoactive cannabinoid. J. Neurochem., 2008, 104(4), 1091-1100.
[http://dx.doi.org/10.1111/j.1471-4159.2007.05073.x] [PMID: 18028339]
[http://dx.doi.org/10.1111/j.1471-4159.2007.05073.x] [PMID: 18028339]
[36]
Chang, H.L.; Gillett, N.; Figari, I.; Lopez, A.R.; Palladino, M.A.; Derynck, R. Increased transforming growth factor β expression inhibits cell proliferation in vitro, yet increases tumorigenicity and tumor growth of Meth A sarcoma cells. Cancer Res., 1993, 53(18), 4391-4398.
[PMID: 8364935]
[PMID: 8364935]
[37]
Cui, W.; Fowlis, D.J.; Bryson, S.; Duffie, E.; Ireland, H.; Balmain, A.; Akhurst, R.J. TGFbeta1 inhibits the formation of benign skin tumors, but enhances progression to invasive spindle carcinomas in transgenic mice. Cell, 1996, 86(4), 531-542.
[http://dx.doi.org/10.1016/S0092-8674(00)80127-0] [PMID: 8752208]
[http://dx.doi.org/10.1016/S0092-8674(00)80127-0] [PMID: 8752208]
[38]
Constam, D.B.; Philipp, J.; Malipiero, U.V.; ten Dijke, P.; Schachner, M.; Fontana, A. Differential expression of transforming growth factor-β 1, -β 2, and -β 3 by glioblastoma cells, astrocytes, and microglia. J. Immunol., 1992, 148(5), 1404-1410.
[PMID: 1538124]
[PMID: 1538124]
[39]
Kjellman, C.; Olofsson, S.P.; Hansson, O.; Von Schantz, T.; Lindvall, M.; Nilsson, I.; Salford, L.G.; Sjögren, H.O.; Widegren, B. Expression of TGF-β isoforms, TGF-β receptors, and SMAD molecules at different stages of human glioma. Int. J. Cancer, 2000, 89(3), 251-258.
[http://dx.doi.org/10.1002/1097-0215(20000520)89:3<251:AID-IJC7>3.0.CO;2-5] [PMID: 10861501]
[http://dx.doi.org/10.1002/1097-0215(20000520)89:3<251:AID-IJC7>3.0.CO;2-5] [PMID: 10861501]
[40]
Schneider, T.; Sailer, M.; Ansorge, S.; Firsching, R.; Reinhold, D. Increased concentrations of transforming growth factor beta1 and beta2 in the plasma of patients with glioblastoma. J. Neurooncol., 2006, 79(1), 61-65.
[http://dx.doi.org/10.1007/s11060-005-9116-7] [PMID: 16614941]
[http://dx.doi.org/10.1007/s11060-005-9116-7] [PMID: 16614941]
[41]
Pan, J.J.; Chang, W.J.; Barone, T.A.; Plunkett, R.J.; Ostrow, P.T.; Greenberg, S.J. Increased expression of TGF-β1 reduces tumor growth of human U-87 glioblastoma cells in vivo. Cancer Immunol. Immunother., 2006, 55(8), 918-927.
[http://dx.doi.org/10.1007/s00262-005-0083-9] [PMID: 16187082]
[http://dx.doi.org/10.1007/s00262-005-0083-9] [PMID: 16187082]
[42]
Ikushima, H.; Todo, T.; Ino, Y.; Takahashi, M.; Miyazawa, K.; Miyazono, K. Autocrine TGF-beta signaling maintains tumorigenicity of glioma-initiating cells through Sry-related HMG-box factors. Cell Stem Cell, 2009, 5(5), 504-514.
[http://dx.doi.org/10.1016/j.stem.2009.08.018] [PMID: 19896441]
[http://dx.doi.org/10.1016/j.stem.2009.08.018] [PMID: 19896441]