Blockade of TANK-Binding Kinase 1/IKKε Inhibits Mutant Stimulator of Interferon Genes (STING)-Mediated Inflammatory Responses in Human Peripheral Blood Mononuclear Cells
Blockade of TANK-Binding Kinase 1/IKKε Inhibits Mutant Stimulator of Interferon Genes (STING)-Mediated Inflammatory Responses in Human Peripheral Blood Mononuclear Cells
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DOI：
10.1002/art.40122
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发表时间：
2017-07-01
影响因子：
13.3
通讯作者：
Rodero, Mathieu P.
中科院分区：
文献类型：
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作者：
Fremond, Marie-Louise;Uggenti, Carolina;Rodero, Mathieu P.
目标。编码干扰素刺激因子（IFN）基因（STING）蛋白的TMEM173的功能获得突变是一种新型I型干扰素病的基础，这种病对常规免疫抑制疗法的反应最低，并且与儿童发病率和死亡率的高频率相关。STING的功能获得导致IFN的组成性过度分泌。本研究旨在探讨TANK-binding kinase 1 (TBK-1)/IKKE抑制剂（BX795）对sting突变患者外周血单个核细胞（PBMCs） IFN分泌和信号转导的影响。用BX795治疗4例sting相关疾病患者的PBMCs。BX795对IFN通路的影响通过Western blotting和IFN β报告基因法，以及细胞裂解物中IFN α的定量，STAT-1磷酸化染色和IFN刺激基因（ISG）信使RNA （mRNA）表达的测量来评估。用BX795处理PBMCs可抑制环GMP-AMP或基因激活STING诱导的HEK 293T细胞中IFN调节因子3的磷酸化和IFNb启动子活性。体外暴露于BX795可抑制sting相关疾病患者pbmc中IFNa的产生，但不影响细胞存活。此外，BX795降低STAT-1磷酸化和ISG mRNA表达，不依赖于IFNa阻断。这些发现证明了BX795在STING功能获得突变患者细胞中减少I型IFN产生和IFN信号传导的作用。因此，联合抑制TBK-1和IKKE具有治疗携带STING突变的患者的潜力，也可能与其他I型干扰素病变有关。
Objective. Gain-of-function mutations in TMEM173, encoding the stimulator of interferon (IFN) genes (STING) protein, underlie a novel type I interferonopathy that is minimally responsive to conventional immunosuppressive therapies and associated with high frequency of childhood morbidity and mortality. STING gain-of-function causes constitutive oversecretion of IFN. This study was undertaken to determine the effects of a TANK-binding kinase 1 (TBK-1)/IKKE inhibitor (BX795) on secretion and signaling of IFN in primary peripheral blood mononuclear cells (PBMCs) from patients with mutations in STING.Methods. PBMCs from 4 patients with STING-associated disease were treated with BX795. The effect of BX795 on IFN pathways was assessed by Western blotting and an IFN beta reporter assay, as well as by quantification of IFN alpha in cell lysates, staining for STAT-1 phosphorylation, and measurement of IFN-stimulated gene (ISG) messenger RNA (mRNA) expression.Results. Treatment of PBMCs with BX795 inhibited the phosphorylation of IFN regulatory factor 3 and IFNb promoter activity induced in HEK 293T cells by cyclic GMP-AMP or by genetic activation of STING. In vitro exposure to BX795 inhibited IFNa production in PBMCs of patients with STING-associated disease without affecting cell survival. In addition, BX795 decreased STAT-1 phosphorylation and ISG mRNA expression independent of IFNa blockade.Conclusion. These findings demonstrate the effect of BX795 on reducing type I IFN production and IFN signaling in cells from patients with gain-of-function mutations in STING. A combined inhibition of TBK-1 and IKKE therefore holds potential for the treatment of patients carrying STING mutations, and may also be relevant in other type I interferonopathies.