TY  - JOUR
AU  - Moghaddas, F
AU  - Zeng, P
AU  - Zhang, Y
AU  - Schuetzle, H
AU  - Brenner, S
AU  - Hofmann, SR
AU  - Berner, R
AU  - Zhao, Y
AU  - Lu, B
AU  - Chen, X
AU  - Zhang, L
AU  - Cheng, S
AU  - Winkler, S
AU  - Lehmberg, K
AU  - Canna, SW
AU  - Czabotar, PE
AU  - Wicks, IP
AU  - De Nardo, D
AU  - Hedrich, CM
AU  - Zeng, H
AU  - Masters, SL
Y2  - 2020/12/17
Y1  - 2018/12/01
SN  - 0091-6749
UR  - http://hdl.handle.net/11343/255350
AB  - BACKGROUND: 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). OBJECTIVE: 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. RESULTS: 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. CONCLUSION: 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.
LA  - English
PB  - MOSBY-ELSEVIER
T1  - Autoinflammatory mutation in NLRC4 reveals a leucine-rich repeat (LRR)-LRR oligomerization interface
DO  - 10.1016/j.jaci.2018.04.033
IS  - Journal of Allergy and Clinical Immunology
VL  - 142
IS  - 6
SP  - 1956-+
L1  - /bitstream/handle/11343/255350/PMC6281029.pdf?sequence=1&isAllowed=y
ER  -