title: The peripheral cannabinoid receptor Cb2, a novel oncoprotein, induces a reversible block in neutrophilic differentiation.
abstract: We previously identified a novel common virus integration site, Evi11, by means of retroviral insertional mutagenesis. We demonstrated that the gene encoding the peripheral cannabinoid receptor (Cb2) is the potential target, suggesting that Cb2 is a proto-oncogene. To elucidate a role for this G protein-coupled receptor (GPCR) in leukemic transformation we generated a Cb2-EGFP cDNA construct that was introduced into 32D/G-CSF-R cells. These cells require interleukin 3 (IL-3) to proliferate in vitro, whereas in the presence of granulocyte-colony-stimulating factor (G-CSF) they differentiate toward mature neutrophils. We demonstrate that 32D/G-CSF-R/Cb2-EGFP cells migrate in a transwell assay in reponse to the Cb2 ligand 2-arachidonoylglycerol (2-AG), indicating that the fusion protein was functional. When cultured in the presence of G-CSF neutrophilic differentiation of Cb2-EGFP-expressing 32D/G-CSF-R cells was completely blocked. Moreover, a Cb2-specific antagonist fully recovered the G-CSF-induced neutrophilic differentiation of 32D/G-CSF-R/Cb2-EGFP cells. To investigate which signal transduction pathway(s) may be involved in the block of neutrophilic maturation, differentiation experiments were carried out using specific inhibitors of signaling routes. Interestingly, full rescue of G-CSF-induced neutrophilic differentiation was observed when cells were cultured with the mitogen-induced extracellular kinase (MEK) inhibitors, PD98059 or U0126, and partial recovery was detected with the phosphoinositide 3-kinase (PI3-K) inhibitor LY-294002. These studies demonstrate that the Cb2 receptor is an oncoprotein that blocks neutrophilic differentiation when overexpressed in myeloid precursor cells. Cb2 appears to mediate its activity through MEK/extracellular signal-related kinase (ERK) and PI3-K pathways.
authors.affiliation: The Institute for Hematology, Erasmus Medical Center, The Netherlands.
authors.forename: Meritxell Alberich
authors.initials: MA
authors.lastname: Jordà
authors.forename: Bob
authors.initials: B
authors.lastname: Lowenberg
authors.forename: Ruud
authors.initials: R
authors.lastname: Delwel
mesh_terms: D001095:Arachidonic Acids; D002070:Butadienes / Q000494:pharmacology; D002454:Cell Differentiation / Q000502:physiology*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002634:Chemotaxis, Leukocyte; D002867:Chromones / Q000494:pharmacology; D063388:Endocannabinoids; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D005989:Glycerides; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors; D009025:Morpholines / Q000494:pharmacology; D009504:Neutrophils / Q000166:cytology*; D009570:Nitriles / Q000494:pharmacology; D015513:Oncogene Proteins / Q000502:physiology*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D000090063:Proto-Oncogene Mas; D043882:Receptors, Cannabinoid; D011955:Receptors, Drug / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins; D015398:Signal Transduction
chemical_list: D001095:Arachidonic Acids; D002070:Butadienes; D002867:Chromones; D063388:Endocannabinoids; D004791:Enzyme Inhibitors; D005419:Flavonoids; D005989:Glycerides; D007377:Interleukin-3; C000715470:MAS1 protein, human; D009025:Morpholines; D009570:Nitriles; D015513:Oncogene Proteins; D000081082:Phosphoinositide-3 Kinase Inhibitors; D000090063:Proto-Oncogene Mas; D043882:Receptors, Cannabinoid; D011955:Receptors, Drug; D011993:Recombinant Fusion Proteins; C113580:U 0126; D016179:Granulocyte Colony-Stimulating Factor; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; C094503:glyceryl 2-arachidonate; D020928:Mitogen-Activated Protein Kinases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one