Autoinflammatory mutation in NLRC4 reveals a leucine-rich repeat (LRR)-LRR oligomerization interface.
Autoinflammatory mutation in NLRC4 reveals a leucine-rich repeat (LRR)-LRR oligomerization interface.
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NLRC4 的自身炎症突变揭示了富含亮氨酸重复序列 (LRR)-LRR 寡聚化界面
DOI：
10.1016/j.jaci.2018.04.033
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发表时间：
2018-12
期刊：
影响因子：
--
通讯作者：
Masters SL
中科院分区：
文献类型：
--
作者：
Moghaddas F;Zeng P;Zhang Y;Schützle H;Brenner S;Hofmann SR;Berner R;Zhao Y;Lu B;Chen X;Zhang L;Cheng S;Winkler S;Lehmberg K;Canna SW;Czabotar PE;Wicks IP;De Nardo D;Hedrich CM;Zeng H;Masters SL
单基因自身炎性病症的特征在于先天性免疫系统的失调，例如通过炎性小体形成蛋白（如含NOD样受体家族CARD的4蛋白（NLRC 4））中的功能获得性突变。在这里，我们研究了NLRC 4（c.G1965C，p.W655C）富含亮氨酸重复序列（LRR）结构域中的一种新突变导致自身炎症性疾病的机制。方法：我们研究了2例不相关的早发性巨噬细胞活化综合征患者，他们在NLRC 4中携带相同的从头突变。通过使用流式细胞术分析包含半胱天冬酶募集结构域（ASC）斑点的骨化相关斑点样蛋白定量体外炎性小体复合物形成。使用重复的规则间隔短回文重复序列（CRISPR）/Cas9技术和慢病毒转导来产生具有野生型或突变型NLRC 4 cDNA的THP-1细胞。分别采用流式细胞术和ELISA定量细胞死亡和IL-1β/IL-18释放。p.W655C NLRC 4突变导致ASC斑点形成、caspase-1依赖性细胞死亡和IL-1β/IL-18产生增加。ASC促进p.W655C NLRC 4介导的细胞因子释放，但不促进细胞死亡。p.W655的突变通过与寡聚体的相对LRR结构域上的2个界面接合来激活NLRC 4炎性体复合物。一组关键的残基（p.D1010、p.D1011、p.L1012和p.I1015）在由突变型NLRC 4或NLRC 4炎性体复合物的3型分泌系统效应物（PrgI）刺激触发时参与LRR-LRR寡聚化。这是第一个报告的突变在LRR域的NLRC 4引起自身炎症性疾病。c.G1965C/p.W655C NLRC 4在体外增加炎性小体活化。从各种NLRC 4突变产生的数据提供了证据，表明LRR-LRR界面在NLRC 4炎性体复合物的寡聚化中具有重要且先前未被认识的作用。
Monogenic autoinflammatory disorders are characterized by dysregulation of the innate immune system, for example by gain-of-function mutations in inflammasome-forming proteins, such as NOD-like receptor family CARD-containing 4 protein (NLRC4). Here we investigate the mechanism by which a novel mutation in the leucine-rich repeat (LRR) domain of NLRC4 (c.G1965C, p.W655C) contributes to autoinflammatory disease. Methods: We studied 2 unrelated patients with early-onset macrophage activation syndrome harboring the same de novo mutation in NLRC4. In vitro inflammasome complex formation was quantified by using flow cytometric analysis of apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) specks. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 techniques and lentiviral transduction were used to generate THP-1 cells with either wild-type or mutant NLRC4 cDNA. Cell death and release of IL-1β/IL-18 were quantified by using flow cytometry and ELISA, respectively. The p.W655C NLRC4 mutation caused increased ASC speck formation, caspase-1–dependent cell death, and IL-1β/IL-18 production. ASC contributed to p.W655C NLRC4–mediated cytokine release but not cell death. Mutation of p.W655 activated the NLRC4 inflammasome complex by engaging with 2 interfaces on the opposing LRR domain of the oligomer. One key set of residues (p.D1010, p.D1011, p.L1012, and p.I1015) participated in LRR-LRR oligomerization when triggered by mutant NLRC4 or type 3 secretion system effector (PrgI) stimulation of the NLRC4 inflammasome complex. This is the first report of a mutation in the LRR domain of NLRC4 causing autoinflammatory disease. c.G1965C/p.W655C NLRC4 increased inflammasome activation in vitro. Data generated from various NLRC4 mutations provides evidence that the LRR-LRR interface has an important and previously unrecognized role in oligomerization of the NLRC4 inflammasome complex.
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影响因子：
1.1
作者：
Hardjo Lugito NP;Cucunawangsih;Kurniawan A
通讯作者：
Kurniawan A
影响因子：
14.8
作者：
Kumar, Prateek;Henikoff, Steven;Ng, Pauline C.
通讯作者：
Ng, Pauline C.
DOI：
10.1073/pnas.0913087107
发表时间：
2010-02-16
影响因子：
11.1
作者：
Miao, Edward A.;Mao, Dat P.;Aderem, Alan
通讯作者：
Aderem, Alan
影响因子：
1.9
作者：
Liang, Jiancong;Alfano, Danielle N.;Picarsic, Jennifer
通讯作者：
Picarsic, Jennifer
影响因子：
2.3
作者：
Mutlu, Mehmet;Bayramoglu, Gulcin;Aslan, Yakup
通讯作者：
Aslan, Yakup