A phenolic small molecule inhibitor of RNase L prevents cell death from ADAR1 deficiency.
A phenolic small molecule inhibitor of RNase L prevents cell death from ADAR1 deficiency.
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RNase L 的酚类小分子抑制剂可防止 ADAR1 缺陷引起的细胞死亡
DOI：
10.1073/pnas.2006883117
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发表时间：
2020-10-06
影响因子：
11.1
通讯作者：
Sicheri F
中科院分区：
文献类型：
--
作者：
Daou S;Talukdar M;Tang J;Dong B;Banerjee S;Li Y;Duffy NM;Ogunjimi AA;Gaughan C;Jha BK;Gish G;Tavernier N;Mao D;Weiss SR;Huang H;Silverman RH;Sicheri F
OAS-RNase L系统是由病毒感染激活的先天免疫途径。病毒dsRNA刺激OAS酶产生短的2′，5 ′-寡腺苷酸（2-5A），其激活RNase L，导致单链（ss）RNA的切割。我们发现了一种RNA酶L的小分子抑制剂，可以挽救dsRNA编辑酶ADAR 1缺陷细胞的毒性表型。ADAR 1使dsRNA不稳定以阻止OAS活性。ADAR 1突变导致Aicardi-Goutières综合征（AGS）的一部分病例，AGS是一种严重的神经发育和炎症性遗传疾病，儿童没有有效的药物治疗。我们认为，RNase L抑制剂可能对RNase L被激活的AGS病例和其他RNase L过度激活有害的适应症具有效用。寡腺苷酸合成酶（OAS）-RNase L系统是由病毒感染激活的IFN诱导的抗病毒途径。病毒双链（ds）RNA激活合成第二信使2-5A的OAS同种型，所述第二信使2-5A结合并激活假激酶-核糖核酸内切酶RNase L。在细胞中，OAS的激活被ADAR 1抑制，ADAR 1是一种使dsRNA不稳定的腺苷脱氨酶。ADAR 1突变是Aicardi-Goutières综合征（AGS）的原因之一，AGS是一种儿童干扰素病。人类细胞中的ADAR 1缺陷可导致RNase L激活和随后的细胞死亡。为了评估RNase L作为AGS可能的治疗靶点，我们试图鉴定RNase L的小分子抑制剂。在500个蛋白激酶抑制剂化合物库中筛选RNA酶L活性的体外调节剂。我们确定鞣花酸（EA）作为一个命中与10倍高的选择性对RNase L相比，其最接近的paranesthesia，IRE 1。SAR分析鉴定了戊酸二内酯（瓦尔）作为RNA酶L的上级抑制剂，其选择性是IRE 1的100倍。作用机制分析表明，EA和瓦尔不结合RNase L的假激酶结构域，尽管作为蛋白激酶CK 2的ATP竞争性抑制剂。瓦尔在细胞中是无毒的和功能性的，尽管效力降低1,000倍，如通过响应于用dsRNA激活剂处理的RNA切割活性或通过拯救由ADAR 1缺陷诱导的自身dsRNA导致的细胞致死性所测量的。这些研究为理解使用小分子抑制剂调节RNase L功能的新模式和治疗潜力的途径奠定了基础。
The OAS–RNase L system is an innate immunity pathway activated by viral infection. Viral dsRNA stimulates OAS enzymes to produce short 2′,5′-oligoadenylates (2-5A) that activate RNase L, resulting in cleavage of single-stranded (ss) RNA. We discovered a small-molecule inhibitor of RNase L that rescues the toxic phenotype of cells deficient in the dsRNA-editing enzyme ADAR1. ADAR1 destabilizes dsRNA to prevent OAS activity. ADAR1 mutations are responsible for a subset of cases of Aicardi-Goutières syndrome (AGS), a severe neurodevelopmental and inflammatory genetic disease of children with no effective medical therapy. We posit that an RNase L inhibitor may have utility against cases of AGS in which RNase L is activated and other indications where overactivation of RNase L is harmful. The oligoadenylate synthetase (OAS)–RNase L system is an IFN-inducible antiviral pathway activated by viral infection. Viral double-stranded (ds) RNA activates OAS isoforms that synthesize the second messenger 2-5A, which binds and activates the pseudokinase-endoribonuclease RNase L. In cells, OAS activation is tamped down by ADAR1, an adenosine deaminase that destabilizes dsRNA. Mutation of ADAR1 is one cause of Aicardi-Goutières syndrome (AGS), an interferonopathy in children. ADAR1 deficiency in human cells can lead to RNase L activation and subsequent cell death. To evaluate RNase L as a possible therapeutic target for AGS, we sought to identify small-molecule inhibitors of RNase L. A 500-compound library of protein kinase inhibitors was screened for modulators of RNase L activity in vitro. We identified ellagic acid (EA) as a hit with 10-fold higher selectivity against RNase L compared with its nearest paralog, IRE1. SAR analysis identified valoneic acid dilactone (VAL) as a superior inhibitor of RNase L, with 100-fold selectivity over IRE1. Mechanism-of-action analysis indicated that EA and VAL do not bind to the pseudokinase domain of RNase L despite acting as ATP competitive inhibitors of the protein kinase CK2. VAL is nontoxic and functional in cells, although with a 1,000-fold decrease in potency, as measured by RNA cleavage activity in response to treatment with dsRNA activator or by rescue of cell lethality resulting from self dsRNA induced by ADAR1 deficiency. These studies lay the foundation for understanding novel modes of regulating RNase L function using small-molecule inhibitors and avenues of therapeutic potential.
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影响因子：
5.2
作者：
De, Ronita;Sarkar, Avijit;Mukhopadhyay, Asish K.
通讯作者：
Mukhopadhyay, Asish K.
DOI：
10.1073/pnas.1519657113
发表时间：
2016-02-23
影响因子：
11.1
作者：
Li, Yize;Banerjee, Shuvojit;Weiss, Susan R.
通讯作者：
Weiss, Susan R.
DOI：
10.1073/pnas.1815071116
发表时间：
2019-03-12
影响因子：
11.1
作者：
Banerjee, Shuvojit;Gusho, Elona;Silverman, Robert H.
通讯作者：
Silverman, Robert H.
影响因子：
6.1
作者：
Li, Li;Tsao, Rong;Young, J. Christopher
通讯作者：
Young, J. Christopher
影响因子：
9.7
作者：
Krueger, Stefan;Engel, Christoph;Schackert, Hans K.
通讯作者：
Schackert, Hans K.