title: Activation of T cell death-associated gene 8 regulates the cytokine production of T cells and macrophages in vitro.
abstract: An orphan G-protein-coupled receptor, T cell death-associated gene 8 (TDAG8) which has been reported to be a proton sensor, inhibits the production of pro-inflammatory cytokines induced by extracellular acidification. Recently, we have found that TDAG8 knockout mice showed significant exacerbation in various immune-mediated inflammation disease models. To elucidate the role of TDAG8, we screened an in-house library to find compounds which have a profile as a TDAG8 agonist using a cyclic adenosine 5'-monophosphate assay. Among the screening hits, we focused on (3-[(2,4-dichlorobenzyl)thio]-1,6-dimethyl-5,6-dihydro-1H-pyridazino[4,5-e][1,3,4]thiadiazin-5-one) (named BTB09089). BTB09089 did not act on other proton sensing G-protein-coupled receptors such as G-protein-coupled receptor 4 (GPR4) nor ovarian cancer G-protein-coupled receptor 1 (OGR1). Moreover, BTB09089 increased cAMP level in the splenocytes from wild-type littermates but not from TDAG8-deficient mice. Thus, BTB09089 was found to be a TDAG8 specific agonist. We then investigated the effects of BTB09089 on T cells and macrophages in vitro. In splenocytes, BTB09089 suppressed the production of IL-2 stimulated with anti-CD3 and anti-CD28 antibodies. In peritoneal exuded macrophages induced by thioglycollate, BTB09089 suppressed the production of TNF-α and IL-6 while it increased that of IL-10 when stimulated with lipopolysaccharide. These effects were observed in cells from wild type mice, but not those from TDAG8 knockout mice. These results indicate that activation of TDAG8 attenuates immune-mediated inflammation by regulating the cytokine production of T cells and macrophages.
authors.affiliation: Biological Research Laboratories III, Daiichi Sankyo Co., Ltd., 1-16-13 Kitakasai, Edogawa-ku, Tokyo 134-8630, Japan.
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Onozawa
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Fujita
authors.forename: Harumi
authors.initials: H
authors.lastname: Kuwabara
authors.forename: Miyuki
authors.initials: M
authors.lastname: Nagasaki
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Komai
authors.forename: Tomiichiro
authors.initials: T
authors.lastname: Oda
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D016207:Cytokines / Q000378:metabolism*; D055808:Drug Discovery; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D011724:Pyridazines / Q000494:pharmacology; D043562:Receptors, G-Protein-Coupled / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000819:agonists / Q000378:metabolism; D054852:Small Molecule Libraries / Q000494:pharmacology; D013154:Spleen / Q000166:cytology / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D013829:Thiadiazines / Q000494:pharmacology
chemical_list: C574797:3-((2,4-dichlorobenzyl)thio)-1,6-dimethyl-5,6-dihydro-1H-pyridazino(4,5-e)(1,3,4)thiadiazin-5-one; D000894:Anti-Inflammatory Agents, Non-Steroidal; D016207:Cytokines; C513334:GPR65 protein, human; C513337:GPR65 protein, mouse; D011724:Pyridazines; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins; D054852:Small Molecule Libraries; D013829:Thiadiazines