The Effect of the Synthetic Cannabinoid AB-CHMINACA on the Roles of Vascular Endothelial Growth Factor, Angiopoietin-1, and Angiopoietin-2 in Brain Angiogenesis
Abstract
Introduction:
Synthetic cannabinoids (SCs) are new psychoactive compound that mimic the actions of tetrahydrocannabinol, the main psychoactive ingredient in cannabis. Angiogenesis is the formation of new blood vessels from previously existing vessels, which is crucial in a variety of physiological and pathological circumstances, including drug addiction.
Objective:
The objective of this study was to investigate the influence of a specific SC compound (AB-CHMINACA) on angiogenesis and the gene expression of proangiogenic stimulators in human brain microvascular endothelial cells (HBMVECs).
Methods:
HBMVECs were cultured in 500 mL of DMEM media supplemented with specific additives as per the instructions provided in the endothelial cell growth kit. Cell viability was assessed using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) assay. The migration rates of HBMVECs were evaluated using the wound healing assay. In vitro tube formation was performed to assess the angiogenic ability of endothelial cells. The expression levels of glycogen synthase kinase 3 beta and various pro-angiogenic stimulators were quantified using western blot, Enzyme-Linked Immunosorbent Assay (ELISA), and quantitative polymerase chain reaction techniques.
Results:
AB-CHMINACA enhanced angiogenic activities, including endothelial cell migration, proliferation, and capillary tube-like formation in HBMVECs. Furthermore, it stimulated the expression of proangiogenic stimulators at both the RNA and protein levels.
Conclusion:
Our findings indicate that AB-CHMINACA promotes angiogenic processes in the human brain, including endothelial cell proliferation, migration, and tube formation. Additionally, it upregulates the expression of proangiogenic stimulators at the molecular level. These results suggest that AB-CHMINACA could be considered an inducer of brain angiogenesis and may contribute to the development of innovative treatment strategies for disorders associated with abnormal angiogenesis.
Get full access to this article
View all access and purchase options for this article.
References
1. Le Boisselier R, Alexandre J, Lelong-Boulouard V, et al. Focus on cannabinoids and synthetic cannabinoids. Clin Pharmacol Ther, 2017; 101(2):220–229.
2. Auwärter V, Dresen S, Weinmann W, et al. “Spice” and other herbal blends: Harmless incense or cannabinoid designer drugs?. J Mass Spectrom, 2009; 44(5):832–837.
3. Kikura-Hanajiri R, Uchiyama N, Goda Y. Survey of current trends in the abuse of psychotropic substances and plants in Japan. Legal Med, 2011; 13(3):109–115.
4. Farhoudian A, Radfar SR, Mohaddes Ardabili H, et al. A global survey on changes in the supply, price, and use of illicit drugs and alcohol, and related complications during the 2020 COVID-19 pandemic. Front Psychiatry, 2021; 12:646206.
5. Adamowicz P, Tokarczyk B. Simple and rapid screening procedure for 143 new psychoactive substances by liquid chromatography-tandem mass spectrometry. Drug Test Anal, 2016; 8(7):652–667.
6. Seely KA, Lapoint J, Moran JH, et al. Spice drugs are more than harmless herbal blends: A review of the pharmacology and toxicology of synthetic cannabinoids. Prog Neuropsychopharmacol Biol Psychiatry, 2012; 39(2):234–243.
7. Weaver MF, Hopper JA, Gunderson EW. Designer drugs 2015: Assessment and management. Addict Sci Clin Pract, 2015; 10(1):1–9.
8. Hermanns-Clausen M, Kithinji J, Spehl M, et al. Adverse effects after the use of JWH-210—A case series from the EU Spice II plus project. Drug Test Anal, 2016; 8(10):1030–1038.
9. Howlett AC, Barth F, Bonner T, et al. International Union of Pharmacology. XXVII. Classification of cannabinoid receptors. Pharmacol Rev, 2002; 54(2):161–202.
10. Pertwee RG. Pharmacological actions of cannabinoids. Springer: Berlin Heidelberg; 2005.
11. Antonides LH, Cannaert A, Norman C, et al. Enantiospecific synthesis, chiral separation, and biological activity of four indazole-3-carboxamide-type synthetic cannabinoid receptor agonists and their detection in seized drug samples. Front Chem, 2019; 7:321.
12. Uchiyama N, Matsuda S, Wakana D, et al. New cannabimimetic indazole derivatives, N-(1-amino-3-methyl-1-oxobutan-2-yl)-1-pentyl-1 H-indazole-3-carboxamide (AB-PINACA) and N-(1-amino-3-methyl-1-oxobutan-2-yl)-1-(4-fluorobenzyl)-1 H-indazole-3-carboxamide (AB-FUBINACA) identified as designer drugs in illegal products. Forensic Toxicol, 2013; 31:93–100.
13. Sim J, Cho HS, Lee J, et al. Determination of AB-CHMINACA and its metabolites in human hair and their deposition in hair of abusers. J Pharm Biomed Anal, 2017; 140:162–168.
14. Wiley JL, Marusich JA, Lefever TW, et al. AB-CHMINACA, AB-PINACA, and FUBIMINA: Affinity and potency of novel synthetic cannabinoids in producing Δ9-Tetrahydrocannabinol–like effects in mice. J Pharmacol Exp Ther, 2015; 354(3):328–339.
15. Tyndall JA, Gerona R, De Portu G, et al. An outbreak of acute delirium from exposure to the synthetic cannabinoid AB-CHMINACA. Clin Toxicol, 2015; 53(10):950–956.
16. Folkman J. Clinical applications of research on angiogenesis. N Engl J Med, 1995; 333(26):1757–1763.
17. Moses KM, Klagsbrun M. Molecular angiogenesis. Chem Biol, 1999; 6:R217–R224.
18. Eelen G, Treps L, Li X, et al. Basic and therapeutic aspects of angiogenesis updated. Circ Res, 2020; 127(2):310–329
19. Al-Eitan L, Alhusban A, Alahmad S. Effects of the synthetic cannabinoid XLR-11 on the viability and migration rates of human brain microvascular endothelial cells in a clinically-relevant model. Pharmacol Rep, 2020; 72:1717–1724.
20. Folkman J. Angiogenesis in cancer, vascular, rheumatoid and other disease. Nat Med, 1995; 1(1):27–30.
21. Pluda JM. Tumor-associated angiogenesis: Mechanisms, clinical implications, and therapeutic strategies. Seminars Oncol, 1997; 24(2):203–218.
22. Colpaert C, Vermeulen P, Benoy I, et al. Inflammatory breast cancer shows angiogenesis with high endothelial proliferation rate and strong E-cadherin expression. Br J Cancer, 2003; 88(5):718–725.
23. Xiang Y, Yao X, Wang X, et al. Houshiheisan promotes angiogenesis via HIF-1α/VEGF and SDF-1/CXCR4 pathways: In vivo and in vitro. Biosci Rep, 2019; 39(10):BSR20191006.
24. Erratico C, Negreira N, Norouzizadeh H, et al. In vitro and in vivo human metabolism of the synthetic cannabinoid AB-CHMINACA. Drug Test Anal, 2015; 7(10):866–876.
25. Hasegawa K, Wurita A, Minakata K, et al. Postmortem distribution of MAB-CHMINACA in body fluids and solid tissues of a human cadaver. Forensic Toxicol, 2015; 33:380–387.
26. Institóris L, Hidvégi E, Kovács K, et al. Drug consumption of suspected drug-influenced drivers in Hungary (2016–2018). Forensic Sci Int, 2022; 336:111325.
27. Wiley JL, Martin BR. Preclinical Pharmacological and Brain Bioassay Systems for CB1 Cannabinoid Receptors, in the Cannabinoid Receptors (Reggio PH. ed.). Humana Press: Totowa, NJ; 2009; pp. 329–360.
28. Wiley JL, Compton DR, Dai D, et al. Structure-activity relationships of indole-and pyrrole-derived cannabinoids. J Pharmacol Exp Ther, 1998; 285(3):995–1004.
29. Compton DR, Gold LH, Ward SJ, et al. Aminoalkylindole analogs: Cannabimimetic activity of a class of compounds structurally distinct from delta 9-tetrahydrocannabinol. J Pharmacol Exp Ther, 1992; 263(3):1118–1126.
30. Almada M, Alves P, Fonseca BM, et al. Synthetic cannabinoids JWH-018, JWH-122, UR-144 and the phytocannabinoid THC activate apoptosis in placental cells. Toxicol Lett, 2020; 319:129–137;
31. Couceiro J, Bandarra S, Sultan H, et al. Toxicological impact of JWH-018 and its phase I metabolite N-(3-hydroxypentyl) on human cell lines. Forensic Sci Int, 2016; 264:100–105;
32. Jin K, Xie L, Kim SH, et al. Defective adult neurogenesis in CB1 cannabinoid receptor knockout mice. Mol Pharmacol, 2004; 66(2):204–208;
33. Aguado T, Romero E, Monory K, et al. The CB1 cannabinoid receptor mediates excitotoxicity-induced neural progenitor proliferation and neurogenesis. J Biol Chem, 2007; 282(33):23892–23898;
34. Grimaldi C, Pisanti S, Laezza C, et al. Anandamide inhibits adhesion and migration of breast cancer cells. Exp Cell Res, 2006; 312(4):363–373.
35. Nabissi M, Morelli MB, Offidani M, et al. Cannabinoids synergize with carfilzomib, reducing multiple myeloma cells viability and migration. Oncotarget, 2016; 7(47):77543.
36. Preet A, Ganju R, Groopman J. Δ9-Tetrahydrocannabinol inhibits epithelial growth factor-induced lung cancer cell migration in vitro as well as its growth and metastasis in vivo. Oncogene, 2008; 27(3):339–346.
37. Liu C, Qi X, Alhabeil J, et al. Activation of cannabinoid receptors promote periodontal cell adhesion and migration. J Clin Periodontol, 2019; 46(12):1264–1272;
38. Luo H, Rossi E, Saubamea B, et al. Cannabidiol increases proliferation, migration, tubulogenesis, and integrity of human brain endothelial cells through TRPV2 activation. Mol Pharm, 2019; 16(3):1312–1326;
39. Vendel E, de Lange E. Functions of the CB1 and CB2 receptors in neuroprotection at the level of the blood–brain barrier. Neuromol Med, 2014; 16(3):620–642.
40. Daly C, Eichten A, Castanaro C, et al. Angiopoietin-2 functions as a Tie2 agonist in tumor models, where it limits the effects of VEGF inhibition. Cancer Res, 2013; 73(1):108–118.
41. Saharinen P, Eklund L, Alitalo K. Therapeutic targeting of the angiopoietin–TIE pathway. Nat Rev Drug Discov, 2017; 16(9):635–661.
42. Augustin HG, Young Koh G, Thurston G, et al. Control of vascular morphogenesis and homeostasis through the angiopoietin–Tie system. Nat Rev Mol Cell Biol, 2009; 10(3):165–177.
43. Huang H, Bhat A, Woodnutt G, et al. Targeting the ANGPT–TIE2 pathway in malignancy. Nat Rev Cancer, 2010; 10(8):575–585.
44. Melincovici CS, Boşca AB, Şuşman S, et al. Vascular endothelial growth factor (VEGF)-key factor in normal and pathological angiogenesis. Rom J Morphol Embryol, 2018; 59(2):455–467.
45. Neufeld G, Cohen T, Gengrinovitch S, et al. Vascular endothelial growth factor (VEGF) and its receptors. FASEB J, 1999; 13(1):9–22.
46. Arnaoutova I, Kleinman HK. In vitro angiogenesis: Endothelial cell tube formation on gelled basement membrane extract. Nat Protoc, 2010; 5(4):628–635.
47. Brunton VG, Frame MC. Src and focal adhesion kinase as therapeutic targets in cancer. Curr Opin Pharmacol, 2008; 8(4):427–432.
48. Thapa D, Lee JS, Heo SW, et al. Novel hexahydrocannabinol analogs as potential anti-cancer agents inhibit cell proliferation and tumor angiogenesis. Eur J Pharmacol, 2011; 650(1):64–71;
49. Zhang Z, Chopp M. Vascular endothelial growth factor and angiopoietins in focal cerebral ischemia. Trends Cardiovasc Med, 2002; 12(2):62–66.
50. Carmeliet P. Mechanisms of angiogenesis and arteriogenesis. Nat Med, 2000; 6(4):389–395.
51. Casanova ML, Larcher F, Casanova B, et al. A critical role for ras-mediated, epidermal growth factor receptor-dependent angiogenesis in mouse skin carcinogenesis. Cancer Res, 2002; 62(12):3402–3407.
52. Holash J, Wiegand S, Yancopoulos G. New model of tumor angiogenesis: Dynamic balance between vessel regression and growth mediated by angiopoietins and VEGF. Oncogene, 1999; 18(38):5356–5362.
53. Maher EA, Furnari FB, Bachoo RM, et al. Malignant glioma: Genetics and biology of a grave matter. Genes Dev, 2001; 15(11):1311–1333.
54. Blázquez C, Casanova ML, Planas A, et al. Inhibition of tumor angiogenesis by cannabinoids. FASEB J, 2003; 17(3):1–16.
55. Casanova ML, Blázquez C, Martínez-Palacio J, et al. Inhibition of skin tumor growth and angiogenesis in vivo by activation of cannabinoid receptors. J Clin Invest, 2003; 111(1):43–50.
56. Embi N, Rylatt DB, Cohen P. Glycogen synthase kinase-3 from rabbit skeletal muscle: Separation from cyclic-AMP-dependent protein kinase and phosphorylase kinase. Eur J Biochem, 1980; 107(2):519–527
57. Forde Ja, Dale TC. Glycogen synthase kinase 3: A key regulator of cellular fate. Cell Mol Life Sci, 2007; 64:1930–1944.
58. Cormier KW, Woodgett JR. Recent advances in understanding the cellular roles of GSK-3. F1000Res, 2017; 6:F1000 Faculty Rev-167;
59. Doble BW, Woodgett JR. GSK-3: Tricks of the trade for a multi-tasking kinase. J Cell Sci, 2003; 116(7):1175–1186.
60. Grimes CA, Jope RS. The multifaceted roles of glycogen synthase kinase 3β in cellular signaling. Progr Neurobiol, 2001; 65(4):391–426.
61. Frame S, Cohen P. GSK3 takes centre stage more than 20 years after its discovery. Biochem J, 2001; 359(1):1–16.
62. Woodgett JR. Molecular cloning and expression of glycogen synthase kinase-3/factor A. EMBO J, 1990; 9(8):2431–2438.
63. Ryves WJ, Dajani R, Pearl L, et al. Glycogen synthase kinase-3 inhibition by lithium and beryllium suggests the presence of two magnesium binding sites. Biochem Biophys Res Commun, 2002; 290(3):967–972.
64. Beurel E, Grieco SF, Jope RS. Glycogen synthase kinase-3 (GSK3): Regulation, actions, and diseases. Pharmacol Ther, 2015; 148:114–131.
65. Cervello M, Augello G, Cusimano A, et al. Pivotal roles of glycogen synthase-3 in hepatocellular carcinoma. Adv Biol Regul, 2017; 65:59–76.
66. Domoto T, Pyko IV, Furuta T, et al. Glycogen synthase kinase-3β is a pivotal mediator of cancer invasion and resistance to therapy. Cancer Sci, 2016; 107(10):1363–1372.
67. McCubrey JA, Rakus D, Gizak A, et al. Effects of mutations in Wnt/β-catenin, hedgehog, Notch and PI3K pathways on GSK-3 activity—Diverse effects on cell growth, metabolism and cancer. Biochim Biophys Acta, 2016; 1863(12):2942–2976.
68. McCubrey JA, Steelman LS, Bertrand FE, et al. Multifaceted roles of GSK-3 and Wnt/β-catenin in hematopoiesis and leukemogenesis: Opportunities for therapeutic intervention. Leukemia, 2014; 28(1):15–33.
69. Williams SP, Nowicki MO, Liu F, et al. Indirubins decrease glioma invasion by blocking migratory phenotypes in both the tumor and stromal endothelial cell compartments. Cancer Res, 2011; 71(16):5374–5380.
70. Wang L, Li J, Di Lj. Glycogen synthesis and beyond, a comprehensive review of GSK3 as a key regulator of metabolic pathways and a therapeutic target for treating metabolic diseases. Med Res Rev, 2022; 42(2):946–982.
71. Ma R, Xie Q, Li H, et al. l-Borneol exerted the neuroprotective effect by promoting angiogenesis coupled with neurogenesis via Ang1-VEGF-BDNF pathway. Front Pharmacol, 2021; 12:641894.
72. Jope RS, Johnson GV. The glamour and gloom of glycogen synthase kinase-3. Trends Biochem Sci, 2004; 29(2):95–102;
73. Ozaita A, Puighermanal E, Maldonado R. Regulation of PI3K/Akt/GSK-3 pathway by cannabinoids in the brain. J Neurochem, 2007; 102(4):1105–1114;
74. Mai W, Kawakami K, Shakoori A, et al. Deregulated GSK3{beta} sustains gastrointestinal cancer cells survival by modulating human telomerase reverse transcriptase and telomerase. Clin Cancer Res, 2009; 15(22):6810–6819;
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
Cite
Cite
Cite
OR
Download to reference manager
If you have citation software installed, you can download citation data to the citation manager of your choice
Information, rights and permissions
Information
Published In
Article first published online: September 19, 2023
Issue published: September 1, 2023
Keywords
Authors
Authors' Contributions
The research was designed by L.A.-E. L.A.-E., and H.A.K. carried out the experimental work. The data were statistically analyzed and interpreted by L.A.-E. and H.A.K. The manuscript was written by L.A.-E. and H.A.K. All authors contributed to the manuscript review.
Metrics and citations
Metrics
Journals metrics
This article was published in Applied In Vitro Toxicology.
View All Journal MetricsPublication usage*
Total views and downloads: 58
*Publication usage tracking started in December 2016
Publications citing this one
Receive email alerts when this publication is cited
Web of Science: 0
Crossref: 5
- The Impact of Potent Addictive Substances on Angiogenic Behavior: A Comprehensive Review
- Evaluation of the metabolic activity, angiogenic impacts, and GSK-3β signaling of the synthetic cannabinoid MMB-2201 on human cerebral microvascular endothelial cells
- RETRATCED ARTICLE: Effect of EMB-FUBINACA on brain endothelial cell angiogenesis: Expression analysis of angiogenic markers
- The synthetic cannabinoid 5-fluoro ABICA upregulates angiogenic markers and stimulates tube formation in human brain microvascular endothelial cells
- MDMB-FUBINACA Influences Brain Angiogenesis and the Expression of VEGF, ANG-1, and ANG-2
Figures and tables
Figures & Media
Tables
View Options
Access options
If you have access to journal content via a personal subscription, university, library, employer or society, select from the options below:
I am signed in as:
View my profileSign out
I can access personal subscriptions, purchases, paired institutional access and free tools such as favourite journals, email alerts and saved searches.
loading institutional access options
Alternatively, view purchase options below:
Purchase 24 hour online access to view and download content.
Access journal content via a DeepDyve subscription or find out more about this option.
