Abstract
Cannabinoid signaling, mainly via CB1 and CB2 receptors, plays an essential role in oligodendrocyte health and functions. However, the specific molecular signals associated with the activation or blockade of CB1 and CB2 receptors in this glial cell have yet to be elucidated. Mass spectrometry-based shotgun proteomics and in silico biology tools were used to determine which signaling pathways and molecular mechanisms are triggered in a human oligodendrocytic cell line (MO3.13) by several pharmacological stimuli: the phytocannabinoid cannabidiol (CBD); CB1 and CB2 agonists ACEA, HU308, and WIN55, 212–2; CB1 and CB2 antagonists AM251 and AM630; and endocannabinoids anandamide (AEA) and 2-arachidonoylglycerol (2-AG). The modulation of cannabinoid signaling in MO3.13 was found to affect pathways linked to cell proliferation, migration, and differentiation of oligodendrocyte progenitor cells. Additionally, we found that carbohydrate and lipid metabolism, as well as mitochondrial function, were modulated by these compounds. Comparing the proteome changes and upstream regulators among treatments, the highest overlap was between the CB1 and CB2 antagonists, followed by overlaps between AEA and 2-AG. Our study opens new windows of opportunities, suggesting that cannabinoid signaling in oligodendrocytes might be relevant in the context of demyelinating and neurodegenerative diseases. Proteomics data are available at ProteomeXchange (PXD031923).
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Data availability
The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE (PubMed ID: 34,723,319) partner repository with the dataset identifier PXD031923 [90].
Abbreviations
- 2-AG:
-
2-Arachidonoylglycerol
- ACN:
-
Acetonitrile
- AEA:
-
Anandamide
- BSA:
-
Bovine serum albumin
- CB1:
-
Cannabinoid receptor, type 1
- CB2:
-
Cannabinoid receptor, type 2
- CBD:
-
Cannabidiol
- CNS:
-
Central nervous system
- D2:
-
Dopamine receptor, type 2
- DAGLA:
-
Diacylglycerol lipase alpha
- DAGLB:
-
Diacylglycerol lipase beta
- DEPs:
-
Differentially expressed proteins
- DMEM:
-
Dulbecco’s modified Eagle medium
- ECBs:
-
Endocannabinoids
- EIF2:
-
Eukaryotic initiation factor 2
- ESI:
-
Electrospray ionization
- FAAH:
-
Fatty acid amide hydrolase
- FDR:
-
False discovery rate
- HBSS:
-
Hank’s balanced salt solution
- ICC:
-
Immunocytochemistry
- IPA:
-
Ingenuity pathway analysis
- LC:
-
Liquid chromatography
- MAGL:
-
Monoacylglycerol lipase
- MAPK:
-
Mitogen-activated protein kinase
- MBP:
-
Myelin binding protein
- MS:
-
Mass spectrometry
- MS-TOF:
-
Time-of-flight mass spectrometry
- NAPE-PLD:
-
N-acyl phosphatidylethanolamine phospholipase D
- OPC:
-
Oligodendrocyte precursor cell
- PBS:
-
Phosphate-buffered saline
- RT:
-
Room temperature
- TBS:
-
Tris-buffered saline
- THC:
-
Tetrahydrocannabinol
- TRPV1:
-
Transient receptor potential cation channel, subfamily V, member 1
- UPLC:
-
Ultra-performance liquid chromatography
- VWM:
-
Vanishing white matter
References
Howlett AC (1985) Cannabinoid inhibition of adenylate cyclase. Biochemistry of the response in neuroblastoma cell membranes. Mol Pharmacol 27:429–436
Matsuda LA, Lolait SJ, Brownstein MJ et al (1990) Structure of a cannabinoid receptor and functional expression of the cloned cDNA. Nature 346:561–564
Munro S, Thomas KL, Abu-Shaar M (1993) Molecular characterization of a peripheral receptor for cannabinoids. Nature 365:61–65
Devane WA, Dysarz FA 3rd, Johnson MR et al (1988) Determination and characterization of a cannabinoid receptor in rat brain. Mol Pharmacol 34:605–613
Di Marzo V, Piscitelli F (2015) The endocannabinoid system and its modulation by phytocannabinoids. Neurotherapeutics 12:692–698
Howlett AC (1998) The CB1 cannabinoid receptor in the brain. Neurobiol Dis 5:405–416
Howlett AC, Abood ME (2017) CB and CB Receptor Pharmacology. Adv Pharmacol 80:169–206
Van Sickle MD, Duncan M, Kingsley PJ et al (2005) Identification and functional characterization of brainstem cannabinoid CB2 receptors. Science 310:329–332
Lisboa SF, Gomes FV, Guimaraes FS, Campos AC (2016) Microglial Cells as a Link between Cannabinoids and the Immune Hypothesis of Psychiatric Disorders. Front Neurol 7:5
Rodrigues LCM, Gobira PH, de Oliveira AC et al (2014) Neuroinflammation as a possible link between cannabinoids and addiction. Acta Neuropsychiatr 26:334–346
Huerga-Gómez A, Aguado T, Sánchez-de la Torre A et al (2021) Δ -Tetrahydrocannabinol promotes oligodendrocyte development and CNS myelination in vivo. Glia 69:532–545
de Almeida V, Martins-de-Souza D (2018) Cannabinoids and glial cells: possible mechanism to understand schizophrenia. Eur Arch Psychiatry Clin Neurosci 268:727–737
Katona I, Sperlágh B, Sı́k A, et al (1999) Presynaptically located CB1 cannabinoid receptors regulate GABA release from axon terminals of specific hippocampal interneurons. J Neurosci 19:4544–4558
Stella N (2010) Cannabinoid and cannabinoid-like receptors in microglia, astrocytes, and astrocytomas. Glia 58:1017–1030
Fünfschilling U, Supplie LM, Mahad D et al (2012) Glycolytic oligodendrocytes maintain myelin and long-term axonal integrity. Nature 485:517–521
Lee Y, Morrison BM, Li Y et al (2012) Oligodendroglia metabolically support axons and contribute to neurodegeneration. Nature 487:443–448
Molina-Holgado E, Vela JM, Arévalo-Martín A et al (2002) Cannabinoids promote oligodendrocyte progenitor survival: involvement of cannabinoid receptors and phosphatidylinositol-3 kinase/Akt signaling. J Neurosci 22:9742–9753
Solbrig MV, Fan Y, Hermanowicz N et al (2010) A synthetic cannabinoid agonist promotes oligodendrogliogenesis during viral encephalitis in rats. Exp Neurol 226:231–241
Gomez O, Sanchez-Rodriguez A, Le M et al (2011) Cannabinoid receptor agonists modulate oligodendrocyte differentiation by activating PI3K/Akt and the mammalian target of rapamycin (mTOR) pathways. Br J Pharmacol 163:1520–1532
Tomas-Roig J, Wirths O, Salinas-Riester G, Havemann-Reinecke U (2016) The cannabinoid CB1/CB2 agonist WIN55212.2 promotes oligodendrocyte differentiation in vitro and neuroprotection during the cuprizone-induced central nervous system demyelination. CNS Neurosci Ther 22:387–395
Moreno-Luna R, Esteban PF, Paniagua-Torija B et al (2021) Heterogeneity of the endocannabinoid system between cerebral cortex and spinal cord oligodendrocytes. Mol Neurobiol 58:689–702
Ilyasov AA, Milligan CE, Pharr EP, Howlett AC (2018) The Endocannabinoid system and oligodendrocytes in health and disease. Front Neurosci 12:733
Mecha M, Carrillo-Salinas FJ, Feliú A et al (2020) Perspectives on cannabis-based therapy of multiple sclerosis: a mini-review. Front Cell Neurosci 14:34
Jinsmaa Y, Isonaka R, Sharabi Y, Goldstein DS (2020) 3,4-Dihydroxyphenylacetaldehyde is more efficient than dopamine in oligomerizing and quinonizing -synuclein. J Pharmacol Exp Ther 372:157–165
Iwata K, Café-Mendes CC, Schmitt A et al (2013) The human oligodendrocyte proteome. Proteomics 13:3548–3553
Buntinx M, Vanderlocht J, Hellings N et al (2003) Characterization of three human oligodendroglial cell lines as a model to study oligodendrocyte injury: morphology and oligodendrocyte-specific gene expression. J Neurocytol 32:25–38
Brandão-Teles C, de Almeida V, Cassoli JS, Martins-de-Souza D (2019) Biochemical pathways triggered by antipsychotics in human [corrected] oligodendrocytes: potential of discovering new treatment targets. Front Pharmacol 10:186
Seabra G, de Almeida V, Reis-de-Oliveira G et al (2020) Ubiquitin-proteasome system, lipid metabolism and DNA damage repair are triggered by antipsychotic medication in human oligodendrocytes: implications in schizophrenia. Sci Rep 10:12655
Cassoli JS, Brandão-Teles C, Santana AG et al (2018) Ion mobility-enhanced data-independent acquisitions enable a deep proteomic landscape of oligodendrocytes. Proteomics. https://doi.org/10.1002/pmic.2018700015
Silva JC, Denny R, Dorschel CA et al (2005) Quantitative proteomic analysis by accurate mass retention time pairs. Anal Chem 77:2187–2200
Snel B, Lehmann G, Bork P, Huynen MA (2000) STRING: a web-server to retrieve and display the repeatedly occurring neighbourhood of a gene. Nucleic Acids Res 28:3442–3444
Fabregat A, Sidiropoulos K, Garapati P et al (2016) The reactome pathway knowledgebase. Nucleic Acids Res 44:D481–D487
Uhlen M, Ponten F, Lindskog C (2015) Charting the human proteome: Understanding disease using a tissue-based atlas. Science 347:1274–1274
Yu G, Wang L-G, Han Y, He Q-Y (2012) clusterProfiler: an R package for comparing biological themes among gene clusters. OMICS 16:284–287
Computing RFS (2013) R: A language and environment for statistical computing. R Core Team, Vienna
Tatomir A, Rao G, Boodhoo D et al (2020) Histone deacetylase SIRT1 mediates C5b-9-induced cell cycle in oligodendrocytes. Front Immunol 11:619
Prozorovski T, Ingwersen J, Lukas D et al (2019) Regulation of sirtuin expression in autoimmune neuroinflammation: Induction of SIRT1 in oligodendrocyte progenitor cells. Neurosci Lett 704:116–125
Hiratsuka D, Kurganov E, Furube E et al (2019) VEGF- and PDGF-dependent proliferation of oligodendrocyte progenitor cells in the medulla oblongata after LPC-induced focal demyelination. J Neuroimmunol 332:176–186
Mindos T, Dun X-P, North K et al (2017) Merlin controls the repair capacity of Schwann cells after injury by regulating Hippo/YAP activity. J Cell Biol 216:495–510
Azevedo MM, Domingues HS, Cordelières FP et al (2018) Jmy regulates oligodendrocyte differentiation via modulation of actin cytoskeleton dynamics. Glia 66:1826–1844
Thomason EJ, Escalante M, Osterhout DJ, Fuss B (2019) The oligodendrocyte growth cone and its actin cytoskeleton: A fundamental element for progenitor cell migration and CNS myelination. Glia. https://doi.org/10.1002/glia.23735
Nicaise AM, Johnson KM, Willis CM et al (2019) TIMP-1 promotes oligodendrocyte differentiation through receptor-mediated signaling. Mol Neurobiol 56:3380–3392
Harlow DE, Saul KE, Komuro H, Macklin WB (2015) Myelin proteolipid protein complexes with αv integrin and AMPA receptors In vivo and regulates ampa-dependent oligodendrocyte progenitor cell migration through the modulation of cell-surface GluR2 expression. J Neurosci 35:12018–12032
Linneberg C, Harboe M, Laursen LS (2015) Axo-glia interaction preceding CNS myelination is regulated by bidirectional eph-ephrin signaling. ASN Neuro. https://doi.org/10.1177/1759091415602859
Câmara J, Wang Z, Nunes-Fonseca C et al (2009) Integrin-mediated axoglial interactions initiate myelination in the central nervous system. J Cell Biol 185:699–712
Jagielska A, Lowe AL, Makhija E et al (2017) Mechanical strain promotes oligodendrocyte differentiation by global changes of gene expression. Front Cell Neurosci 11:93
Harboe M, Torvund-Jensen J, Kjaer-Sorensen K, Laursen LS (2018) Ephrin-A1-EphA4 signaling negatively regulates myelination in the central nervous system. Glia 66:934–950
Liang X, Draghi NA, Resh MD (2004) Signaling from integrins to Fyn to Rho family GTPases regulates morphologic differentiation of oligodendrocytes. J Neurosci 24:7140–7149
Ackerman SD, Garcia C, Piao X et al (2015) The adhesion GPCR Gpr56 regulates oligodendrocyte development via interactions with Gα12/13 and RhoA. Nat Commun 6:6122
Thurnherr T, Benninger Y, Wu X et al (2006) Cdc42 and Rac1 signaling are both required for and act synergistically in the correct formation of myelin sheaths in the CNS. J Neurosci 26:10110–10119
Seixas AI, Azevedo MM, Paes de Faria J et al (2019) Evolvability of the actin cytoskeleton in oligodendrocytes during central nervous system development and aging. Cell Mol Life Sci 76:1–11
Maki T, Choi YK, Miyamoto N et al (2018) A-kinase anchor protein 12 is required for oligodendrocyte differentiation in adult white matter. Stem Cells 36:751–760
Ghidinelli M, Poitelon Y, Shin YK et al (2017) Laminin 211 inhibits protein kinase A in Schwann cells to modulate neuregulin 1 type III-driven myelination. PLoS Biol 15:e2001408
Yang Y, Wang H, Zhang J et al (2013) Cyclin dependent kinase 5 is required for the normal development of oligodendrocytes and myelin formation. Dev Biol 378:94–106
Luo F, Burke K, Kantor C et al (2014) Cyclin-dependent kinase 5 mediates adult OPC maturation and myelin repair through modulation of Akt and GsK-3β signaling. J Neurosci 34:10415–10429
Miyamoto Y, Yamauchi J, Chan JR et al (2007) Cdk5 regulates differentiation of oligodendrocyte precursor cells through the direct phosphorylation of paxillin. J Cell Sci 120:4355–4366
Colognato H, Tzvetanova ID (2011) Glia unglued: how signals from the extracellular matrix regulate the development of myelinating glia. Dev Neurobiol 71:924–955
Hussain R, Macklin WB (2017) Integrin-linked kinase (ILK) deletion disrupts oligodendrocyte development by altering cell cycle. J Neurosci 37:397–412
Pronk JC, van Kollenburg B, Scheper GC, van der Knaap MS (2006) Vanishing white matter disease: a review with focus on its genetics. Ment Retard Dev Disabil Res Rev 12:123–128
Carter CJ (2007) eIF2B and oligodendrocyte survival: where nature and nurture meet in bipolar disorder and schizophrenia? Schizophr Bull 33:1343–1353
Arévalo-Martín A, García-Ovejero D, Rubio-Araiz A et al (2007) Cannabinoids modulate Olig2 and polysialylated neural cell adhesion molecule expression in the subventricular zone of post-natal rats through cannabinoid receptor 1 and cannabinoid receptor 2. Eur J Neurosci 26:1548–1559
Kuhn S, Gritti L, Crooks D, Dombrowski Y (2019) Oligodendrocytes in development, myelin generation and beyond. Cells. https://doi.org/10.3390/cells8111424
Gaesser JM, Fyffe-Maricich SL (2016) Intracellular signaling pathway regulation of myelination and remyelination in the CNS. Exp Neurol 283:501–511
Bernal-Chico A, Canedo M, Manterola A et al (2015) Blockade of monoacylglycerol lipase inhibits oligodendrocyte excitotoxicity and prevents demyelination in vivo. Glia 63:163–176
Sanchez-Rodriguez MA, Gomez O, Esteban PF et al (2018) The endocannabinoid 2-arachidonoylglycerol regulates oligodendrocyte progenitor cell migration. Biochem Pharmacol 157:180–188
Nave K-A, Werner HB (2014) Myelination of the nervous system: mechanisms and functions. Annu Rev Cell Dev Biol 30:503–533
Hussain G, Wang J, Rasul A et al (2019) Role of cholesterol and sphingolipids in brain development and neurological diseases. Lipids Health Dis 18:26
Voskuhl RR, Itoh N, Tassoni A et al (2019) Gene expression in oligodendrocytes during remyelination reveals cholesterol homeostasis as a therapeutic target in multiple sclerosis. Proc Natl Acad Sci U S A 116:10130–10139
Lin J-P, Mironova YA, Shrager P, Giger RJ (2017) LRP1 regulates peroxisome biogenesis and cholesterol homeostasis in oligodendrocytes and is required for proper CNS myelin development and repair. Elife. https://doi.org/10.7554/eLife.30498
Oddi S, Caporali P, Dragotto J et al (2019) The endocannabinoid system is affected by cholesterol dyshomeostasis: Insights from a murine model of Niemann Pick type C disease. Neurobiol Dis 130:104531
Suo N, Guo Y-E, He B et al (2019) Inhibition of MAPK/ERK pathway promotes oligodendrocytes generation and recovery of demyelinating diseases. Glia 67:1320–1332
Wong YL, LeBon L, Basso AM et al (2019) eIF2B activator prevents neurological defects caused by a chronic integrated stress response. Elife. https://doi.org/10.7554/eLife.42940
Melas PA, Qvist JS, Deidda M et al (2018) Cannabinoid modulation of eukaryotic initiation factors (eIF2α and eIF2B1) and behavioral cross-sensitization to cocaine in adolescent rats. Cell Rep 22:2909–2923
Terumitsu-Tsujita M, Kitaura H, Miura I et al (2019) Glial pathology in a novel spontaneous mutant mouse of the Eif2b5 gene: a vanishing white matter disease model. J Neurochem. https://doi.org/10.1111/jnc.14887
Vrechi TA, Crunfli F, Costa AP, Torrão AS (2018) Cannabinoid receptor Type 1 Agonist ACEA protects neurons from death and attenuates endoplasmic reticulum stress-related apoptotic pathway signaling. Neurotox Res 33:846–855
Harris JJ, Attwell D (2012) The energetics of CNS white matter. J Neurosci 32:356–371
Rinholm JE, Vervaeke K, Tadross MR et al (2016) Movement and structure of mitochondria in oligodendrocytes and their myelin sheaths. Glia 64:810–825
Rao VTS, Khan D, Cui Q-L et al (2017) Distinct age and differentiation-state dependent metabolic profiles of oligodendrocytes under optimal and stress conditions. PLoS ONE 12:e0182372
Scarante FF, Ribeiro MA, Almeida-Santos AF et al (2020) Glial cells and their contribution to the mechanisms of action of cannabidiol in neuropsychiatric disorders. Front Pharmacol 11:618065
Barley K, Dracheva S, Byne W (2009) Subcortical oligodendrocyte- and astrocyte-associated gene expression in subjects with schizophrenia, major depression and bipolar disorder. Schizophr Res 112:54–64
Morabito S, Miyoshi E, Michael N et al (2021) Single-nucleus chromatin accessibility and transcriptomic characterization of Alzheimer’s disease. Nat Genet 53:1143–1155
Jäkel S, Agirre E, Mendanha Falcão A et al (2019) Altered human oligodendrocyte heterogeneity in multiple sclerosis. Nature 566:543–547
Osorio-Querejeta I, Sáenz-Cuesta M, Muñoz-Culla M, Otaegui D (2017) Models for studying myelination, demyelination and remyelination. Neuromolecular Med 19:181–192
Campos AC, Ortega Z, Palazuelos J et al (2013) The anxiolytic effect of cannabidiol on chronically stressed mice depends on hippocampal neurogenesis: involvement of the endocannabinoid system. Int J Neuropsychopharmacol 16:1407–1419
Levin R, Peres FF, Almeida V et al (2014) Effects of cannabinoid drugs on the deficit of prepulse inhibition of startle in an animal model of schizophrenia: the SHR strain. Front Pharmacol 5:10
Almeida V, Peres FF, Levin R et al (2014) Effects of cannabinoid and vanilloid drugs on positive and negative-like symptoms on an animal model of schizophrenia: The SHR strain. Schizophr Res 153:150–159
Almeida V, Levin R, Peres FF et al (2013) Cannabidiol exhibits anxiolytic but not antipsychotic property evaluated in the social interaction test. Prog Neuropsychopharmacol Biol Psychiatry 41:30–35
Gomez O, Sanchez-Rodriguez MA, Ortega-Gutierrez S et al (2015) A basal tone of 2-arachidonoylglycerol contributes to early oligodendrocyte progenitor proliferation by activating phosphatidylinositol 3-kinase (PI3K)/AKT and the mammalian target of rapamycin (MTOR) pathways. J Neuroimmune Pharmacol 10:309–317
Almeida V, Levin R, Peres FF et al (2019) Role of the endocannabinoid and endovanilloid systems in an animal model of schizophrenia-related emotional processing/cognitive deficit. Neuropharmacology 155:44–53
Perez-Riverol Y, Bai J, Bandla C et al (2022) The PRIDE database resources in 2022: a hub for mass spectrometry-based proteomics evidences. Nucleic Acids Res 50:D543–D552
Funding
The authors thank FAPESP (São Paulo Research Foundation—grants 2017/18242–1, 2017/25588–1, 2018/25818–0, 2018/03673–0, 2019/00098-7), and CAPES (Coordination for the Improvement of Higher Education Personnel, grants 1656470 and 88887.495565/2020–00) for financial support.
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VA: Conceptualization, Methodology, Formal analysis, Investigation, Writing—Original Draft, Writing—Review & Editing and Funding acquisition. GS, GSZ, PR, and MF: Investigation. GRO: Formal analysis and Visualization. BJS: Writing—Review & Editing. ACC, AWZ, JEH, and JAC: Resources and Writing—Review & Editing. DMS: Conceptualization, Resources, Writing—Review & Editing, Supervision, Funding acquisition. All authors contributed to and approved the final version of the manuscript.
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JAC is a member of the International Advisory Board of the Australian Centre for Cannabinoid Clinical and Research Excellence (ACRE) – National Health and Medical Research Council (NHMRC). JAC and JEH have received travel support to attend scientific meetings and personal consultation fees from BSPG-Pharm. JAC, JEH, and AWZ are co-inventors of the patent “Fluorinated CBD compounds, compositions and uses thereof. Pub. No.: WO/2014/108899. International Application No.: PCT/IL2014/050023,” Def. US number Reg. 62193296; July 29, 2015; INPI on August 19, 2015 (BR1120150164927; Mechoulam R, Zuardi AW, Kapczinski F, Hallak JEC, Guimarães FS, Crippa JAS, Breuer A). Universidade de São Paulo (USP) has licensed this patent to Phytecs Pharm (USP Resolution No. 15.1.130002.1.1) and has an agreement with Prati-Donaduzzi to “develop a pharmaceutical product containing synthetic CBD and prove its safety and therapeutic efficacy in the treatment of epilepsy, schizophrenia, Parkinson’s disease, and anxiety disorders.” JAC, JEH, AWZ are co-inventors of the patent “Cannabinoid-containing oral pharmaceutical composition, a method for preparing and using the same,” INPI on September 16, 2016 (BR 112018005423–2).
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de Almeida, V., Seabra, G., Reis-de-Oliveira, G. et al. Cannabinoids modulate proliferation, differentiation, and migration signaling pathways in oligodendrocytes. Eur Arch Psychiatry Clin Neurosci 272, 1311–1323 (2022). https://doi.org/10.1007/s00406-022-01425-5
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DOI: https://doi.org/10.1007/s00406-022-01425-5