{"gene":"NLRC4","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2001,"finding":"NLRC4 (Ipaf) was identified as a human Apaf-1-related protein containing an N-terminal CARD domain, a central nucleotide-binding domain, and a C-terminal LRR domain. Ipaf associates directly and specifically with the CARD domain of procaspase-1 through CARD-CARD interaction. An LRR-domain-deleted constitutively active form of Ipaf induced autocatalytic processing and activation of procaspase-1 and caspase-1-dependent apoptosis in transfected cells.","method":"Co-immunoprecipitation, overexpression in transfected cells, deletion mutagenesis, caspase activation assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct in vitro binding assay with mutagenesis, reconstituted caspase-1 activation, replicated by subsequent studies","pmids":["11390368"],"is_preprint":false},{"year":2004,"finding":"Ipaf (NLRC4)-deficient macrophages fail to activate caspase-1 in response to intracellular Salmonella typhimurium but retain ATP-driven, TLR-stimulated caspase-1 activation (which requires ASC). Cell death triggered by caspase-1-engaging stimuli was ablated in macrophages lacking Ipaf, establishing Ipaf as providing a specific conduit to the inflammasome for intracellular pathogen signals.","method":"Knockout macrophages (Ipaf-/-, ASC-/-, RIP2-/- mice), caspase-1 activation assays, IL-1β/IL-18 maturation, cell death assays, Salmonella infection","journal":"Nature","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean KO with defined cellular phenotype, multiple stimuli and cytokine readouts, independently replicated","pmids":["15190255"],"is_preprint":false},{"year":2006,"finding":"Cytosolic bacterial flagellin activates caspase-1 and IL-1β secretion through NLRC4 (Ipaf) independently of TLR5. Introduction of purified flagellin directly into the macrophage cytoplasm recapitulates Ipaf-dependent caspase-1 activation; a functional SPI1 type III secretion system is required to deliver flagellin to the cytosol.","method":"Ipaf-/- macrophages, cytoplasmic flagellin microinjection/delivery, caspase-1 activation and IL-1β secretion assays, flagellin mutant bacteria, TLR5 independence confirmed","journal":"Nature immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — two independent simultaneous studies (PMIDs 16648853 and 16648852) with KO macrophages and direct ligand delivery","pmids":["16648853","16648852"],"is_preprint":false},{"year":2006,"finding":"Ipaf (NLRC4) restricts Legionella pneumophila replication in mouse macrophages by regulating phagosome maturation: in the absence of Ipaf or caspase-1 activation, the Legionella-containing phagosome acquires ER-derived vesicles and avoids lysosomal fusion, permitting bacterial replication. Flagellin and a competent type IV secretion system are required to activate caspase-1 through Ipaf.","method":"Ipaf-/- and caspase-1-/- macrophages, intracellular bacterial replication assays, phagosome maturation tracking, flagellin-deficient Legionella mutants, in vivo mouse infection","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — KO macrophages with defined phagosome maturation phenotype plus in vivo confirmation","pmids":["16984919"],"is_preprint":false},{"year":2007,"finding":"NLRC4 (Ipaf) mediates caspase-1 activation and pyroptosis in Shigella-infected macrophages independently of flagellin (unlike Salmonella/Legionella). Ipaf is critical for pyroptosis while ASC is dispensable for cell death; absence of Ipaf or caspase-1 (but not ASC) markedly increases autophagy, revealing that Ipaf/caspase-1 suppresses autophagy during Shigella infection.","method":"Ipaf-/- and ASC-/- macrophages, caspase-1 activation assays, cell death (pyroptosis) measurement, autophagy quantification, 3-methyladenine inhibitor, flagellin-deficient Shigella","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — KO macrophages with multiple orthogonal readouts (caspase-1, cell death, autophagy), pharmacological validation","pmids":["17696608"],"is_preprint":false},{"year":2007,"finding":"NLRC4 (IPAF) is required for caspase-1 activation and IL-1β release in macrophages infected with Pseudomonas aeruginosa in a flagellin-independent manner. IPAF- or caspase-1-deficient macrophages are markedly resistant to P. aeruginosa-induced cell death. The P. aeruginosa effector ExoU inhibits IPAF-dependent caspase-1-driven cytokine production.","method":"IPAF-/- and caspase-1-/- macrophages, P. aeruginosa infection, caspase-1 activation, IL-1β release, cell death assays, ExoU-expressing bacterial strains","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — KO macrophages with defined phenotype, orthogonal readouts, confirmed by independent study (PMID 17935074)","pmids":["18070936","17935074"],"is_preprint":false},{"year":2009,"finding":"Caspase-7 is activated downstream of the NLRC4 inflammasome in a caspase-1-dependent manner during Legionella pneumophila infection of murine macrophages; this requires flagellin and functional Naip5. Caspase-7-deficient mice and macrophages are permissive to L. pneumophila replication due to defective lysosomal delivery and delayed cell death.","method":"Nlrc4-/-, caspase-1-/-, caspase-7-/- macrophages and mice, L. pneumophila infection, bacterial replication assays, lysosome fusion tracking","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple KO models with defined bacterial replication and phagosome phenotype","pmids":["19343209"],"is_preprint":false},{"year":2011,"finding":"NAIP5 is a universal component of the flagellin-NLRC4 pathway: NAIP5 directly and specifically binds flagellin, and upon engagement promotes a physical NAIP5-NLRC4 association, enabling full reconstitution of a flagellin-responsive NLRC4 inflammasome in non-macrophage cells. NAIP2 functions analogously as a specific inflammasome receptor for T3SS rod proteins (e.g., PrgJ). Human NAIP (sole human NAIP) recognizes the T3SS needle protein CprI.","method":"Direct binding assays (NAIP5-flagellin), co-immunoprecipitation (NAIP5-NLRC4), reconstitution in non-macrophage cells, NAIP2/NAIP5 specificity assays with multiple bacterial ligands, genetic analysis with C. violaceum infection","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstituted NLRC4 inflammasome in non-macrophage cells, direct binding assays, multiple orthogonal methods, replicated","pmids":["21918512"],"is_preprint":false},{"year":2012,"finding":"NLRC4 is phosphorylated on a single conserved residue Ser533 by PKCδ following Salmonella infection of macrophages. Phosphorylation of Ser533 is critical for NLRC4 inflammasome function: NLRC4 S533A fails to activate caspase-1, pyroptosis, or recruit procaspase-1, and does not assemble inflammasome specks. Phosphomimetic S533D causes rapid pyroptosis without infection. PKCδ phosphorylates NLRC4 S533 in vitro, and Prkcd-/- macrophages show attenuated caspase-1 activation specifically in response to S. typhimurium.","method":"Knock-in mice (NLRC4-3xFlag), phospho-Ser533 antibody, reconstitution with S533A/S533D mutants in Nlrc4-/- macrophages, in vitro kinase assay with recombinant PKCδ, immunodepletion of PKCδ from lysates, Prkcd-/- macrophages","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro kinase assay plus mutagenesis plus KO validation, multiple orthogonal methods in one rigorous study","pmids":["22885697"],"is_preprint":false},{"year":2013,"finding":"Crystal structure of mouse NLRC4 reveals its autoinhibition mechanism: ADP-mediated interaction between the NBD and WHD stabilizes the closed conformation; HD2 repressively contacts a conserved α-helix of the NBD; the LRR domain sterically occludes one face of the NBD to maintain NLRC4 in a monomeric state. Disruption of ADP-mediated NBD-WHD or NBD-HD2/NBD-LRR interactions causes constitutive NLRC4 activation.","method":"X-ray crystallography (crystal structure of mouse NLRC4), site-directed mutagenesis of ADP-binding and domain-interface residues, functional activation assays","journal":"Science","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure with functional mutagenesis validation in a single rigorous study","pmids":["23765277"],"is_preprint":false},{"year":2014,"finding":"A de novo gain-of-function mutation in NLRC4 (p.Val341Ala) in the HD1 domain causes constitutive IL-1 family cytokine production, macrophage cell death, and an autoinflammatory syndrome. Macrophages from affected individuals show constitutive inflammasome component aggregation and pyroptosis.","method":"Whole-exome sequencing, in vitro functional assays in patient macrophages (cytokine production, cell death, inflammasome staining), cosegregation analysis","journal":"Nature genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — patient macrophage functional assays with multiple orthogonal readouts, gain-of-function mechanism directly tested","pmids":["25217960"],"is_preprint":false},{"year":2014,"finding":"A missense mutation in NLRC4 promotes formation of NLRC4-containing inflammasomes that cleave procaspase-1 and increase IL-1β production, causing a familial cold autoinflammatory syndrome. Transgenic mice expressing mutant Nlrc4 develop dermatitis and arthritis dependent on IL-1β-mediated IL-17A production from neutrophils.","method":"Patient genetic analysis, in vitro inflammasome activation assays, transgenic mouse model, IL-1β/IL-17A cytokine measurements","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — transgenic mouse model plus in vitro inflammasome reconstitution, multiple orthogonal readouts","pmids":["25385754"],"is_preprint":false},{"year":2015,"finding":"Cryo-EM structure of a PrgJ-NAIP2-NLRC4 complex at 6.6 Å reveals a wheel-like architecture in which NLRC4 activation involves substantial structural reorganization creating one oligomerization (catalytic) surface. Once activated, NLRC4 uses this catalytic surface to activate an inactive NLRC4 monomer, self-propagating its active conformation. NAIP proteins possess a catalytic surface matching only the receptor surface of NLRC4, ensuring one NAIP initiates NLRC4 oligomerization.","method":"Cryo-EM structure determination (6.6 Å resolution), biochemical reconstitution of PrgJ-NAIP2-NLRC4 complex, structural analysis of oligomerization interfaces","journal":"Science","confidence":"High","confidence_rationale":"Tier 1 / Strong — cryo-EM structure with biochemical reconstitution, mechanistic model validated structurally","pmids":["26449475"],"is_preprint":false},{"year":2015,"finding":"NLRC4 Ser533 phosphorylation occurs independently of NAIP5 detection of flagellin (Naip5 deletion abolishes caspase-1 activation but not NLRC4 phosphorylation). H. pylori flagellin triggers NLRC4 Ser533 phosphorylation but fails to activate caspase-1, establishing a biphasic activation mechanism: Ser533 phosphorylation (via the flagellin D0 domain) primes NLRC4 for subsequent activation by the flagellin sensor NAIP5 (which requires the flagellin C-terminus).","method":"Naip5-/- macrophages, Nlrc4-/- macrophages, H. pylori flagellin and Y. enterocolitica infection, flagellin domain deletion mutants, phospho-Ser533 antibody, caspase-1/IL-1β/pyroptosis assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple KO macrophage lines, flagellin domain mutants, multiple orthogonal readouts defining a two-step mechanism","pmids":["25605939"],"is_preprint":false},{"year":2016,"finding":"NLRP3 associates with NLRC4 (via the NLRC4 NACHT domain) in macrophages infected with Salmonella typhimurium or transfected with flagellin. When NLRC4 Ser533 phosphorylation is absent (S533A), NLRC4 can still activate caspase-1 by recruiting NLRP3 and its adaptor ASC, revealing an unexpected functional overlap between the two inflammasome scaffolds.","method":"Co-immunoprecipitation (NLRC4-NLRP3 association), Nlrc4(S533A/S533A) knock-in BMDMs, Nlrc4(S533A/S533A)/Nlrp3-/- double mutant BMDMs, caspase-1 activation assays","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP plus genetic epistasis with knock-in and double KO mice","pmids":["27139490"],"is_preprint":false},{"year":2018,"finding":"IRF8 transcription factor is required for optimal NLRC4 inflammasome activation specifically in response to flagellin/T3SS-expressing bacteria (Salmonella, Burkholderia, Pseudomonas) but not for NLRP3, AIM2, or Pyrin inflammasomes. IRF8 governs transcription of Naip genes, thereby enabling NAIP-dependent NLRC4 inflammasome activation.","method":"IRF8-deficient BMDMs, bacterial infection assays, caspase-1/IL-1β/pyroptosis readouts, RNA-seq, ChIP (IRF8 binding to Naip promoters), in vivo mouse infection","journal":"Cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — KO macrophages with specific inflammasome selectivity, ChIP confirmation of transcriptional mechanism, in vivo validation","pmids":["29576451"],"is_preprint":false},{"year":2018,"finding":"NLRC4 activates an alternative ASC- and caspase-8-dependent apoptotic cell death program in caspase-1-deficient macrophages (morphologically distinct from pyroptosis). Caspase-1 catalytically dead knock-in macrophages confirm that caspase-1 protease activity suppresses this alternative caspase-8/ASC apoptotic pathway downstream of NLRC4. GSDME/DFNA5 is dispensable for secondary necrosis following NLRC4-mediated apoptosis.","method":"Caspase-1-/- BMDMs, genome-wide CRISPR/Cas9 screen, gene-targeted mice (Casp1 C284A KI, FADD, caspase-8, ASC), cell death morphology, flow cytometry","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — CRISPR screen plus multiple gene-targeted mouse lines with orthogonal validation","pmids":["29491424"],"is_preprint":false},{"year":2018,"finding":"NLRC4 (CARD-based sensor) triggers caspase-8-mediated apoptosis as an alternative cell death in caspase-1-/- macrophages and intestinal epithelial organoids. FADD is recruited to ASC specks which serve as cytosolic platforms for caspase-8 activation. TLR-induced c-FLIP expression inhibits caspase-8-mediated apoptosis downstream of ASC speck assembly without affecting pyroptosis.","method":"Caspase-1-/- macrophages, intestinal epithelial organoids, ASC speck formation assay, co-immunoprecipitation (FADD-ASC specks), c-FLIP overexpression, flow cytometry","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple cell types, reciprocal co-IP, genetic and pharmacological validation of FADD/caspase-8 pathway","pmids":["29262324"],"is_preprint":false},{"year":2010,"finding":"Ipaf (NLRC4) interacts with Sug1 (a component of the 26S proteasome) via amino acids 91-253 of Ipaf (the same region that the LRR domain binds intramolecularly). Sug1 co-expression enables ubiquitination of Ipaf, leading to cytoplasmic aggregate formation, caspase-8 recruitment and activation, and cell death independently of caspase-1. The LRR domain of Ipaf negatively regulates this apoptosis-inducing function by competing with Sug1 binding.","method":"Yeast two-hybrid screen, co-immunoprecipitation (Ipaf-Sug1), co-localization imaging, ubiquitination assays, RNAi, dominant-negative approaches, caspase activation assays","journal":"The Biochemical journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — yeast two-hybrid plus co-IP plus ubiquitination assay, single lab","pmids":["20085538"],"is_preprint":false},{"year":2005,"finding":"Ipaf (NLRC4) is a p53-inducible gene: p53 directly induces Ipaf transcription via a functional p53-binding site in the Ipaf promoter. A dominant-negative Ipaf mutant and Ipaf-directed shRNA both inhibit p53-dependent and doxorubicin-induced apoptosis by ~50%.","method":"Reporter gene assay (p53-binding site in Ipaf promoter), p53 overexpression, doxorubicin treatment of p53-positive and p53-negative cell lines, dominant-negative Ipaf, shRNA knockdown","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reporter assay plus RNAi plus dominant-negative, single lab, multiple cell lines","pmids":["15580302"],"is_preprint":false},{"year":2006,"finding":"Ipaf (NLRC4) activated by caspase-1 causes Bax activation at mitochondria, leading to mitochondrial membrane permeabilization and release of cytochrome c and Omi, functioning as an initiator upstream of mitochondrial events. Bcl2 inhibits mitochondrial permeabilization induced downstream of Ipaf/caspase-1 but not caspase-1 activation itself.","method":"Dominant-negative Ipaf, shRNA, caspase-1 overexpression, Bcl2 overexpression, dominant-negative caspase-9, Bax activation assay, mitochondrial membrane potential assay","journal":"The FEBS journal","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — multiple approaches but no direct biochemical reconstitution; single lab","pmids":["16817903"],"is_preprint":false},{"year":2021,"finding":"DDX17, a DExD/H box RNA helicase, senses endogenous SINE RNAs (retrotransposon-derived short interspersed nuclear elements) and licenses assembly of a non-canonical NLRC4 inflammasome comprising NLRC4, NLRP3, and ASC, inducing caspase-1 activation and cytokine release independently of NAIPs. DDX17 inhibition decreased IL-18 in SLE patient PBMCs and prevented retinal degeneration in an AMD model.","method":"Co-immunoprecipitation (DDX17-NLRC4-NLRP3-ASC complex), DDX17 knockdown/knockout, caspase-1 activation assays, in vivo AMD mouse model, ex vivo SLE patient PBMCs","journal":"Science immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — co-IP of complex plus genetic KD/KO with functional readouts in multiple systems including patient cells and in vivo model","pmids":["34860583"],"is_preprint":false},{"year":2023,"finding":"NLRC4 promotes cGAS-STING-mediated antiviral innate immunity by facilitating CBL (E3 ubiquitin ligase)-mediated K63-linked polyubiquitination of TBK1. NLRC4 enhances the TBK1-CBL interaction; Nlrc4-/- macrophages and mice show attenuated TBK1 phosphorylation, reduced IFN-β/ISG transcription, and increased HSV-1 replication.","method":"NLRC4 overexpression/knockdown/knockout, co-immunoprecipitation (NLRC4-TBK1-CBL), K63-ubiquitination assay, IFN-β promoter reporter, Nlrc4-/- mice with HSV-1 infection","journal":"Journal of medical virology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus ubiquitination assay plus KO mouse in vivo, single lab","pmids":["37537877"],"is_preprint":false},{"year":2021,"finding":"NLRC4 inflammasome-dependent cell death executes through three complementary pathways in a specific order: (1) caspase-1/GSDMD-dependent pyroptosis (default); (2) when pyroptosis is blocked, ASC-mediated caspase-8-dependent apoptosis; (3) when both are blocked, caspase-1-triggered intrinsic apoptosis. Only combined genetic deletion of all three pathways prevents NLRC4-mediated cell death and mouse lethality.","method":"Genetic deletion of caspase-1, GSDMD, ASC, caspase-8, caspase-9 in various combinations; mouse lethality assays; cytokine measurements; Salmonella infection model","journal":"Science advances","confidence":"High","confidence_rationale":"Tier 2 / Strong — combinatorial genetic deletions with in vivo lethality readouts establishing epistatic order of three death pathways","pmids":["34678072"],"is_preprint":false},{"year":2018,"finding":"A gain-of-function mutation in the LRR domain of NLRC4 (p.W655C) causes autoinflammatory disease by engaging an LRR-LRR oligomerization interface. Residues D1010, D1011, L1012, and I1015 on the opposing LRR domain participate in LRR-LRR oligomerization triggered by mutant NLRC4 or PrgI stimulation. ASC contributes to cytokine release but not cell death downstream of this mutation.","method":"CRISPR/Cas9-generated THP-1 cells with WT or W655C NLRC4, ASC speck formation (flow cytometry), cell death and IL-1β/IL-18 ELISA, mutagenesis of LRR interface residues","journal":"The Journal of allergy and clinical immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR KI cells plus site-directed mutagenesis of oligomerization interface, single lab","pmids":["29778503"],"is_preprint":false},{"year":2024,"finding":"Cryo-EM structure of human NAIP/NLRC4 inflammasome assembled with B. thailandensis T3SS needle protein reveals that human NAIP senses T3SS pathogen components via a 'lock-key' activation model involving a lasso-like motif and large structural rearrangement, driving a cascade of NLRC4 protomer assembly to form the full human NLRC4 inflammasome.","method":"Cryo-EM structure determination of human NAIP/NLRC4 complex, biochemical reconstitution with T3SS needle protein, structural analysis","journal":"Nature structural & molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — cryo-EM structure with biochemical reconstitution, providing atomic-level mechanistic detail","pmids":["38177670"],"is_preprint":false},{"year":2023,"finding":"Cryo-EM structure of unliganded mouse NAIP5 reveals an unprecedented wide-open conformation with the nucleating surface fully exposed, accessible to recruit inactive NLRC4. Upon ligand binding, the WHD of NAIP5 undergoes ~20° rotation causing a steric clash with inactive NLRC4, triggering NLRC4 conformational change from inactive to active. The rotated WHD 17-18 loop directly contacts active NLRC4 to stabilize the NAIP5-NLRC4 complex.","method":"Cryo-EM structure of unliganded NAIP5, comparative structural analysis of ligand-bound vs. unbound states, biochemical binding assays","journal":"Nature structural & molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — cryo-EM structure with mechanistic structural analysis of NAIP activation and NLRC4 recruitment","pmids":["36604500"],"is_preprint":false},{"year":2020,"finding":"Serially conducted genetic experiments using a new Nlrc4-/- mouse line, Nlrc4-S533D phosphomimetic, and Nlrc4-S533A non-phosphorylatable mice fail to detect a requirement for Ser533 phosphorylation in NLRC4 inflammasome function in vivo and in vitro. No role for NLRP3 in NLRC4 function was found using these models (contradicting some earlier claims about NLRP3 rescue).","method":"New Nlrc4-/- mouse line, S533D and S533A knock-in mice, caspase-1 activation, IL-1β, pyroptosis assays in vivo and in vitro, melanoma tumor model","journal":"The Journal of experimental medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — rigorous knock-in mouse model, single lab; negative result contradicting prior positive findings, confidence lowered due to contradiction with PMID 22885697","pmids":["32342103"],"is_preprint":false},{"year":2019,"finding":"High glucose (hyperglycemia) induces NLRC4 phosphorylation in macrophages, which activates the NF-κB/Caspase-1 cascade via an IRF8-dependent pathway, driving macrophage senescence and SASP factor secretion. Deletion of NLRC4 or IRF8 abolishes hyperglycemia-induced cellular senescence and SASP. Metformin inhibits NLRC4 phosphorylation and reduces senescence in this context.","method":"NLRC4-/- and IRF8-/- macrophages, diabetic mouse model (gingival tissue), NLRC4 phosphorylation assay, NF-κB/caspase-1 activation, senescence markers, metformin treatment","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO macrophages plus in vivo diabetic model, IRF8 pathway established, single lab","pmids":["31676687"],"is_preprint":false},{"year":2021,"finding":"Brd4 forms a complex with IRF8/PU.1 and binds to the promoters of Naip genes to maintain their transcription, thereby enabling NLRC4 inflammasome activation. Brd4-deficient BMDMs show impaired caspase-1 activation, ASC oligomerization, IL-1β maturation, GSDMD cleavage, and pyroptosis in response to Salmonella; Brd4 myeloid conditional KO mice show increased susceptibility.","method":"Brd4 conditional KO BMDMs and mice, RNA-seq, RT-PCR, ChIP (Brd4/IRF8/PU.1 at Naip promoters), caspase-1/IL-1β/pyroptosis assays, in vivo Salmonella infection","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — ChIP confirms mechanistic model, KO macrophages and in vivo validation, multiple orthogonal methods","pmids":["33535228"],"is_preprint":false},{"year":2016,"finding":"The tick salivary protein sialostatin L2 binds annexin A2, which impairs NLRC4 inflammasome oligomerization and caspase-1 activation. Annexin A2-deficient macrophages show defective NLRC4 inflammasome oligomerization and reduced IL-1β/IL-18 secretion during Anaplasma phagocytophilum infection. Annexin a2-/- mice are more susceptible to A. phagocytophilum infection.","method":"Sialostatin L2 binding assay to annexin A2, annexin A2-/- macrophages, NLRC4 oligomerization assay, caspase-1 activation, IL-1β/IL-18 ELISA, in vivo mouse infection","journal":"Infection and immunity","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO macrophages plus direct binding assay plus in vivo, single lab, identifies annexin A2 as required for NLRC4 oligomerization","pmids":["27045038"],"is_preprint":false},{"year":2021,"finding":"NLRC4, NLRP3, AIM2, and Pyrin form a large multi-sensor PANoptosome complex (together with ASC, caspase-1, caspase-8, and RIPK3) when cells are exposed to multiple simultaneous PAMPs/DAMPs, driving PANoptosis (combined pyroptosis/apoptosis/necroptosis). This multiprotein complex can be released extracellularly and induce inflammation in neighboring macrophages upon engulfment.","method":"Co-immunoprecipitation of multi-sensor complex, CRISPR knockout of individual sensors, cell death assays (pyroptosis/apoptosis/necroptosis), extracellular complex characterization, macrophage engulfment assay","journal":"Cellular & molecular immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus CRISPR KO, single lab, novel multi-sensor complex finding","pmids":["38008850"],"is_preprint":false}],"current_model":"NLRC4 is a cytosolic NLR-family adaptor that forms the NAIP-NLRC4 inflammasome: NAIP proteins (NAIP2, NAIP5/6 in mice; single NAIP in humans) directly bind bacterial flagellin or type III/IV secretion system components, engage NLRC4 via a structural rearrangement that exposes NLRC4's catalytic oligomerization surface, and trigger ADP-to-ATP exchange-driven NLRC4 oligomerization into a wheel-like complex that recruits and activates procaspase-1; active caspase-1 cleaves GSDMD, pro-IL-1β, and pro-IL-18 to drive pyroptosis and inflammation, with PKCδ-mediated phosphorylation of NLRC4 Ser533 proposed (though contested) as a priming step, and alternative ASC/caspase-8-dependent apoptosis activated when pyroptosis is blocked; gain-of-function mutations in NLRC4 cause constitutive inflammasome activation and autoinflammatory disease in humans."},"narrative":{"mechanistic_narrative":"NLRC4 is a cytosolic NLR-family CARD-containing adaptor that nucleates an inflammasome to detect intracellular bacterial infection and drive caspase-1-dependent inflammation and cell death [PMID:11390368, PMID:15190255]. It contains an N-terminal CARD that binds procaspase-1 directly through CARD-CARD interaction, a central nucleotide-binding domain, and a C-terminal LRR domain that sterically autoinhibits the protein in an ADP-stabilized closed monomeric state [PMID:11390368, PMID:23765277]. Activation is initiated by NAIP receptor proteins, which directly bind bacterial flagellin (NAIP5) or type III secretion system rod/needle proteins (NAIP2, human NAIP), then engage NLRC4 and trigger a large conformational rearrangement that exposes a catalytic oligomerization surface; activated NLRC4 propagates its active conformation to recruit successive inactive monomers, building a wheel-like inflammasome that recruits and activates procaspase-1 [PMID:21918512, PMID:26449475, PMID:38177670, PMID:36604500]. Through this pathway NLRC4 controls caspase-1 activation, IL-1β/IL-18 maturation, and pyroptosis in response to Salmonella, Legionella, Shigella, and Pseudomonas, restricting bacterial replication by regulating phagosome maturation and downstream caspase-7 activation [PMID:15190255, PMID:16648853, PMID:16648852, PMID:16984919, PMID:18070936, PMID:17935074, PMID:19343209]. NLRC4-driven death proceeds through an ordered hierarchy: GSDMD-dependent pyroptosis by default, ASC/caspase-8-dependent apoptosis when pyroptosis is blocked, and caspase-1-triggered intrinsic apoptosis when both are unavailable [PMID:29491424, PMID:29262324, PMID:34678072]. NAIP gene expression is transcriptionally licensed by IRF8 and an IRF8/PU.1/Brd4 complex, setting the threshold for inflammasome activation [PMID:29576451, PMID:33535228]. Gain-of-function mutations in NLRC4 — including HD1 p.V341A and LRR p.W655C — cause constitutive inflammasome assembly, pyroptosis, and human autoinflammatory disease [PMID:25217960, PMID:25385754, PMID:29778503].","teleology":[{"year":2001,"claim":"Established NLRC4 as a caspase-1-engaging adaptor, defining the molecular basis by which it could couple upstream signals to caspase activation.","evidence":"Co-IP and deletion mutagenesis in transfected cells showing CARD-CARD binding to procaspase-1 and caspase-1 activation by an LRR-deleted constitutively active form","pmids":["11390368"],"confidence":"High","gaps":["Did not identify the physiological upstream activator","Overexpression-based, not reconstituted with a defined ligand"]},{"year":2004,"claim":"Demonstrated a non-redundant physiological role: NLRC4 is the specific conduit linking intracellular bacterial sensing to caspase-1, distinct from ASC-dependent TLR signals.","evidence":"Ipaf-/- macrophages with Salmonella infection and caspase-1/cytokine/cell death readouts","pmids":["15190255"],"confidence":"High","gaps":["The bacterial ligand sensed was not defined","Mechanism of NLRC4 engagement unknown"]},{"year":2006,"claim":"Identified the activating ligand and its delivery route, showing cytosolic flagellin delivered via a secretion system activates NLRC4 independently of TLR5.","evidence":"Cytoplasmic flagellin delivery and flagellin-mutant bacteria in Ipaf-/- macrophages (two simultaneous studies)","pmids":["16648853","16648852"],"confidence":"High","gaps":["Direct flagellin-NLRC4 binding was not shown","Did not explain flagellin-independent activation by other pathogens"]},{"year":2006,"claim":"Extended NLRC4 function to antibacterial defense, showing it restricts intracellular replication by controlling phagosome maturation.","evidence":"Ipaf-/- and caspase-1-/- macrophages with Legionella replication and phagosome maturation tracking, plus in vivo infection","pmids":["16984919"],"confidence":"High","gaps":["Molecular link between caspase-1 and phagosome-lysosome fusion not resolved here"]},{"year":2007,"claim":"Revealed pathogen-specific wiring and a link to autophagy, showing NLRC4 drives pyroptosis even when flagellin is dispensable and suppresses autophagy.","evidence":"Ipaf-/- and ASC-/- macrophages infected with Shigella and Pseudomonas, with cell death and autophagy readouts","pmids":["17696608","18070936","17935074"],"confidence":"High","gaps":["Identity of the flagellin-independent activating signal unresolved","Mechanism of autophagy suppression undefined"]},{"year":2009,"claim":"Defined a downstream effector, placing caspase-7 activation under NLRC4/caspase-1 control to mediate bacterial restriction.","evidence":"Nlrc4-/-, caspase-1-/-, caspase-7-/- macrophages and mice with Legionella replication and lysosome fusion assays","pmids":["19343209"],"confidence":"High","gaps":["Caspase-7 substrates in this context not defined"]},{"year":2011,"claim":"Solved how NLRC4 acquires ligand specificity, establishing NAIP proteins as the direct ligand receptors that hand off to NLRC4.","evidence":"Direct NAIP5-flagellin binding, NAIP5-NLRC4 co-IP, and reconstitution in non-macrophage cells with NAIP2/NAIP-ligand specificity","pmids":["21918512"],"confidence":"High","gaps":["Structural basis of NAIP-induced NLRC4 conformational change not yet visualized","Stoichiometry of the complex unknown"]},{"year":2013,"claim":"Defined the autoinhibition mechanism structurally, explaining how NLRC4 is held monomeric until activation.","evidence":"X-ray crystal structure of mouse NLRC4 with mutagenesis of ADP-binding and domain-interface residues","pmids":["23765277"],"confidence":"High","gaps":["Captured only the closed monomer, not the active oligomer","Did not show NAIP engagement"]},{"year":2015,"claim":"Visualized the active inflammasome, revealing self-propagating wheel-like oligomerization seeded by a single NAIP.","evidence":"Cryo-EM of reconstituted PrgJ-NAIP2-NLRC4 complex at 6.6 Å with oligomerization-interface analysis","pmids":["26449475"],"confidence":"High","gaps":["Resolution limited atomic-level interpretation","Human complex not structurally resolved"]},{"year":2024,"claim":"Provided atomic-level mechanism for human inflammasome assembly via a lock-key NAIP activation model.","evidence":"Cryo-EM of human NAIP/NLRC4 with T3SS needle protein; plus unliganded NAIP5 cryo-EM defining the conformational trigger","pmids":["38177670","36604500"],"confidence":"High","gaps":["Dynamics of protomer recruitment not directly observed","How priming modifications affect this transition unresolved"]},{"year":2012,"claim":"Proposed a priming step, identifying PKCδ phosphorylation of NLRC4 Ser533 as required for inflammasome function.","evidence":"Knock-in mice, in vitro PKCδ kinase assay, S533A/S533D reconstitution, and Prkcd-/- macrophages","pmids":["22885697"],"confidence":"High","gaps":["Whether phosphorylation is universally required was later contested","Relationship to NAIP engagement initially unclear"]},{"year":2015,"claim":"Reconciled phosphorylation with ligand sensing as a biphasic model: Ser533 priming via flagellin D0 precedes NAIP5 sensing of the flagellin C-terminus.","evidence":"Naip5-/- and Nlrc4-/- macrophages, flagellin domain mutants, and H. pylori vs Y. enterocolitica infection with phospho-Ser533 readout","pmids":["25605939"],"confidence":"High","gaps":["Kinase responsible in all settings not fully resolved","In vivo requirement remained debated"]},{"year":2020,"claim":"Challenged the Ser533 priming requirement, finding no need for the phosphorylation in vivo with new genetic models.","evidence":"New Nlrc4-/-, S533D, and S533A knock-in mice with caspase-1/IL-1β/pyroptosis assays and melanoma model","pmids":["32342103"],"confidence":"Medium","gaps":["Negative result contradicting prior positive findings (PMID 22885697)","Strain or model differences not fully resolved","Single lab"]},{"year":2016,"claim":"Uncovered functional crosstalk between inflammasome scaffolds, showing NLRC4 can recruit NLRP3/ASC.","evidence":"Reciprocal co-IP of NLRC4-NLRP3 and genetic epistasis with S533A knock-in and Nlrp3-/- BMDMs","pmids":["27139490"],"confidence":"High","gaps":["NLRP3 requirement contested by later work (PMID 32342103)","Physiological contribution of crosstalk unclear"]},{"year":2018,"claim":"Identified transcriptional licensing of the pathway, with IRF8 controlling Naip gene expression to enable NLRC4 activation selectively for bacterial sensors.","evidence":"IRF8-/- BMDMs, RNA-seq, ChIP at Naip promoters, and in vivo bacterial infection","pmids":["29576451"],"confidence":"High","gaps":["Signals controlling IRF8 activity in this context not defined"]},{"year":2021,"claim":"Extended transcriptional control, showing Brd4 cooperates with IRF8/PU.1 to maintain Naip transcription.","evidence":"Brd4 conditional KO BMDMs/mice, ChIP of Brd4/IRF8/PU.1 at Naip promoters, and Salmonella infection","pmids":["33535228"],"confidence":"High","gaps":["Whether Brd4 has direct inflammasome roles beyond transcription unknown"]},{"year":2018,"claim":"Defined alternative death programs, establishing that NLRC4 drives ASC/caspase-8 apoptosis when caspase-1/pyroptosis is unavailable.","evidence":"Caspase-1-/- macrophages and organoids, CRISPR screen, FADD/caspase-8/ASC genetic models, and c-FLIP regulation","pmids":["29491424","29262324"],"confidence":"High","gaps":["In vivo relevance of the apoptotic branch quantified later"]},{"year":2021,"claim":"Established the full hierarchy of NLRC4-driven death as three ordered, complementary pathways.","evidence":"Combinatorial deletion of caspase-1, GSDMD, ASC, caspase-8, caspase-9 with mouse lethality and Salmonella infection","pmids":["34678072"],"confidence":"High","gaps":["Tissue-specific contributions of each branch not dissected"]},{"year":2014,"claim":"Connected NLRC4 to human disease, showing gain-of-function mutations cause constitutive inflammasome activation and autoinflammation.","evidence":"Exome sequencing, patient macrophage functional assays (V341A), and transgenic mouse modeling of an autoinflammatory mutation","pmids":["25217960","25385754"],"confidence":"High","gaps":["Genotype-phenotype correlation across mutations incomplete"]},{"year":2018,"claim":"Defined a structural mechanism for one disease mutation, implicating an LRR-LRR oligomerization interface.","evidence":"CRISPR knock-in THP-1 cells with W655C and mutagenesis of opposing LRR interface residues","pmids":["29778503"],"confidence":"Medium","gaps":["Single lab; LRR-LRR interface model not corroborated structurally here","Generalizability to other mutations unclear"]},{"year":2021,"claim":"Identified a NAIP-independent NLRC4 inflammasome triggered by endogenous RNA, broadening the activation repertoire toward sterile/autoimmune contexts.","evidence":"Co-IP of DDX17-NLRC4-NLRP3-ASC complex, DDX17 KD/KO with functional readouts, SLE PBMCs, and AMD model","pmids":["34860583"],"confidence":"High","gaps":["How DDX17 engages NLRC4 without NAIP undefined","Structural basis of the non-canonical complex unknown"]},{"year":null,"claim":"How NLRC4 priming, scaffold crosstalk (NLRP3), and non-canonical/NAIP-independent activation are integrated under physiological conditions remains unresolved, with the Ser533 and NLRP3 requirements directly contested.","evidence":"Conflicting genetic findings across labs (PMID 22885697/27139490 vs 32342103)","pmids":[],"confidence":"Medium","gaps":["Reconciliation of contradictory Ser533/NLRP3 requirements","Physiological scope of non-canonical DDX17 and PANoptosome complexes","Mechanism of NAIP-independent assembly"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,1]},{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[9,12]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[2,7]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,7]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,2,7]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[1,2,7]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[4,16,23]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[10,11,24]}],"complexes":["NAIP-NLRC4 inflammasome","NLRC4-NLRP3-ASC non-canonical inflammasome","PANoptosome"],"partners":["CASP1","NAIP5","NAIP2","NLRP3","PYCARD","DDX17","PRKCD","TBK1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NPP4","full_name":"NLR family CARD domain-containing protein 4","aliases":["CARD, LRR, and NACHT-containing protein","CED-4-like protein Clan","Caspase recruitment domain-containing protein 12","Ice protease-activating factor","Ipaf"],"length_aa":1024,"mass_kda":116.2,"function":"Key component of inflammasomes that indirectly senses specific proteins from pathogenic bacteria and fungi and responds by assembling an inflammasome complex that promotes caspase-1 activation, cytokine production and macrophage pyroptosis (PubMed:15107016). The NLRC4 inflammasome is activated as part of the innate immune response to a range of intracellular bacteria (By similarity)","subcellular_location":"Cytoplasm; Cytoplasm, cytosol; Inflammasome","url":"https://www.uniprot.org/uniprotkb/Q9NPP4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/NLRC4","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/NLRC4","total_profiled":1310},"omim":[{"mim_id":"616115","title":"FAMILIAL COLD AUTOINFLAMMATORY SYNDROME 4; FCAS4","url":"https://www.omim.org/entry/616115"},{"mim_id":"616050","title":"AUTOINFLAMMATION WITH INFANTILE ENTEROCOLITIS; AIFEC","url":"https://www.omim.org/entry/616050"},{"mim_id":"607585","title":"ATM SERINE/THREONINE KINASE; ATM","url":"https://www.omim.org/entry/607585"},{"mim_id":"606848","title":"NIMA-RELATED KINASE 7; NEK7","url":"https://www.omim.org/entry/606848"},{"mim_id":"606831","title":"NLR FAMILY, CASPASE RECRUITMENT DOMAIN-CONTAINING 4; NLRC4","url":"https://www.omim.org/entry/606831"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Cytosol","reliability":"Supported"},{"location":"Vesicles","reliability":"Additional"},{"location":"Plasma membrane","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":8.5},{"tissue":"lymphoid tissue","ntpm":11.3}],"url":"https://www.proteinatlas.org/search/NLRC4"},"hgnc":{"alias_symbol":["CLAN1","ipaf","CLANA","CLANB","CLANC","CLAND","CLR2.1","CLAN"],"prev_symbol":["CARD12"]},"alphafold":{"accession":"Q9NPP4","domains":[{"cath_id":"1.10.533.10","chopping":"2-85","consensus_level":"high","plddt":84.8302,"start":2,"end":85},{"cath_id":"3.40.50.300","chopping":"118-299","consensus_level":"high","plddt":86.9125,"start":118,"end":299},{"cath_id":"1.10.1900.50","chopping":"302-358","consensus_level":"medium","plddt":90.3646,"start":302,"end":358},{"cath_id":"3.80.10.10","chopping":"919-1024","consensus_level":"medium","plddt":87.1016,"start":919,"end":1024}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NPP4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NPP4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NPP4-F1-predicted_aligned_error_v6.png","plddt_mean":85.12},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NLRC4","jax_strain_url":"https://www.jax.org/strain/search?query=NLRC4"},"sequence":{"accession":"Q9NPP4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NPP4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NPP4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NPP4"}},"corpus_meta":[{"pmid":"15190255","id":"PMC_15190255","title":"Differential activation of the inflammasome by caspase-1 adaptors ASC and Ipaf.","date":"2004","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/15190255","citation_count":1483,"is_preprint":false},{"pmid":"21918512","id":"PMC_21918512","title":"The NLRC4 inflammasome receptors for bacterial flagellin and type III secretion apparatus.","date":"2011","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/21918512","citation_count":1065,"is_preprint":false},{"pmid":"16648853","id":"PMC_16648853","title":"Cytoplasmic flagellin activates caspase-1 and secretion of interleukin 1beta via Ipaf.","date":"2006","source":"Nature immunology","url":"https://pubmed.ncbi.nlm.nih.gov/16648853","citation_count":932,"is_preprint":false},{"pmid":"16648852","id":"PMC_16648852","title":"Cytosolic flagellin requires Ipaf for activation of caspase-1 and interleukin 1beta in salmonella-infected macrophages.","date":"2006","source":"Nature immunology","url":"https://pubmed.ncbi.nlm.nih.gov/16648852","citation_count":931,"is_preprint":false},{"pmid":"17696608","id":"PMC_17696608","title":"Differential regulation of caspase-1 activation, pyroptosis, and autophagy via Ipaf and ASC in Shigella-infected macrophages.","date":"2007","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/17696608","citation_count":433,"is_preprint":false},{"pmid":"18070936","id":"PMC_18070936","title":"Immune recognition of Pseudomonas aeruginosa mediated by the IPAF/NLRC4 inflammasome.","date":"2007","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/18070936","citation_count":425,"is_preprint":false},{"pmid":"25217960","id":"PMC_25217960","title":"Mutation of NLRC4 causes a syndrome of enterocolitis and autoinflammation.","date":"2014","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/25217960","citation_count":395,"is_preprint":false},{"pmid":"11390368","id":"PMC_11390368","title":"Identification of Ipaf, a human caspase-1-activating protein related to Apaf-1.","date":"2001","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11390368","citation_count":380,"is_preprint":false},{"pmid":"16984919","id":"PMC_16984919","title":"Regulation of Legionella phagosome maturation and infection through flagellin and host Ipaf.","date":"2006","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16984919","citation_count":379,"is_preprint":false},{"pmid":"28404595","id":"PMC_28404595","title":"NLR members NLRC4 and NLRP3 mediate sterile inflammasome activation in microglia and astrocytes.","date":"2017","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/28404595","citation_count":340,"is_preprint":false},{"pmid":"23765277","id":"PMC_23765277","title":"Crystal structure of NLRC4 reveals its autoinhibition mechanism.","date":"2013","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/23765277","citation_count":325,"is_preprint":false},{"pmid":"26449475","id":"PMC_26449475","title":"Structural and biochemical basis for induced self-propagation of NLRC4.","date":"2015","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/26449475","citation_count":290,"is_preprint":false},{"pmid":"22885697","id":"PMC_22885697","title":"Phosphorylation of NLRC4 is critical for inflammasome activation.","date":"2012","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/22885697","citation_count":254,"is_preprint":false},{"pmid":"20490635","id":"PMC_20490635","title":"Involvement of the AIM2, NLRC4, and NLRP3 inflammasomes in caspase-1 activation by Listeria monocytogenes.","date":"2010","source":"Journal of clinical immunology","url":"https://pubmed.ncbi.nlm.nih.gov/20490635","citation_count":233,"is_preprint":false},{"pmid":"25775556","id":"PMC_25775556","title":"AIM2 and NLRC4 inflammasomes contribute with ASC to acute brain injury independently of NLRP3.","date":"2015","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/25775556","citation_count":231,"is_preprint":false},{"pmid":"17690885","id":"PMC_17690885","title":"TLR5 and Ipaf: dual sensors of bacterial flagellin in the innate immune system.","date":"2007","source":"Seminars in immunopathology","url":"https://pubmed.ncbi.nlm.nih.gov/17690885","citation_count":228,"is_preprint":false},{"pmid":"25385754","id":"PMC_25385754","title":"An inherited mutation in NLRC4 causes autoinflammation in human and mice.","date":"2014","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/25385754","citation_count":212,"is_preprint":false},{"pmid":"17935074","id":"PMC_17935074","title":"Critical role for Ipaf in Pseudomonas aeruginosa-induced caspase-1 activation.","date":"2007","source":"European journal of immunology","url":"https://pubmed.ncbi.nlm.nih.gov/17935074","citation_count":211,"is_preprint":false},{"pmid":"27708283","id":"PMC_27708283","title":"Obesity-associated NLRC4 inflammasome activation drives breast cancer progression.","date":"2016","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/27708283","citation_count":198,"is_preprint":false},{"pmid":"25395539","id":"PMC_25395539","title":"Viral infection. Prevention and cure of rotavirus infection via TLR5/NLRC4-mediated production of IL-22 and IL-18.","date":"2014","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/25395539","citation_count":182,"is_preprint":false},{"pmid":"25879286","id":"PMC_25879286","title":"The NAIP-NLRC4 inflammasome in innate immune detection of bacterial flagellin and type III secretion apparatus.","date":"2015","source":"Immunological reviews","url":"https://pubmed.ncbi.nlm.nih.gov/25879286","citation_count":173,"is_preprint":false},{"pmid":"19343209","id":"PMC_19343209","title":"Caspase-7 activation by the Nlrc4/Ipaf inflammasome restricts Legionella pneumophila infection.","date":"2009","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/19343209","citation_count":165,"is_preprint":false},{"pmid":"25621709","id":"PMC_25621709","title":"The NAIP/NLRC4 inflammasomes.","date":"2015","source":"Current opinion in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/25621709","citation_count":161,"is_preprint":false},{"pmid":"29576451","id":"PMC_29576451","title":"IRF8 Regulates Transcription of Naips for NLRC4 Inflammasome Activation.","date":"2018","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/29576451","citation_count":160,"is_preprint":false},{"pmid":"30195027","id":"PMC_30195027","title":"Evidence that NLRC4 inflammasome mediates apoptotic and pyroptotic microglial death following ischemic stroke.","date":"2018","source":"Brain, behavior, and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/30195027","citation_count":158,"is_preprint":false},{"pmid":"19237518","id":"PMC_19237518","title":"Asc and Ipaf Inflammasomes direct distinct pathways for caspase-1 activation in response to Legionella pneumophila.","date":"2009","source":"Infection and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/19237518","citation_count":157,"is_preprint":false},{"pmid":"32295623","id":"PMC_32295623","title":"NLRP12 collaborates with NLRP3 and NLRC4 to promote pyroptosis inducing ganglion cell death of acute glaucoma.","date":"2020","source":"Molecular neurodegeneration","url":"https://pubmed.ncbi.nlm.nih.gov/32295623","citation_count":139,"is_preprint":false},{"pmid":"38008850","id":"PMC_38008850","title":"Integrated NLRP3, AIM2, NLRC4, Pyrin inflammasome activation and assembly drive PANoptosis.","date":"2023","source":"Cellular & molecular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/38008850","citation_count":137,"is_preprint":false},{"pmid":"24280936","id":"PMC_24280936","title":"NLRC4 expression in intestinal epithelial cells mediates protection against an enteric pathogen.","date":"2013","source":"Mucosal immunology","url":"https://pubmed.ncbi.nlm.nih.gov/24280936","citation_count":127,"is_preprint":false},{"pmid":"27139490","id":"PMC_27139490","title":"NLRP3 recruitment by NLRC4 during Salmonella infection.","date":"2016","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/27139490","citation_count":124,"is_preprint":false},{"pmid":"29491424","id":"PMC_29491424","title":"ASC- and caspase-8-dependent apoptotic pathway diverges from the NLRC4 inflammasome in macrophages.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/29491424","citation_count":121,"is_preprint":false},{"pmid":"29262324","id":"PMC_29262324","title":"Caspase-1 Engagement and TLR-Induced c-FLIP Expression Suppress ASC/Caspase-8-Dependent Apoptosis by Inflammasome Sensors NLRP1b and NLRC4.","date":"2017","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/29262324","citation_count":121,"is_preprint":false},{"pmid":"22318495","id":"PMC_22318495","title":"Cytosolic flagellin receptor NLRC4 protects mice against mucosal and systemic challenges.","date":"2012","source":"Mucosal immunology","url":"https://pubmed.ncbi.nlm.nih.gov/22318495","citation_count":114,"is_preprint":false},{"pmid":"21072873","id":"PMC_21072873","title":"TLR5 or NLRC4 is necessary and sufficient for promotion of humoral immunity by flagellin.","date":"2010","source":"European journal of immunology","url":"https://pubmed.ncbi.nlm.nih.gov/21072873","citation_count":110,"is_preprint":false},{"pmid":"18453601","id":"PMC_18453601","title":"NAIP and Ipaf control Legionella pneumophila replication in human cells.","date":"2008","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/18453601","citation_count":106,"is_preprint":false},{"pmid":"33494299","id":"PMC_33494299","title":"Advances in Understanding Activation and Function of the NLRC4 Inflammasome.","date":"2021","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33494299","citation_count":104,"is_preprint":false},{"pmid":"28957823","id":"PMC_28957823","title":"NLRC4 inflammasomopathies.","date":"2017","source":"Current opinion in allergy and clinical immunology","url":"https://pubmed.ncbi.nlm.nih.gov/28957823","citation_count":103,"is_preprint":false},{"pmid":"25605939","id":"PMC_25605939","title":"Flagellin-induced NLRC4 phosphorylation primes the inflammasome for activation by NAIP5.","date":"2015","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/25605939","citation_count":102,"is_preprint":false},{"pmid":"33074100","id":"PMC_33074100","title":"NAIP-NLRC4-deficient mice are susceptible to shigellosis.","date":"2020","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/33074100","citation_count":85,"is_preprint":false},{"pmid":"33380493","id":"PMC_33380493","title":"Salmonella Flagellin Activates NAIP/NLRC4 and Canonical NLRP3 Inflammasomes in Human Macrophages.","date":"2020","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/33380493","citation_count":85,"is_preprint":false},{"pmid":"32499055","id":"PMC_32499055","title":"The NAIP/NLRC4 inflammasome in infection and pathology.","date":"2020","source":"Molecular aspects of medicine","url":"https://pubmed.ncbi.nlm.nih.gov/32499055","citation_count":75,"is_preprint":false},{"pmid":"32729154","id":"PMC_32729154","title":"Molecular mechanisms activating the NAIP-NLRC4 inflammasome: Implications in infectious disease, autoinflammation, and cancer.","date":"2020","source":"Immunological reviews","url":"https://pubmed.ncbi.nlm.nih.gov/32729154","citation_count":75,"is_preprint":false},{"pmid":"27617861","id":"PMC_27617861","title":"NLRC4 suppresses melanoma tumor progression independently of inflammasome activation.","date":"2016","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/27617861","citation_count":70,"is_preprint":false},{"pmid":"30658261","id":"PMC_30658261","title":"Variants in NLRP3 and NLRC4 inflammasome associate with susceptibility and severity of multiple sclerosis.","date":"2019","source":"Multiple sclerosis and related disorders","url":"https://pubmed.ncbi.nlm.nih.gov/30658261","citation_count":66,"is_preprint":false},{"pmid":"35073381","id":"PMC_35073381","title":"Human NAIP/NLRC4 and NLRP3 inflammasomes detect Salmonella type III secretion system activities to restrict intracellular bacterial replication.","date":"2022","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/35073381","citation_count":66,"is_preprint":false},{"pmid":"18819842","id":"PMC_18819842","title":"NLRC4/IPAF: a CARD carrying member of the NLR family.","date":"2008","source":"Clinical immunology (Orlando, Fla.)","url":"https://pubmed.ncbi.nlm.nih.gov/18819842","citation_count":64,"is_preprint":false},{"pmid":"31676687","id":"PMC_31676687","title":"Hyperglycemia-induced inflamm-aging accelerates gingival senescence via NLRC4 phosphorylation.","date":"2019","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/31676687","citation_count":63,"is_preprint":false},{"pmid":"33858325","id":"PMC_33858325","title":"LncRNA-Fendrr protects against the ubiquitination and degradation of NLRC4 protein through HERC2 to regulate the pyroptosis of microglia.","date":"2021","source":"Molecular medicine (Cambridge, Mass.)","url":"https://pubmed.ncbi.nlm.nih.gov/33858325","citation_count":61,"is_preprint":false},{"pmid":"35678562","id":"PMC_35678562","title":"Salmonella enterica Serovar Typhimurium Induces NAIP/NLRC4- and NLRP3/ASC-Independent, Caspase-4-Dependent Inflammasome Activation in Human Intestinal Epithelial Cells.","date":"2022","source":"Infection and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/35678562","citation_count":58,"is_preprint":false},{"pmid":"34276697","id":"PMC_34276697","title":"Updating the NLRC4 Inflammasome: from Bacterial Infections to Autoimmunity and Cancer.","date":"2021","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/34276697","citation_count":56,"is_preprint":false},{"pmid":"29778503","id":"PMC_29778503","title":"Autoinflammatory mutation in NLRC4 reveals a leucine-rich repeat (LRR)-LRR oligomerization interface.","date":"2018","source":"The Journal of allergy and clinical immunology","url":"https://pubmed.ncbi.nlm.nih.gov/29778503","citation_count":55,"is_preprint":false},{"pmid":"25071770","id":"PMC_25071770","title":"Emerging Concepts about NAIP/NLRC4 Inflammasomes.","date":"2014","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/25071770","citation_count":53,"is_preprint":false},{"pmid":"32342103","id":"PMC_32342103","title":"NLRC4 inflammasome activation is NLRP3- and phosphorylation-independent during infection and does not protect from melanoma.","date":"2020","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/32342103","citation_count":52,"is_preprint":false},{"pmid":"15580302","id":"PMC_15580302","title":"Caspase-1 activator Ipaf is a p53-inducible gene involved in apoptosis.","date":"2005","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/15580302","citation_count":52,"is_preprint":false},{"pmid":"17382568","id":"PMC_17382568","title":"ASC, Ipaf and Cryopyrin/Nalp3: bona fide intracellular adapters of the caspase-1 inflammasome.","date":"2007","source":"Microbes and infection","url":"https://pubmed.ncbi.nlm.nih.gov/17382568","citation_count":51,"is_preprint":false},{"pmid":"28963531","id":"PMC_28963531","title":"Methylene blue inhibits NLRP3, NLRC4, AIM2, and non-canonical inflammasome activation.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28963531","citation_count":51,"is_preprint":false},{"pmid":"34678072","id":"PMC_34678072","title":"NLRC4 inflammasome-dependent cell death occurs by a complementary series of three death pathways and determines lethality in mice.","date":"2021","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/34678072","citation_count":50,"is_preprint":false},{"pmid":"34860583","id":"PMC_34860583","title":"DDX17 is an essential mediator of sterile NLRC4 inflammasome activation by retrotransposon RNAs.","date":"2021","source":"Science immunology","url":"https://pubmed.ncbi.nlm.nih.gov/34860583","citation_count":47,"is_preprint":false},{"pmid":"24337744","id":"PMC_24337744","title":"Arsenic trioxide and other arsenical compounds inhibit the NLRP1, NLRP3, and NAIP5/NLRC4 inflammasomes.","date":"2013","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/24337744","citation_count":47,"is_preprint":false},{"pmid":"27706238","id":"PMC_27706238","title":"Involvement of the NLRC4-Inflammasome in Diabetic Nephropathy.","date":"2016","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/27706238","citation_count":46,"is_preprint":false},{"pmid":"30368901","id":"PMC_30368901","title":"The Salmonella pathogenicity island-2 subverts human NLRP3 and NLRC4 inflammasome responses.","date":"2018","source":"Journal of leukocyte biology","url":"https://pubmed.ncbi.nlm.nih.gov/30368901","citation_count":43,"is_preprint":false},{"pmid":"27045038","id":"PMC_27045038","title":"The Tick Protein Sialostatin L2 Binds to Annexin A2 and Inhibits NLRC4-Mediated Inflammasome Activation.","date":"2016","source":"Infection and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/27045038","citation_count":41,"is_preprint":false},{"pmid":"20085538","id":"PMC_20085538","title":"Interaction with Sug1 enables Ipaf ubiquitination leading to caspase 8 activation and cell death.","date":"2010","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/20085538","citation_count":37,"is_preprint":false},{"pmid":"30824190","id":"PMC_30824190","title":"NLRC4 inflammasome activation regulated by TNF-α promotes inflammatory responses in nonalcoholic fatty liver disease.","date":"2019","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/30824190","citation_count":37,"is_preprint":false},{"pmid":"28137500","id":"PMC_28137500","title":"Interstitial lung disease and myositis-specific and associated autoantibodies: Clinical manifestations, survival and the performance of the new ATS/ERS criteria for interstitial pneumonia with autoimmune features (IPAF).","date":"2016","source":"Respiratory medicine","url":"https://pubmed.ncbi.nlm.nih.gov/28137500","citation_count":37,"is_preprint":false},{"pmid":"31715312","id":"PMC_31715312","title":"Involvement of NLRC4 inflammasome through caspase-1 and IL-1β augments neuroinflammation and contributes to memory impairment in an experimental model of Alzheimer's like disease.","date":"2019","source":"Brain research bulletin","url":"https://pubmed.ncbi.nlm.nih.gov/31715312","citation_count":37,"is_preprint":false},{"pmid":"35120997","id":"PMC_35120997","title":"LCN2 Mediates Skin Inflammation in Psoriasis through the SREBP2‒NLRC4 Axis.","date":"2022","source":"The Journal of investigative dermatology","url":"https://pubmed.ncbi.nlm.nih.gov/35120997","citation_count":36,"is_preprint":false},{"pmid":"34848837","id":"PMC_34848837","title":"The pivotal role of the NLRC4 inflammasome in neuroinflammation after intracerebral hemorrhage in rats.","date":"2021","source":"Experimental & molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34848837","citation_count":36,"is_preprint":false},{"pmid":"31511355","id":"PMC_31511355","title":"Helicobacter pylori Exploits the NLRC4 Inflammasome to Dampen Host Defenses.","date":"2019","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/31511355","citation_count":36,"is_preprint":false},{"pmid":"32397236","id":"PMC_32397236","title":"NLRC4, ASC and Caspase-1 Are Inflammasome Components that Are Mediated by P2Y2R Activation in Breast Cancer Cells.","date":"2020","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32397236","citation_count":34,"is_preprint":false},{"pmid":"37058933","id":"PMC_37058933","title":"Human and mouse NAIP/NLRC4 inflammasome responses to bacterial infection.","date":"2023","source":"Current opinion in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/37058933","citation_count":33,"is_preprint":false},{"pmid":"25914934","id":"PMC_25914934","title":"Activation of NLRC4 downregulates TLR5-mediated antibody immune responses against flagellin.","date":"2015","source":"Cellular & molecular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/25914934","citation_count":33,"is_preprint":false},{"pmid":"35435150","id":"PMC_35435150","title":"Curcumin activates NLRC4, AIM2, and IFI16 inflammasomes and induces pyroptosis by up-regulated ISG3 transcript factor in acute myeloid leukemia cell lines.","date":"2022","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/35435150","citation_count":33,"is_preprint":false},{"pmid":"35428651","id":"PMC_35428651","title":"Low-ratio somatic NLRC4 mutation causes late-onset autoinflammatory disease.","date":"2022","source":"Annals of the rheumatic diseases","url":"https://pubmed.ncbi.nlm.nih.gov/35428651","citation_count":32,"is_preprint":false},{"pmid":"33535228","id":"PMC_33535228","title":"Brd4 regulates NLRC4 inflammasome activation by facilitating IRF8-mediated transcription of Naips.","date":"2021","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/33535228","citation_count":32,"is_preprint":false},{"pmid":"37537877","id":"PMC_37537877","title":"NLRC4 promotes the cGAS-STING signaling pathway by facilitating CBL-mediated K63-linked polyubiquitination of TBK1.","date":"2023","source":"Journal of medical virology","url":"https://pubmed.ncbi.nlm.nih.gov/37537877","citation_count":31,"is_preprint":false},{"pmid":"37586642","id":"PMC_37586642","title":"Targeting Desulfovibrio vulgaris flagellin-induced NAIP/NLRC4 inflammasome activation in macrophages attenuates ulcerative colitis.","date":"2023","source":"Journal of advanced research","url":"https://pubmed.ncbi.nlm.nih.gov/37586642","citation_count":30,"is_preprint":false},{"pmid":"32531834","id":"PMC_32531834","title":"NLRC4 biology in immunity and inflammation.","date":"2020","source":"Journal of leukocyte biology","url":"https://pubmed.ncbi.nlm.nih.gov/32531834","citation_count":30,"is_preprint":false},{"pmid":"22426978","id":"PMC_22426978","title":"Sensing bacterial infections by NAIP receptors in NLRC4 inflammasome activation.","date":"2012","source":"Protein & cell","url":"https://pubmed.ncbi.nlm.nih.gov/22426978","citation_count":29,"is_preprint":false},{"pmid":"35922848","id":"PMC_35922848","title":"Aprepitant attenuates NLRC4-dependent neuronal pyroptosis via NK1R/PKCδ pathway in a mouse model of intracerebral hemorrhage.","date":"2022","source":"Journal of neuroinflammation","url":"https://pubmed.ncbi.nlm.nih.gov/35922848","citation_count":29,"is_preprint":false},{"pmid":"34783940","id":"PMC_34783940","title":"Recessive NLRC4-Autoinflammatory Disease Reveals an Ulcerative Colitis Locus.","date":"2021","source":"Journal of clinical immunology","url":"https://pubmed.ncbi.nlm.nih.gov/34783940","citation_count":28,"is_preprint":false},{"pmid":"29721174","id":"PMC_29721174","title":"Epigenetic hypomethylation and upregulation of NLRC4 and NLRP12 in Kawasaki disease.","date":"2018","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/29721174","citation_count":28,"is_preprint":false},{"pmid":"31551961","id":"PMC_31551961","title":"Involvement of NLRP3 and NLRC4 Inflammasome in Uropathogenic E. coli Mediated Urinary Tract Infections.","date":"2019","source":"Frontiers in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/31551961","citation_count":28,"is_preprint":false},{"pmid":"25402425","id":"PMC_25402425","title":"Redundant and cooperative interactions between TLR5 and NLRC4 in protective lung mucosal immunity against Pseudomonas aeruginosa.","date":"2014","source":"Journal of innate immunity","url":"https://pubmed.ncbi.nlm.nih.gov/25402425","citation_count":26,"is_preprint":false},{"pmid":"31870725","id":"PMC_31870725","title":"A novel de novo NLRC4 mutation reinforces the likely pathogenicity of specific LRR domain mutation.","date":"2019","source":"Clinical immunology (Orlando, Fla.)","url":"https://pubmed.ncbi.nlm.nih.gov/31870725","citation_count":26,"is_preprint":false},{"pmid":"34898657","id":"PMC_34898657","title":"A unique NLRC4 receptor from echinoderms mediates Vibrio phagocytosis via rearrangement of the cytoskeleton and polymerization of F-actin.","date":"2021","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/34898657","citation_count":25,"is_preprint":false},{"pmid":"36604500","id":"PMC_36604500","title":"Mechanism of NAIP-NLRC4 inflammasome activation revealed by cryo-EM structure of unliganded NAIP5.","date":"2023","source":"Nature structural & molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/36604500","citation_count":24,"is_preprint":false},{"pmid":"35835791","id":"PMC_35835791","title":"Circ_0000181 regulates miR-667-5p/NLRC4 axis to promote pyroptosis progression in diabetic nephropathy.","date":"2022","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/35835791","citation_count":23,"is_preprint":false},{"pmid":"35216164","id":"PMC_35216164","title":"Association of Tim-3/Gal-9 Axis with NLRC4 Inflammasome in Glioma Malignancy: Tim-3/Gal-9 Induce the NLRC4 Inflammasome.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35216164","citation_count":23,"is_preprint":false},{"pmid":"26362438","id":"PMC_26362438","title":"Molecular Characterization of the NLRC4 Expression in Relation to Interleukin-18 Levels.","date":"2015","source":"Circulation. Cardiovascular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/26362438","citation_count":22,"is_preprint":false},{"pmid":"37741453","id":"PMC_37741453","title":"Toxoplasma gondii Induces Pyroptosis in Human Placental Trophoblast and Amniotic Cells by Inducing ROS Production and Activation of Cathepsin B and NLRP1/NLRP3/NLRC4/AIM2 Inflammasome.","date":"2023","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/37741453","citation_count":21,"is_preprint":false},{"pmid":"39557108","id":"PMC_39557108","title":"Eriocitrin ameliorates hepatic fibrosis and inflammation: The involvement of PPARα-mediated NLRP1/NLRC4 inflammasome signaling cascades.","date":"2024","source":"Journal of ethnopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/39557108","citation_count":21,"is_preprint":false},{"pmid":"38177670","id":"PMC_38177670","title":"Structural basis of the human NAIP/NLRC4 inflammasome assembly and pathogen sensing.","date":"2024","source":"Nature structural & molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/38177670","citation_count":21,"is_preprint":false},{"pmid":"34335562","id":"PMC_34335562","title":"Salmonella spvC Gene Inhibits Autophagy of Host Cells and Suppresses NLRP3 as Well as NLRC4.","date":"2021","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/34335562","citation_count":21,"is_preprint":false},{"pmid":"32276737","id":"PMC_32276737","title":"Inflammasome activation by NLRP1 and NLRC4 in patients with coronary stenosis.","date":"2020","source":"Immunobiology","url":"https://pubmed.ncbi.nlm.nih.gov/32276737","citation_count":21,"is_preprint":false},{"pmid":"35301258","id":"PMC_35301258","title":"NLRC4 Deficiency Leads to Enhanced Phosphorylation of MLKL and Necroptosis.","date":"2022","source":"ImmunoHorizons","url":"https://pubmed.ncbi.nlm.nih.gov/35301258","citation_count":20,"is_preprint":false},{"pmid":"16817903","id":"PMC_16817903","title":"Involvement of caspase 1 and its activator Ipaf upstream of mitochondrial events in apoptosis.","date":"2006","source":"The FEBS journal","url":"https://pubmed.ncbi.nlm.nih.gov/16817903","citation_count":20,"is_preprint":false},{"pmid":"29617808","id":"PMC_29617808","title":"Novel aspects of the assembly and activation of inflammasomes with focus on the NLRC4 inflammasome.","date":"2018","source":"International immunology","url":"https://pubmed.ncbi.nlm.nih.gov/29617808","citation_count":20,"is_preprint":false},{"pmid":"27958346","id":"PMC_27958346","title":"Clinical Association of Chemokine (C-X-C motif) Ligand 1 (CXCL1) with Interstitial Pneumonia with Autoimmune Features (IPAF).","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/27958346","citation_count":20,"is_preprint":false},{"pmid":"33781745","id":"PMC_33781745","title":"Discovery and characterization of small-molecule inhibitors of NLRP3 and NLRC4 inflammasomes.","date":"2021","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/33781745","citation_count":19,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":51071,"output_tokens":9311,"usd":0.146439,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":19178,"output_tokens":5733,"usd":0.119608,"stage2_stop_reason":"end_turn"},"total_usd":0.266047,"stage1_batch_id":"msgbatch_013a8chtv3AWyTqv4z4sEF4C","stage2_batch_id":"msgbatch_01AHsuW63vinsvuTgzmQ3n62","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2001,\n      \"finding\": \"NLRC4 (Ipaf) was identified as a human Apaf-1-related protein containing an N-terminal CARD domain, a central nucleotide-binding domain, and a C-terminal LRR domain. Ipaf associates directly and specifically with the CARD domain of procaspase-1 through CARD-CARD interaction. An LRR-domain-deleted constitutively active form of Ipaf induced autocatalytic processing and activation of procaspase-1 and caspase-1-dependent apoptosis in transfected cells.\",\n      \"method\": \"Co-immunoprecipitation, overexpression in transfected cells, deletion mutagenesis, caspase activation assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — direct in vitro binding assay with mutagenesis, reconstituted caspase-1 activation, replicated by subsequent studies\",\n      \"pmids\": [\"11390368\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Ipaf (NLRC4)-deficient macrophages fail to activate caspase-1 in response to intracellular Salmonella typhimurium but retain ATP-driven, TLR-stimulated caspase-1 activation (which requires ASC). Cell death triggered by caspase-1-engaging stimuli was ablated in macrophages lacking Ipaf, establishing Ipaf as providing a specific conduit to the inflammasome for intracellular pathogen signals.\",\n      \"method\": \"Knockout macrophages (Ipaf-/-, ASC-/-, RIP2-/- mice), caspase-1 activation assays, IL-1β/IL-18 maturation, cell death assays, Salmonella infection\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean KO with defined cellular phenotype, multiple stimuli and cytokine readouts, independently replicated\",\n      \"pmids\": [\"15190255\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Cytosolic bacterial flagellin activates caspase-1 and IL-1β secretion through NLRC4 (Ipaf) independently of TLR5. Introduction of purified flagellin directly into the macrophage cytoplasm recapitulates Ipaf-dependent caspase-1 activation; a functional SPI1 type III secretion system is required to deliver flagellin to the cytosol.\",\n      \"method\": \"Ipaf-/- macrophages, cytoplasmic flagellin microinjection/delivery, caspase-1 activation and IL-1β secretion assays, flagellin mutant bacteria, TLR5 independence confirmed\",\n      \"journal\": \"Nature immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — two independent simultaneous studies (PMIDs 16648853 and 16648852) with KO macrophages and direct ligand delivery\",\n      \"pmids\": [\"16648853\", \"16648852\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Ipaf (NLRC4) restricts Legionella pneumophila replication in mouse macrophages by regulating phagosome maturation: in the absence of Ipaf or caspase-1 activation, the Legionella-containing phagosome acquires ER-derived vesicles and avoids lysosomal fusion, permitting bacterial replication. Flagellin and a competent type IV secretion system are required to activate caspase-1 through Ipaf.\",\n      \"method\": \"Ipaf-/- and caspase-1-/- macrophages, intracellular bacterial replication assays, phagosome maturation tracking, flagellin-deficient Legionella mutants, in vivo mouse infection\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — KO macrophages with defined phagosome maturation phenotype plus in vivo confirmation\",\n      \"pmids\": [\"16984919\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"NLRC4 (Ipaf) mediates caspase-1 activation and pyroptosis in Shigella-infected macrophages independently of flagellin (unlike Salmonella/Legionella). Ipaf is critical for pyroptosis while ASC is dispensable for cell death; absence of Ipaf or caspase-1 (but not ASC) markedly increases autophagy, revealing that Ipaf/caspase-1 suppresses autophagy during Shigella infection.\",\n      \"method\": \"Ipaf-/- and ASC-/- macrophages, caspase-1 activation assays, cell death (pyroptosis) measurement, autophagy quantification, 3-methyladenine inhibitor, flagellin-deficient Shigella\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — KO macrophages with multiple orthogonal readouts (caspase-1, cell death, autophagy), pharmacological validation\",\n      \"pmids\": [\"17696608\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"NLRC4 (IPAF) is required for caspase-1 activation and IL-1β release in macrophages infected with Pseudomonas aeruginosa in a flagellin-independent manner. IPAF- or caspase-1-deficient macrophages are markedly resistant to P. aeruginosa-induced cell death. The P. aeruginosa effector ExoU inhibits IPAF-dependent caspase-1-driven cytokine production.\",\n      \"method\": \"IPAF-/- and caspase-1-/- macrophages, P. aeruginosa infection, caspase-1 activation, IL-1β release, cell death assays, ExoU-expressing bacterial strains\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — KO macrophages with defined phenotype, orthogonal readouts, confirmed by independent study (PMID 17935074)\",\n      \"pmids\": [\"18070936\", \"17935074\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Caspase-7 is activated downstream of the NLRC4 inflammasome in a caspase-1-dependent manner during Legionella pneumophila infection of murine macrophages; this requires flagellin and functional Naip5. Caspase-7-deficient mice and macrophages are permissive to L. pneumophila replication due to defective lysosomal delivery and delayed cell death.\",\n      \"method\": \"Nlrc4-/-, caspase-1-/-, caspase-7-/- macrophages and mice, L. pneumophila infection, bacterial replication assays, lysosome fusion tracking\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple KO models with defined bacterial replication and phagosome phenotype\",\n      \"pmids\": [\"19343209\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"NAIP5 is a universal component of the flagellin-NLRC4 pathway: NAIP5 directly and specifically binds flagellin, and upon engagement promotes a physical NAIP5-NLRC4 association, enabling full reconstitution of a flagellin-responsive NLRC4 inflammasome in non-macrophage cells. NAIP2 functions analogously as a specific inflammasome receptor for T3SS rod proteins (e.g., PrgJ). Human NAIP (sole human NAIP) recognizes the T3SS needle protein CprI.\",\n      \"method\": \"Direct binding assays (NAIP5-flagellin), co-immunoprecipitation (NAIP5-NLRC4), reconstitution in non-macrophage cells, NAIP2/NAIP5 specificity assays with multiple bacterial ligands, genetic analysis with C. violaceum infection\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstituted NLRC4 inflammasome in non-macrophage cells, direct binding assays, multiple orthogonal methods, replicated\",\n      \"pmids\": [\"21918512\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"NLRC4 is phosphorylated on a single conserved residue Ser533 by PKCδ following Salmonella infection of macrophages. Phosphorylation of Ser533 is critical for NLRC4 inflammasome function: NLRC4 S533A fails to activate caspase-1, pyroptosis, or recruit procaspase-1, and does not assemble inflammasome specks. Phosphomimetic S533D causes rapid pyroptosis without infection. PKCδ phosphorylates NLRC4 S533 in vitro, and Prkcd-/- macrophages show attenuated caspase-1 activation specifically in response to S. typhimurium.\",\n      \"method\": \"Knock-in mice (NLRC4-3xFlag), phospho-Ser533 antibody, reconstitution with S533A/S533D mutants in Nlrc4-/- macrophages, in vitro kinase assay with recombinant PKCδ, immunodepletion of PKCδ from lysates, Prkcd-/- macrophages\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro kinase assay plus mutagenesis plus KO validation, multiple orthogonal methods in one rigorous study\",\n      \"pmids\": [\"22885697\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Crystal structure of mouse NLRC4 reveals its autoinhibition mechanism: ADP-mediated interaction between the NBD and WHD stabilizes the closed conformation; HD2 repressively contacts a conserved α-helix of the NBD; the LRR domain sterically occludes one face of the NBD to maintain NLRC4 in a monomeric state. Disruption of ADP-mediated NBD-WHD or NBD-HD2/NBD-LRR interactions causes constitutive NLRC4 activation.\",\n      \"method\": \"X-ray crystallography (crystal structure of mouse NLRC4), site-directed mutagenesis of ADP-binding and domain-interface residues, functional activation assays\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure with functional mutagenesis validation in a single rigorous study\",\n      \"pmids\": [\"23765277\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"A de novo gain-of-function mutation in NLRC4 (p.Val341Ala) in the HD1 domain causes constitutive IL-1 family cytokine production, macrophage cell death, and an autoinflammatory syndrome. Macrophages from affected individuals show constitutive inflammasome component aggregation and pyroptosis.\",\n      \"method\": \"Whole-exome sequencing, in vitro functional assays in patient macrophages (cytokine production, cell death, inflammasome staining), cosegregation analysis\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — patient macrophage functional assays with multiple orthogonal readouts, gain-of-function mechanism directly tested\",\n      \"pmids\": [\"25217960\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"A missense mutation in NLRC4 promotes formation of NLRC4-containing inflammasomes that cleave procaspase-1 and increase IL-1β production, causing a familial cold autoinflammatory syndrome. Transgenic mice expressing mutant Nlrc4 develop dermatitis and arthritis dependent on IL-1β-mediated IL-17A production from neutrophils.\",\n      \"method\": \"Patient genetic analysis, in vitro inflammasome activation assays, transgenic mouse model, IL-1β/IL-17A cytokine measurements\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — transgenic mouse model plus in vitro inflammasome reconstitution, multiple orthogonal readouts\",\n      \"pmids\": [\"25385754\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Cryo-EM structure of a PrgJ-NAIP2-NLRC4 complex at 6.6 Å reveals a wheel-like architecture in which NLRC4 activation involves substantial structural reorganization creating one oligomerization (catalytic) surface. Once activated, NLRC4 uses this catalytic surface to activate an inactive NLRC4 monomer, self-propagating its active conformation. NAIP proteins possess a catalytic surface matching only the receptor surface of NLRC4, ensuring one NAIP initiates NLRC4 oligomerization.\",\n      \"method\": \"Cryo-EM structure determination (6.6 Å resolution), biochemical reconstitution of PrgJ-NAIP2-NLRC4 complex, structural analysis of oligomerization interfaces\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — cryo-EM structure with biochemical reconstitution, mechanistic model validated structurally\",\n      \"pmids\": [\"26449475\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"NLRC4 Ser533 phosphorylation occurs independently of NAIP5 detection of flagellin (Naip5 deletion abolishes caspase-1 activation but not NLRC4 phosphorylation). H. pylori flagellin triggers NLRC4 Ser533 phosphorylation but fails to activate caspase-1, establishing a biphasic activation mechanism: Ser533 phosphorylation (via the flagellin D0 domain) primes NLRC4 for subsequent activation by the flagellin sensor NAIP5 (which requires the flagellin C-terminus).\",\n      \"method\": \"Naip5-/- macrophages, Nlrc4-/- macrophages, H. pylori flagellin and Y. enterocolitica infection, flagellin domain deletion mutants, phospho-Ser533 antibody, caspase-1/IL-1β/pyroptosis assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple KO macrophage lines, flagellin domain mutants, multiple orthogonal readouts defining a two-step mechanism\",\n      \"pmids\": [\"25605939\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"NLRP3 associates with NLRC4 (via the NLRC4 NACHT domain) in macrophages infected with Salmonella typhimurium or transfected with flagellin. When NLRC4 Ser533 phosphorylation is absent (S533A), NLRC4 can still activate caspase-1 by recruiting NLRP3 and its adaptor ASC, revealing an unexpected functional overlap between the two inflammasome scaffolds.\",\n      \"method\": \"Co-immunoprecipitation (NLRC4-NLRP3 association), Nlrc4(S533A/S533A) knock-in BMDMs, Nlrc4(S533A/S533A)/Nlrp3-/- double mutant BMDMs, caspase-1 activation assays\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP plus genetic epistasis with knock-in and double KO mice\",\n      \"pmids\": [\"27139490\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"IRF8 transcription factor is required for optimal NLRC4 inflammasome activation specifically in response to flagellin/T3SS-expressing bacteria (Salmonella, Burkholderia, Pseudomonas) but not for NLRP3, AIM2, or Pyrin inflammasomes. IRF8 governs transcription of Naip genes, thereby enabling NAIP-dependent NLRC4 inflammasome activation.\",\n      \"method\": \"IRF8-deficient BMDMs, bacterial infection assays, caspase-1/IL-1β/pyroptosis readouts, RNA-seq, ChIP (IRF8 binding to Naip promoters), in vivo mouse infection\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — KO macrophages with specific inflammasome selectivity, ChIP confirmation of transcriptional mechanism, in vivo validation\",\n      \"pmids\": [\"29576451\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"NLRC4 activates an alternative ASC- and caspase-8-dependent apoptotic cell death program in caspase-1-deficient macrophages (morphologically distinct from pyroptosis). Caspase-1 catalytically dead knock-in macrophages confirm that caspase-1 protease activity suppresses this alternative caspase-8/ASC apoptotic pathway downstream of NLRC4. GSDME/DFNA5 is dispensable for secondary necrosis following NLRC4-mediated apoptosis.\",\n      \"method\": \"Caspase-1-/- BMDMs, genome-wide CRISPR/Cas9 screen, gene-targeted mice (Casp1 C284A KI, FADD, caspase-8, ASC), cell death morphology, flow cytometry\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — CRISPR screen plus multiple gene-targeted mouse lines with orthogonal validation\",\n      \"pmids\": [\"29491424\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"NLRC4 (CARD-based sensor) triggers caspase-8-mediated apoptosis as an alternative cell death in caspase-1-/- macrophages and intestinal epithelial organoids. FADD is recruited to ASC specks which serve as cytosolic platforms for caspase-8 activation. TLR-induced c-FLIP expression inhibits caspase-8-mediated apoptosis downstream of ASC speck assembly without affecting pyroptosis.\",\n      \"method\": \"Caspase-1-/- macrophages, intestinal epithelial organoids, ASC speck formation assay, co-immunoprecipitation (FADD-ASC specks), c-FLIP overexpression, flow cytometry\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple cell types, reciprocal co-IP, genetic and pharmacological validation of FADD/caspase-8 pathway\",\n      \"pmids\": [\"29262324\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Ipaf (NLRC4) interacts with Sug1 (a component of the 26S proteasome) via amino acids 91-253 of Ipaf (the same region that the LRR domain binds intramolecularly). Sug1 co-expression enables ubiquitination of Ipaf, leading to cytoplasmic aggregate formation, caspase-8 recruitment and activation, and cell death independently of caspase-1. The LRR domain of Ipaf negatively regulates this apoptosis-inducing function by competing with Sug1 binding.\",\n      \"method\": \"Yeast two-hybrid screen, co-immunoprecipitation (Ipaf-Sug1), co-localization imaging, ubiquitination assays, RNAi, dominant-negative approaches, caspase activation assays\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — yeast two-hybrid plus co-IP plus ubiquitination assay, single lab\",\n      \"pmids\": [\"20085538\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Ipaf (NLRC4) is a p53-inducible gene: p53 directly induces Ipaf transcription via a functional p53-binding site in the Ipaf promoter. A dominant-negative Ipaf mutant and Ipaf-directed shRNA both inhibit p53-dependent and doxorubicin-induced apoptosis by ~50%.\",\n      \"method\": \"Reporter gene assay (p53-binding site in Ipaf promoter), p53 overexpression, doxorubicin treatment of p53-positive and p53-negative cell lines, dominant-negative Ipaf, shRNA knockdown\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reporter assay plus RNAi plus dominant-negative, single lab, multiple cell lines\",\n      \"pmids\": [\"15580302\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Ipaf (NLRC4) activated by caspase-1 causes Bax activation at mitochondria, leading to mitochondrial membrane permeabilization and release of cytochrome c and Omi, functioning as an initiator upstream of mitochondrial events. Bcl2 inhibits mitochondrial permeabilization induced downstream of Ipaf/caspase-1 but not caspase-1 activation itself.\",\n      \"method\": \"Dominant-negative Ipaf, shRNA, caspase-1 overexpression, Bcl2 overexpression, dominant-negative caspase-9, Bax activation assay, mitochondrial membrane potential assay\",\n      \"journal\": \"The FEBS journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — multiple approaches but no direct biochemical reconstitution; single lab\",\n      \"pmids\": [\"16817903\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"DDX17, a DExD/H box RNA helicase, senses endogenous SINE RNAs (retrotransposon-derived short interspersed nuclear elements) and licenses assembly of a non-canonical NLRC4 inflammasome comprising NLRC4, NLRP3, and ASC, inducing caspase-1 activation and cytokine release independently of NAIPs. DDX17 inhibition decreased IL-18 in SLE patient PBMCs and prevented retinal degeneration in an AMD model.\",\n      \"method\": \"Co-immunoprecipitation (DDX17-NLRC4-NLRP3-ASC complex), DDX17 knockdown/knockout, caspase-1 activation assays, in vivo AMD mouse model, ex vivo SLE patient PBMCs\",\n      \"journal\": \"Science immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — co-IP of complex plus genetic KD/KO with functional readouts in multiple systems including patient cells and in vivo model\",\n      \"pmids\": [\"34860583\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"NLRC4 promotes cGAS-STING-mediated antiviral innate immunity by facilitating CBL (E3 ubiquitin ligase)-mediated K63-linked polyubiquitination of TBK1. NLRC4 enhances the TBK1-CBL interaction; Nlrc4-/- macrophages and mice show attenuated TBK1 phosphorylation, reduced IFN-β/ISG transcription, and increased HSV-1 replication.\",\n      \"method\": \"NLRC4 overexpression/knockdown/knockout, co-immunoprecipitation (NLRC4-TBK1-CBL), K63-ubiquitination assay, IFN-β promoter reporter, Nlrc4-/- mice with HSV-1 infection\",\n      \"journal\": \"Journal of medical virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus ubiquitination assay plus KO mouse in vivo, single lab\",\n      \"pmids\": [\"37537877\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NLRC4 inflammasome-dependent cell death executes through three complementary pathways in a specific order: (1) caspase-1/GSDMD-dependent pyroptosis (default); (2) when pyroptosis is blocked, ASC-mediated caspase-8-dependent apoptosis; (3) when both are blocked, caspase-1-triggered intrinsic apoptosis. Only combined genetic deletion of all three pathways prevents NLRC4-mediated cell death and mouse lethality.\",\n      \"method\": \"Genetic deletion of caspase-1, GSDMD, ASC, caspase-8, caspase-9 in various combinations; mouse lethality assays; cytokine measurements; Salmonella infection model\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — combinatorial genetic deletions with in vivo lethality readouts establishing epistatic order of three death pathways\",\n      \"pmids\": [\"34678072\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"A gain-of-function mutation in the LRR domain of NLRC4 (p.W655C) causes autoinflammatory disease by engaging an LRR-LRR oligomerization interface. Residues D1010, D1011, L1012, and I1015 on the opposing LRR domain participate in LRR-LRR oligomerization triggered by mutant NLRC4 or PrgI stimulation. ASC contributes to cytokine release but not cell death downstream of this mutation.\",\n      \"method\": \"CRISPR/Cas9-generated THP-1 cells with WT or W655C NLRC4, ASC speck formation (flow cytometry), cell death and IL-1β/IL-18 ELISA, mutagenesis of LRR interface residues\",\n      \"journal\": \"The Journal of allergy and clinical immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR KI cells plus site-directed mutagenesis of oligomerization interface, single lab\",\n      \"pmids\": [\"29778503\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Cryo-EM structure of human NAIP/NLRC4 inflammasome assembled with B. thailandensis T3SS needle protein reveals that human NAIP senses T3SS pathogen components via a 'lock-key' activation model involving a lasso-like motif and large structural rearrangement, driving a cascade of NLRC4 protomer assembly to form the full human NLRC4 inflammasome.\",\n      \"method\": \"Cryo-EM structure determination of human NAIP/NLRC4 complex, biochemical reconstitution with T3SS needle protein, structural analysis\",\n      \"journal\": \"Nature structural & molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — cryo-EM structure with biochemical reconstitution, providing atomic-level mechanistic detail\",\n      \"pmids\": [\"38177670\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Cryo-EM structure of unliganded mouse NAIP5 reveals an unprecedented wide-open conformation with the nucleating surface fully exposed, accessible to recruit inactive NLRC4. Upon ligand binding, the WHD of NAIP5 undergoes ~20° rotation causing a steric clash with inactive NLRC4, triggering NLRC4 conformational change from inactive to active. The rotated WHD 17-18 loop directly contacts active NLRC4 to stabilize the NAIP5-NLRC4 complex.\",\n      \"method\": \"Cryo-EM structure of unliganded NAIP5, comparative structural analysis of ligand-bound vs. unbound states, biochemical binding assays\",\n      \"journal\": \"Nature structural & molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — cryo-EM structure with mechanistic structural analysis of NAIP activation and NLRC4 recruitment\",\n      \"pmids\": [\"36604500\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Serially conducted genetic experiments using a new Nlrc4-/- mouse line, Nlrc4-S533D phosphomimetic, and Nlrc4-S533A non-phosphorylatable mice fail to detect a requirement for Ser533 phosphorylation in NLRC4 inflammasome function in vivo and in vitro. No role for NLRP3 in NLRC4 function was found using these models (contradicting some earlier claims about NLRP3 rescue).\",\n      \"method\": \"New Nlrc4-/- mouse line, S533D and S533A knock-in mice, caspase-1 activation, IL-1β, pyroptosis assays in vivo and in vitro, melanoma tumor model\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — rigorous knock-in mouse model, single lab; negative result contradicting prior positive findings, confidence lowered due to contradiction with PMID 22885697\",\n      \"pmids\": [\"32342103\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"High glucose (hyperglycemia) induces NLRC4 phosphorylation in macrophages, which activates the NF-κB/Caspase-1 cascade via an IRF8-dependent pathway, driving macrophage senescence and SASP factor secretion. Deletion of NLRC4 or IRF8 abolishes hyperglycemia-induced cellular senescence and SASP. Metformin inhibits NLRC4 phosphorylation and reduces senescence in this context.\",\n      \"method\": \"NLRC4-/- and IRF8-/- macrophages, diabetic mouse model (gingival tissue), NLRC4 phosphorylation assay, NF-κB/caspase-1 activation, senescence markers, metformin treatment\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO macrophages plus in vivo diabetic model, IRF8 pathway established, single lab\",\n      \"pmids\": [\"31676687\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Brd4 forms a complex with IRF8/PU.1 and binds to the promoters of Naip genes to maintain their transcription, thereby enabling NLRC4 inflammasome activation. Brd4-deficient BMDMs show impaired caspase-1 activation, ASC oligomerization, IL-1β maturation, GSDMD cleavage, and pyroptosis in response to Salmonella; Brd4 myeloid conditional KO mice show increased susceptibility.\",\n      \"method\": \"Brd4 conditional KO BMDMs and mice, RNA-seq, RT-PCR, ChIP (Brd4/IRF8/PU.1 at Naip promoters), caspase-1/IL-1β/pyroptosis assays, in vivo Salmonella infection\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — ChIP confirms mechanistic model, KO macrophages and in vivo validation, multiple orthogonal methods\",\n      \"pmids\": [\"33535228\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The tick salivary protein sialostatin L2 binds annexin A2, which impairs NLRC4 inflammasome oligomerization and caspase-1 activation. Annexin A2-deficient macrophages show defective NLRC4 inflammasome oligomerization and reduced IL-1β/IL-18 secretion during Anaplasma phagocytophilum infection. Annexin a2-/- mice are more susceptible to A. phagocytophilum infection.\",\n      \"method\": \"Sialostatin L2 binding assay to annexin A2, annexin A2-/- macrophages, NLRC4 oligomerization assay, caspase-1 activation, IL-1β/IL-18 ELISA, in vivo mouse infection\",\n      \"journal\": \"Infection and immunity\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO macrophages plus direct binding assay plus in vivo, single lab, identifies annexin A2 as required for NLRC4 oligomerization\",\n      \"pmids\": [\"27045038\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NLRC4, NLRP3, AIM2, and Pyrin form a large multi-sensor PANoptosome complex (together with ASC, caspase-1, caspase-8, and RIPK3) when cells are exposed to multiple simultaneous PAMPs/DAMPs, driving PANoptosis (combined pyroptosis/apoptosis/necroptosis). This multiprotein complex can be released extracellularly and induce inflammation in neighboring macrophages upon engulfment.\",\n      \"method\": \"Co-immunoprecipitation of multi-sensor complex, CRISPR knockout of individual sensors, cell death assays (pyroptosis/apoptosis/necroptosis), extracellular complex characterization, macrophage engulfment assay\",\n      \"journal\": \"Cellular & molecular immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus CRISPR KO, single lab, novel multi-sensor complex finding\",\n      \"pmids\": [\"38008850\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NLRC4 is a cytosolic NLR-family adaptor that forms the NAIP-NLRC4 inflammasome: NAIP proteins (NAIP2, NAIP5/6 in mice; single NAIP in humans) directly bind bacterial flagellin or type III/IV secretion system components, engage NLRC4 via a structural rearrangement that exposes NLRC4's catalytic oligomerization surface, and trigger ADP-to-ATP exchange-driven NLRC4 oligomerization into a wheel-like complex that recruits and activates procaspase-1; active caspase-1 cleaves GSDMD, pro-IL-1β, and pro-IL-18 to drive pyroptosis and inflammation, with PKCδ-mediated phosphorylation of NLRC4 Ser533 proposed (though contested) as a priming step, and alternative ASC/caspase-8-dependent apoptosis activated when pyroptosis is blocked; gain-of-function mutations in NLRC4 cause constitutive inflammasome activation and autoinflammatory disease in humans.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"NLRC4 is a cytosolic NLR-family CARD-containing adaptor that nucleates an inflammasome to detect intracellular bacterial infection and drive caspase-1-dependent inflammation and cell death [#0, #1]. It contains an N-terminal CARD that binds procaspase-1 directly through CARD-CARD interaction, a central nucleotide-binding domain, and a C-terminal LRR domain that sterically autoinhibits the protein in an ADP-stabilized closed monomeric state [#0, #9]. Activation is initiated by NAIP receptor proteins, which directly bind bacterial flagellin (NAIP5) or type III secretion system rod/needle proteins (NAIP2, human NAIP), then engage NLRC4 and trigger a large conformational rearrangement that exposes a catalytic oligomerization surface; activated NLRC4 propagates its active conformation to recruit successive inactive monomers, building a wheel-like inflammasome that recruits and activates procaspase-1 [#7, #12, #25, #26]. Through this pathway NLRC4 controls caspase-1 activation, IL-1β/IL-18 maturation, and pyroptosis in response to Salmonella, Legionella, Shigella, and Pseudomonas, restricting bacterial replication by regulating phagosome maturation and downstream caspase-7 activation [#1, #2, #3, #5, #6]. NLRC4-driven death proceeds through an ordered hierarchy: GSDMD-dependent pyroptosis by default, ASC/caspase-8-dependent apoptosis when pyroptosis is blocked, and caspase-1-triggered intrinsic apoptosis when both are unavailable [#16, #17, #23]. NAIP gene expression is transcriptionally licensed by IRF8 and an IRF8/PU.1/Brd4 complex, setting the threshold for inflammasome activation [#15, #29]. Gain-of-function mutations in NLRC4 — including HD1 p.V341A and LRR p.W655C — cause constitutive inflammasome assembly, pyroptosis, and human autoinflammatory disease [#10, #11, #24].\",\n  \"teleology\": [\n    {\n      \"year\": 2001,\n      \"claim\": \"Established NLRC4 as a caspase-1-engaging adaptor, defining the molecular basis by which it could couple upstream signals to caspase activation.\",\n      \"evidence\": \"Co-IP and deletion mutagenesis in transfected cells showing CARD-CARD binding to procaspase-1 and caspase-1 activation by an LRR-deleted constitutively active form\",\n      \"pmids\": [\"11390368\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not identify the physiological upstream activator\", \"Overexpression-based, not reconstituted with a defined ligand\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Demonstrated a non-redundant physiological role: NLRC4 is the specific conduit linking intracellular bacterial sensing to caspase-1, distinct from ASC-dependent TLR signals.\",\n      \"evidence\": \"Ipaf-/- macrophages with Salmonella infection and caspase-1/cytokine/cell death readouts\",\n      \"pmids\": [\"15190255\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The bacterial ligand sensed was not defined\", \"Mechanism of NLRC4 engagement unknown\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Identified the activating ligand and its delivery route, showing cytosolic flagellin delivered via a secretion system activates NLRC4 independently of TLR5.\",\n      \"evidence\": \"Cytoplasmic flagellin delivery and flagellin-mutant bacteria in Ipaf-/- macrophages (two simultaneous studies)\",\n      \"pmids\": [\"16648853\", \"16648852\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct flagellin-NLRC4 binding was not shown\", \"Did not explain flagellin-independent activation by other pathogens\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Extended NLRC4 function to antibacterial defense, showing it restricts intracellular replication by controlling phagosome maturation.\",\n      \"evidence\": \"Ipaf-/- and caspase-1-/- macrophages with Legionella replication and phagosome maturation tracking, plus in vivo infection\",\n      \"pmids\": [\"16984919\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular link between caspase-1 and phagosome-lysosome fusion not resolved here\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Revealed pathogen-specific wiring and a link to autophagy, showing NLRC4 drives pyroptosis even when flagellin is dispensable and suppresses autophagy.\",\n      \"evidence\": \"Ipaf-/- and ASC-/- macrophages infected with Shigella and Pseudomonas, with cell death and autophagy readouts\",\n      \"pmids\": [\"17696608\", \"18070936\", \"17935074\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the flagellin-independent activating signal unresolved\", \"Mechanism of autophagy suppression undefined\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defined a downstream effector, placing caspase-7 activation under NLRC4/caspase-1 control to mediate bacterial restriction.\",\n      \"evidence\": \"Nlrc4-/-, caspase-1-/-, caspase-7-/- macrophages and mice with Legionella replication and lysosome fusion assays\",\n      \"pmids\": [\"19343209\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Caspase-7 substrates in this context not defined\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Solved how NLRC4 acquires ligand specificity, establishing NAIP proteins as the direct ligand receptors that hand off to NLRC4.\",\n      \"evidence\": \"Direct NAIP5-flagellin binding, NAIP5-NLRC4 co-IP, and reconstitution in non-macrophage cells with NAIP2/NAIP-ligand specificity\",\n      \"pmids\": [\"21918512\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of NAIP-induced NLRC4 conformational change not yet visualized\", \"Stoichiometry of the complex unknown\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Defined the autoinhibition mechanism structurally, explaining how NLRC4 is held monomeric until activation.\",\n      \"evidence\": \"X-ray crystal structure of mouse NLRC4 with mutagenesis of ADP-binding and domain-interface residues\",\n      \"pmids\": [\"23765277\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Captured only the closed monomer, not the active oligomer\", \"Did not show NAIP engagement\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Visualized the active inflammasome, revealing self-propagating wheel-like oligomerization seeded by a single NAIP.\",\n      \"evidence\": \"Cryo-EM of reconstituted PrgJ-NAIP2-NLRC4 complex at 6.6 Å with oligomerization-interface analysis\",\n      \"pmids\": [\"26449475\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Resolution limited atomic-level interpretation\", \"Human complex not structurally resolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Provided atomic-level mechanism for human inflammasome assembly via a lock-key NAIP activation model.\",\n      \"evidence\": \"Cryo-EM of human NAIP/NLRC4 with T3SS needle protein; plus unliganded NAIP5 cryo-EM defining the conformational trigger\",\n      \"pmids\": [\"38177670\", \"36604500\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Dynamics of protomer recruitment not directly observed\", \"How priming modifications affect this transition unresolved\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Proposed a priming step, identifying PKCδ phosphorylation of NLRC4 Ser533 as required for inflammasome function.\",\n      \"evidence\": \"Knock-in mice, in vitro PKCδ kinase assay, S533A/S533D reconstitution, and Prkcd-/- macrophages\",\n      \"pmids\": [\"22885697\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether phosphorylation is universally required was later contested\", \"Relationship to NAIP engagement initially unclear\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Reconciled phosphorylation with ligand sensing as a biphasic model: Ser533 priming via flagellin D0 precedes NAIP5 sensing of the flagellin C-terminus.\",\n      \"evidence\": \"Naip5-/- and Nlrc4-/- macrophages, flagellin domain mutants, and H. pylori vs Y. enterocolitica infection with phospho-Ser533 readout\",\n      \"pmids\": [\"25605939\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Kinase responsible in all settings not fully resolved\", \"In vivo requirement remained debated\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Challenged the Ser533 priming requirement, finding no need for the phosphorylation in vivo with new genetic models.\",\n      \"evidence\": \"New Nlrc4-/-, S533D, and S533A knock-in mice with caspase-1/IL-1β/pyroptosis assays and melanoma model\",\n      \"pmids\": [\"32342103\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Negative result contradicting prior positive findings (PMID 22885697)\", \"Strain or model differences not fully resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Uncovered functional crosstalk between inflammasome scaffolds, showing NLRC4 can recruit NLRP3/ASC.\",\n      \"evidence\": \"Reciprocal co-IP of NLRC4-NLRP3 and genetic epistasis with S533A knock-in and Nlrp3-/- BMDMs\",\n      \"pmids\": [\"27139490\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"NLRP3 requirement contested by later work (PMID 32342103)\", \"Physiological contribution of crosstalk unclear\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Identified transcriptional licensing of the pathway, with IRF8 controlling Naip gene expression to enable NLRC4 activation selectively for bacterial sensors.\",\n      \"evidence\": \"IRF8-/- BMDMs, RNA-seq, ChIP at Naip promoters, and in vivo bacterial infection\",\n      \"pmids\": [\"29576451\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Signals controlling IRF8 activity in this context not defined\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Extended transcriptional control, showing Brd4 cooperates with IRF8/PU.1 to maintain Naip transcription.\",\n      \"evidence\": \"Brd4 conditional KO BMDMs/mice, ChIP of Brd4/IRF8/PU.1 at Naip promoters, and Salmonella infection\",\n      \"pmids\": [\"33535228\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether Brd4 has direct inflammasome roles beyond transcription unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Defined alternative death programs, establishing that NLRC4 drives ASC/caspase-8 apoptosis when caspase-1/pyroptosis is unavailable.\",\n      \"evidence\": \"Caspase-1-/- macrophages and organoids, CRISPR screen, FADD/caspase-8/ASC genetic models, and c-FLIP regulation\",\n      \"pmids\": [\"29491424\", \"29262324\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo relevance of the apoptotic branch quantified later\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Established the full hierarchy of NLRC4-driven death as three ordered, complementary pathways.\",\n      \"evidence\": \"Combinatorial deletion of caspase-1, GSDMD, ASC, caspase-8, caspase-9 with mouse lethality and Salmonella infection\",\n      \"pmids\": [\"34678072\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Tissue-specific contributions of each branch not dissected\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Connected NLRC4 to human disease, showing gain-of-function mutations cause constitutive inflammasome activation and autoinflammation.\",\n      \"evidence\": \"Exome sequencing, patient macrophage functional assays (V341A), and transgenic mouse modeling of an autoinflammatory mutation\",\n      \"pmids\": [\"25217960\", \"25385754\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Genotype-phenotype correlation across mutations incomplete\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Defined a structural mechanism for one disease mutation, implicating an LRR-LRR oligomerization interface.\",\n      \"evidence\": \"CRISPR knock-in THP-1 cells with W655C and mutagenesis of opposing LRR interface residues\",\n      \"pmids\": [\"29778503\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab; LRR-LRR interface model not corroborated structurally here\", \"Generalizability to other mutations unclear\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified a NAIP-independent NLRC4 inflammasome triggered by endogenous RNA, broadening the activation repertoire toward sterile/autoimmune contexts.\",\n      \"evidence\": \"Co-IP of DDX17-NLRC4-NLRP3-ASC complex, DDX17 KD/KO with functional readouts, SLE PBMCs, and AMD model\",\n      \"pmids\": [\"34860583\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How DDX17 engages NLRC4 without NAIP undefined\", \"Structural basis of the non-canonical complex unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How NLRC4 priming, scaffold crosstalk (NLRP3), and non-canonical/NAIP-independent activation are integrated under physiological conditions remains unresolved, with the Ser533 and NLRP3 requirements directly contested.\",\n      \"evidence\": \"Conflicting genetic findings across labs (PMID 22885697/27139490 vs 32342103)\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reconciliation of contradictory Ser533/NLRP3 requirements\", \"Physiological scope of non-canonical DDX17 and PANoptosome complexes\", \"Mechanism of NAIP-independent assembly\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [9, 12]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [2, 7]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 7]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 2, 7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [1, 2, 7]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [4, 16, 23]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [10, 11, 24]}\n    ],\n    \"complexes\": [\n      \"NAIP-NLRC4 inflammasome\",\n      \"NLRC4-NLRP3-ASC non-canonical inflammasome\",\n      \"PANoptosome\"\n    ],\n    \"partners\": [\n      \"CASP1\",\n      \"NAIP5\",\n      \"NAIP2\",\n      \"NLRP3\",\n      \"PYCARD\",\n      \"DDX17\",\n      \"PRKCD\",\n      \"TBK1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}