ARPC1B promotes mesenchymal phenotype maintenance and radiotherapy resistance by blocking TRIM21-mediated degradation of IFI16 and HuR in glioma stem cells.
ARPC1B promotes mesenchymal phenotype maintenance and radiotherapy resistance by blocking TRIM21-mediated degradation of IFI16 and HuR in glioma stem cells.
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ARPC1B 通过阻断 TRIM21 介导的胶质瘤干细胞中 IFI16 和 HuR 的降解来促进间充质表型维持和放射治疗抵抗
DOI：
10.1186/s13046-022-02526-8
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发表时间：
2022-11-16
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影响因子：
--
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瘤内异质性是胶质母细胞瘤（GBM）治疗的主要挑战。神经胶质瘤干细胞（GSC）的存在及其在不同分子表型之间的转换导致了异质性的复杂性，最终导致对放射治疗的优先抵抗。 ARP2/3（肌动蛋白相关蛋白2/3）复合物（ARPs）与癌症迁移、侵袭和分化相关，但ARPs在GSC表型和放疗抵抗中的影响仍不清楚。我们筛选了 TCGA-GBM 和 CGGA-GBM 数据库中的 ARP 表达。应用肿瘤球形成测定和有限稀释测定来评估 ARPC1B 在肿瘤发生中的影响。采用细胞凋亡、彗星、γ-H2AX 免疫荧光 (IF) 和细胞周期分布测定来评估 ARPC1B 对放疗抵抗的影响。使用免疫沉淀 (IP) 和质谱分析来检测 ARPC1B 相互作用蛋白。进行免疫印迹测定来评估蛋白质泛素化，并设计缺失突变体构建体来确定蛋白质相互作用的结合位点。执行 Spearman 相关算法来筛选通过 ARPC1B 的表达指示细胞敏感性的药物。使用颅内异种移植 GSC 小鼠模型来研究 ARPC1B 在体内的作用。我们得出结论，ARPC1B 在 MES-GBM/GSC 中显着上调，并且与不良预后相关。体外和体内试验均表明，MES-GSC 中 ARPC1B 的敲低可降低致瘤性和对 IR 治疗的耐药性，而 PN-GSC 中 ARPC1B 的过表达则表现出相反的效果。从机制上讲，ARPC1B 与 IFI16 和 HuR 相互作用以维持蛋白质稳定性。详细而言，IFI16 的 Pyrin 和 HuR 的 RRM2 与 ARPC1B 的结合有关，这抵消了 TRIM21 介导的 IFI16 和 HuR 泛素化降解。此外，ARPC1B 的功能依赖于 IFI16 诱导的 NF-κB 通路激活和 HuR 诱导的 STAT3 通路激活。最后，我们根据ARPC1B的表达筛选了AZD6738，一种共济失调毛细血管扩张突变的rad3相关（ATR）抑制剂。除了反映细胞对 AZD6738 敏感性的 ARPC1B 表达之外，AZD6738 与放射疗法的组合在体外和体内均表现出有效的抗肿瘤作用。 ARPC1B 通过抑制 TRIM21 介导的 IFI16 和 HuR 降解，从而分别激活 NF-κB 和 STAT3 信号通路，促进 MES 表型维持和放疗抵抗。根据 ARPC1B 表达鉴定的 AZD6738 与放射治疗相结合，表现出优异的抗 GSC 活性。在线版本包含可在 10.1186/s13046-022-02526-8 获取的补充材料。
Intratumoral heterogeneity is the primary challenge in the treatment of glioblastoma (GBM). The presence of glioma stem cells (GSCs) and their conversion between different molecular phenotypes contribute to the complexity of heterogeneity, culminating in preferential resistance to radiotherapy. ARP2/3 (actin-related protein-2/3) complexes (ARPs) are associated with cancer migration, invasion and differentiation, while the implications of ARPs in the phenotype and resistance to radiotherapy of GSCs remain unclear. We screened the expression of ARPs in TCGA-GBM and CGGA-GBM databases. Tumor sphere formation assays and limiting dilution assays were applied to assess the implications of ARPC1B in tumorigenesis. Apoptosis, comet, γ-H2AX immunofluorescence (IF), and cell cycle distribution assays were used to evaluate the effect of ARPC1B on radiotherapy resistance. Immunoprecipitation (IP) and mass spectrometry analysis were used to detect ARPC1B-interacting proteins. Immune blot assays were performed to evaluate protein ubiquitination, and deletion mutant constructs were designed to determine the binding sites of protein interactions. The Spearman correlation algorithm was performed to screen for drugs that indicated cell sensitivity by the expression of ARPC1B. An intracranial xenograft GSC mouse model was used to investigate the role of ARPC1B in vivo. We concluded that ARPC1B was significantly upregulated in MES-GBM/GSCs and was correlated with a poor prognosis. Both in vitro and in vivo assays indicated that knockdown of ARPC1B in MES-GSCs reduced tumorigenicity and resistance to IR treatment, whereas overexpression of ARPC1B in PN-GSCs exhibited the opposite effects. Mechanistically, ARPC1B interacted with IFI16 and HuR to maintain protein stability. In detail, the Pyrin of IFI16 and RRM2 of HuR were implicated in binding to ARPC1B, which counteracted TRIM21-mediated degradation of ubiquitination to IFI16 and HuR. Additionally, the function of ARPC1B was dependent on IFI16-induced activation of NF-κB pathway and HuR-induced activation of STAT3 pathway. Finally, we screened AZD6738, an ataxia telangiectasia mutated and rad3-related (ATR) inhibitor, based on the expression of ARPC1B. In addition to ARPC1B expression reflecting cellular sensitivity to AZD6738, the combination of AZD6738 and radiotherapy exhibited potent antitumor effects both in vitro and in vivo. ARPC1B promoted MES phenotype maintenance and radiotherapy resistance by inhibiting TRIM21-mediated degradation of IFI16 and HuR, thereby activating the NF-κB and STAT3 signaling pathways, respectively. AZD6738, identified based on ARPC1B expression, exhibited excellent anti-GSC activity in combination with radiotherapy. The online version contains supplementary material available at 10.1186/s13046-022-02526-8.
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ATM 和 IFI16 对 DNA 传感适配器 STING 的非规范激活可在核 DNA 损伤后介导 NF-κB 信号转导。
影响因子：
16
作者：
Dunphy G;Flannery SM;Almine JF;Connolly DJ;Paulus C;Jønsson KL;Jakobsen MR;Nevels MM;Bowie AG;Unterholzner L
通讯作者：
Unterholzner L
影响因子：
50.3
作者：
Hara T;Chanoch-Myers R;Mathewson ND;Myskiw C;Atta L;Bussema L;Eichhorn SW;Greenwald AC;Kinker GS;Rodman C;Gonzalez Castro LN;Wakimoto H;Rozenblatt-Rosen O;Zhuang X;Fan J;Hunter T;Verma IM;Wucherpfennig KW;Regev A;Suvà ML;Tirosh I
通讯作者：
Tirosh I
影响因子：
11.2
作者：
Kazazian, Karineh;Go, Christopher;Swallow, Carol J.
通讯作者：
Swallow, Carol J.
影响因子：
7.3
作者：
Foote, Kevin M.;Nissink, J. Willem M.;Jewsbury, Philip J.
通讯作者：
Jewsbury, Philip J.
DOI：
10.1158/1078-0432.ccr-21-0251
发表时间：
2021-09-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Kim ST;Smith SA;Mortimer P;Loembé AB;Cho H;Kim KM;Smith C;Willis S;Irurzun-Arana I;Berges A;Hong JY;Park SH;Park JO;Park YS;Lim HY;Kang WK;Kozarewa I;Pierce AJ;Dean E;Lee J
通讯作者：
Lee J