Selective Functional Inhibition of JAK-3 Is Sufficient for Efficacy in Collagen-Induced Arthritis in Mice
Selective Functional Inhibition of JAK-3 Is Sufficient for Efficacy in Collagen-Induced Arthritis in Mice
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DOI：
10.1002/art.27536
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发表时间：
2010-08-01
影响因子：
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通讯作者：
Seidl, Katherine J.
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文献类型：
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作者：
Lin, Tsung H.;Hegen, Martin;Seidl, Katherine J.
目标。所有的γ链细胞因子通过JAK-3和JAK-1串联作用来传递信号。我们进行了这项研究，以确定JAK-3选择性抑制剂WYE-151650是否足以破坏细胞因子信号并改善自身免疫性疾病病理，而不抑制由JAK-1、JAK-2和tyk -2介导的其他途径。JAK-3激酶选择性化合物通过激酶测定和JAK-3依赖性（白介素-2 [IL-2]）和非依赖性（IL-6，粒细胞-巨噬细胞集落刺激因子[GM-CSF]）细胞测定增殖或STAT磷酸化来表征。在体内，脱靶信号通过IL-22和促红细胞生成素（EPO）介导的模型来测量，而靶信号通过il -2介导的信号来测量。采用小鼠迟发性超敏反应（DTH）和胶原诱导关节炎（CIA）模型研究JAK-3抑制剂的疗效。在体外，WYE-151650能有效抑制il -2诱导的STAT-5磷酸化和细胞增殖，而对jak -3独立的IL-6-或gm - csf诱导的STAT磷酸化的抑制活性降低10-29倍。在体外，WYE-151650在全血中抑制il -2诱导的STAT磷酸化，但不抑制il -6诱导的STAT磷酸化。在体内，WYE-151650抑制了jak -3介导的il -2诱导的干扰素γ的产生，减少了小鼠的自然杀伤细胞数量，而不影响il -22诱导的血清淀粉样蛋白A的产生或epo诱导的网状细胞增多症。WYE-151650对小鼠DTH和CIA模型均有效。体外、离体和体内实验表明，尽管有JAK-3选择性，但WYE-151650对小鼠CIA有效。这些数据质疑是否需要广泛抑制JAK-1-、JAK-2-或tyk -2依赖性细胞因子途径来获得疗效。
Objective. All gamma-chain cytokines signal through JAK-3 and JAK-1 acting in tandem. We undertook this study to determine whether the JAK-3 selective inhibitor WYE-151650 would be sufficient to disrupt cytokine signaling and to ameliorate autoimmune disease pathology without inhibiting other pathways mediated by JAK-1, JAK-2, and Tyk-2.Methods. JAK-3 kinase selective compounds were characterized by kinase assay and JAK-3-dependent (interleukin-2 [IL-2]) and -independent (IL-6, granulocyte-macrophage colony-stimulating factor [GM-CSF]) cell-based assays measuring proliferation or STAT phosphorylation. In vivo, off-target signaling was measured by IL-22- and erythropoietin (EPO)-mediated models, while on-target signaling was measured by IL-2-mediated signaling. Efficacy of JAK-3 inhibitors was determined using delayed-type hypersensitivity (DTH) and collagen-induced arthritis (CIA) models in mice.Results. In vitro, WYE-151650 potently suppressed IL-2-induced STAT-5 phosphorylation and cell proliferation, while exhibiting 10-29-fold less activity against JAK-3-independent IL-6- or GM-CSF-induced STAT phosphorylation. Ex vivo, WYE-151650 suppressed IL-2-induced STAT phosphorylation, but not IL-6-induced STAT phosphorylation, as measured in whole blood. In vivo, WYE-151650 inhibited JAK-3-mediated IL-2-induced interferon-gamma production and decreased the natural killer cell population in mice, while not affecting IL-22-induced serum amyloid A production or EPO-induced reticulocytosis. WYE-151650 was efficacious in mouse DTH and CIA models.Conclusion. In vitro, ex vivo, and in vivo assays demonstrate that WYE-151650 is efficacious in mouse CIA despite JAK-3 selectivity. These data question the need to broadly inhibit JAK-1-, JAK-2-, or Tyk-2-dependent cytokine pathways for efficacy.