Blockade of USP14 potentiates type I interferon signaling and radiation-induced antitumor immunity via preventing IRF3 deubiquitination
Blockade of USP14 potentiates type I interferon signaling and radiation-induced antitumor immunity via preventing IRF3 deubiquitination
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DOI：
10.1007/s13402-022-00724-2
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发表时间：
2022-10
影响因子：
6.6
通讯作者：
Weijian Wu;Huiying Xu;C. Liao;Youqiao Wang;Ruirui Wu;Jiaxin Wu;Wenlv Zheng;Yun-Yi Li;
中科院分区：
文献类型：
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作者：
Weijian Wu;Huiying Xu;C. Liao;Youqiao Wang;Ruirui Wu;Jiaxin Wu;Wenlv Zheng;Yun-Yi Li;
目的放疗诱导的适应性免疫反应在很大程度上依赖于sting依赖性I型干扰素（ifn）的产生。然而，辐照肿瘤细胞往往不能诱导树突状细胞（dc）产生I型ifn。因此，我们的目标是揭示sting介导的放射后先天免疫感知的局限性，并确定能够挽救I型IFNs诱导失败的有效试剂，以促进放射治疗。方法采用靶向细胞表型筛选方法，寻找能促进I型ifn产生的活性分子。体外测试USP14敲除或抑制对IFN产生和STING信号激活的影响。采用western blot和体外免疫共沉淀法研究USP14的作用机制。此外，在体内和体外模型中进行PT33和辐射联合治疗，以评估I型IFNs对辐射的反应。结果spt33被鉴定为STING激动剂诱导的I型ifn产生的增强剂，从而在体外产生高水平和持久的STING激活谱。从机制上讲，USP14的抑制或缺失会损害K63-linked IRF3的去泛素化。此外，PT33阻断USP14可以增强体外照射肿瘤细胞的DC感知，并与辐射协同促进体内全身抗肿瘤免疫。结论USP14是主要的IFN产生抑制因子之一，并通过去除IRF3的k63连锁泛素化而损害IRF3的激活。因此，阻断USP14可获得STING信号激活和辐射诱导的适应性免疫应答。
PurposeThe adaptive immune responses induced by radiotherapy has been demonstrated to largely rely on STING-dependent type I interferons (IFNs) production. However, irradiated tumor cells often fail to induce dendritic cells (DCs) to produce type I IFNs. Hence, we aim to uncover the limitation of STING-mediated innate immune sensing following radiation, and identify efficient reagents capable to rescue the failure of type I IFNs induction for facilitating radiotherapy.MethodsA targeted cell-based phenotypic screening was performed to search for active molecules that could elevate the production of type I IFNs. USP14 knockout or inhibition was assayed for IFN production and the activation of STING signaling in vitro. The mechanisms of USP14 were investigated by western blot and co-immunoprecipitation in vitro. Additionally, combinational treatments with PT33 and radiation in vivo and in vitro models were performed to evaluate type I IFNs responses to radiation.ResultsPT33 was identified as an enhancer of STING agonist elicited type I IFNs production to generate an elevated and durable STING activation profile in vitro. Mechanistically, USP14 inhibition or deletion impairs the deubiquitylation of K63-linked IRF3. Furthermore, blockade of USP14 with PT33 enhances DC sensing of irradiated-tumor cells in vitro, and synergizes with radiation to promote systemic antitumor immunity in vivo.ConclusionOur findings reveal that USP14 is one of the major IFN production suppressors and impairs the activation of IRF3 by removing the K63-linked ubiquitination of IRF3. Therefore, blockage of USP14 results in the gain of STING signaling activation and radiation-induced adaptive immune responses.