cAbl Kinase Regulates Inflammasome Activation and Pyroptosis via ASC Phosphorylation.
cAbl Kinase Regulates Inflammasome Activation and Pyroptosis via ASC Phosphorylation.
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DOI：
10.4049/jimmunol.2000969
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发表时间：
2021-03-15
期刊：
影响因子：
--
通讯作者：
Wewers MD
中科院分区：
文献类型：
--
作者：
Gavrilin MA;Prather ER;Vompe AD;McAndrew CC;Wewers MD
炎性体激活部分地通过炎性体蛋白的翻译后修饰来调节。酪氨酸磷酸化是一种可能的修饰。先前已经显示蛋白酪氨酸激酶（PTK）抑制剂AG 126极大地抑制炎性小体活化，我们试图揭示靶激酶。为此，我们筛选了商业酪氨酸激酶文库以抑制炎性小体依赖性IL-18/IL-1β释放和细胞凋亡。THP-1细胞（人单核细胞系）与PTK抑制剂（0.1、1和10 μM）一起孵育，然后用LPS刺激，然后用ATP刺激。PTK抑制剂DCC-2036（Rebastinib）和GZD 824（对Bcr-Abl激酶具有特异性）在所有使用浓度下均显示出IL-18和LDH释放的最严重减少。然后通过CRISPR/Cas9编辑在THP-1细胞中删除所建议的激酶靶点cAbl激酶，然后测试其在炎性小体功能中的作用和使炎性小体衔接子ASC磷酸化的潜力。cABL KO不仅显著抑制炎性小体功能，而且减少LPS/ATP刺激后磷酸化ASC的释放。cAbl激酶的一个预测靶标是ASC中的酪氨酸146。与野生型ASC互补相比，用突变的Y146 A ASC互补ASC KO THP-1细胞显著消除了炎性小体活化和ASC寡聚化。因此，这些发现支持cAbl激酶作为炎性小体活性和焦亡的正调节剂，可能通过ASC的磷酸化。
Inflammasome activation is regulated in part by the post-translational modification of inflammasome proteins. Tyrosine phosphorylation is one possible modification. Having previously shown that the protein tyrosine kinase (PTK) inhibitor AG126 greatly inhibits inflammasome activation, we sought to uncover the target kinase. To do this we screened a commercial tyrosine kinase library for inhibition of inflammasome-dependent IL-18/IL-1β release and pyroptosis. THP-1 cells (human monocyte cell line) were incubated with PTK inhibitors (0.1, 1 and 10 μM) before stimulation with LPS followed by ATP. The PTK inhibitors DCC-2036 (Rebastinib) and GZD824, specific for Bcr-Abl kinase, showed the most severe reduction of IL-18 and LDH release at all concentrations used. The suggested kinase target, cAbl kinase, was then deleted in THP-1 cells by CRISPR/Cas9 editing and then tested for its role in inflammasome function and potential to phosphorylate the inflammasome adaptor ASC. The cABL KO not only significantly inhibited inflammasome function but also decreased release of phosphorylated ASC after LPS/ATP stimulation. One predicted target of cAbl kinase is tyrosine 146 in ASC. Complementation of ASC KO THP-1 cells with mutated Y146A ASC significantly abrogated inflammasome activation and ASC oligomerization as compared to wild type ASC complementation. Thus, these findings support cAbl kinase as a positive regulator of inflammasome activity and pyroptosis, likely via phosphorylation of ASC.
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影响因子：
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发表时间：
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影响因子：
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