title: Pharmacological inhibition of glycogen synthase kinase 3 regulates T cell development in vitro.
abstract: The development of functional T cells requires receptor-mediated transition through multiple checkpoints in the thymus. Double negative 3 (DN3) thymocytes are selected for the presence of a rearranged TCR beta chain in a process termed β-selection which requires signalling via the pre-TCR, Notch1 and CXCL12. Signal integration by these receptors converges on core pathways including the Phosphatidylinositol-3-kinase (PI3K) pathway. Glycogen Synthase Kinase 3 (GSK3) is generally thought to be negatively regulated by the PI3K pathway but its role in β-selection has not been characterised. Here we show that developmental progression of DN3 thymocytes is promoted following inhibition of GSK3 by the synthetic compound CHIR99021. CHIR99021 allows differentiation in the absence of pre-TCR-, Notch1- or CXCL12-mediated signalling. It antagonizes IL-7-mediated inhibition of DP thymocyte differentiation and increases IL-7-promoted cell recovery. These data indicate a potentially important role for inactivation of GSK3 during β-selection. They might help to establish an in vitro stromal cell-free culture system of thymocyte development and offer a new platform for screening regulators of proliferation, differentiation and apoptosis.
authors.affiliation: Laboratory of Lymphocyte Signalling and Development, The Babraham Institute, Babraham Research Campus, Cambridge, United Kingdom.
authors.forename: Jan-Hendrik
authors.initials: JH
authors.lastname: Schroeder
authors.forename: Lewis S
authors.initials: LS
authors.lastname: Bell
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Janas
authors.forename: Martin
authors.initials: M
authors.lastname: Turner
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors*; D015851:Interleukin-7 / Q000378:metabolism; D053699:Interleukin-7 Receptor alpha Subunit / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D011725:Pyridines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D051881:Receptor, Notch1 / Q000378:metabolism; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000276:immunology*
chemical_list: C473711:Chir 99021; D004791:Enzyme Inhibitors; D015851:Interleukin-7; D053699:Interleukin-7 Receptor alpha Subunit; C497762:Notch1 protein, mouse; D011725:Pyridines; D011743:Pyrimidines; D051881:Receptor, Notch1; D016693:Receptors, Antigen, T-Cell, alpha-beta; D038362:Glycogen Synthase Kinase 3