Table 3.
Main genetic functional studies of CAPS-causing mutations.
| Human mutation | Mouse mutation | Human phenotype | Experimental approach | Conditions of activation | Effect of mutation | Author, year/Reference number |
|---|---|---|---|---|---|---|
| NLRP3:c.778C>T; p.(Arg260Trp) | . | MWS | 1.Monocytes isolation and purification2. ELISA and Western Blot using antibody specifically directed against the active fragment (p17) of proIL-1β | LPS | Spontaneous secretion of active IL-1β | Agostini et al., 2004/91 |
| NLRP3:c.778C>T, p.(Arg260Trp); c.907G>A, p.(Asp303Asn); c.1880A>G, p.(Glu627Gly) | . | MWS, CINCA/NOMID, FCAS | 1. Monocytic THP-1 cell line expressing mutant proteins2. NF-kB Assay3. IL-1 ELISA4. Immunoprecipitation5. Small Interfering RNA | ASC-dependent | 1.Constitutive activation of NLRP32. Induction of NF-kB activation3. Increased IL-1β secretion4. Increased ASC binding | Dowds et al., 2004/94 |
| NM_001243133.2(NLRP3):c.1058T>C,p.(Leu353Pro); c.1687T>A, p.(Tyr563Asn); c.1976T>A, p.(Met659Lys) and c.1573G>A, p.(Glu525Lys)/c.592G>A, p.(Val198Met), | . | FCAS | 1.Adherence-enriched monocytes2. quantitative PCR and ELISA | Cold exposure (32°C and 37°C) | 1. Increased cytokine transcription at 16 hours with 32°C of incubation.2.Response to mild hypothermia with an increased IL-1β release and secretion of IL-6 and TNF-α | Rosengren et al., 2007/96 |
| NM_001243133.2(NLRP3):c.1058T>C,p.(Leu353Pro); c.1687T>A, p.(Tyr563Asn) | . | FCAS | 1.Cell lines (HeLa cells) expressing fusion proteins2. IL-1β secretion assay3. Western-blot analysis4. Reverse transcription and real-time PCR5. Ca2+ imaging | Cold exposure | 1.Inflammasome assembly Ca2+- mediated2. Cryo-sensitive foci formation | Karasawa et al., 2022/97 |
| NM_001243133.2(NLRP3):c.907G>A, p.(Asp303Asn); c.1709A>G, p.(Tyr570Cys) | . | CINCA/NOMID | 1.Cell lines (HeLa cells) expressing fusion proteins2. IL-1β secretion assay3. Western-blot analysis4. Reverse transcription and real-time PCR5. Ca2+ imaging | Cold exposure | 1.Inflammasome assembly Ca2+- mediated2. Cryo-sensitive foci formation | Karasawa et al., 2022/97 |
| NM_001243133.2(NLRP3):c.1058T>C; p.Leu353Pro | p.Leu351Pro | FCAS | 1.Conditional knockin;Nlrp3A350VneoR/+ and Nlrp3L351PneoR/+ mice2. In vitro studies: myeloid cells isolation | 1.LPS2.Cold exposure | Hyperactivation of the NLRP3 | Brydges et al., 2009/98 |
| NLRP3:c.778C>T; p.(Arg260Trp) | p.Arg258Trp | MWS | Constitutive knockin | LPS | 1.Increased IL-1β and IL-18 production from APCs2. Contextual increase of Th17 differentitation | Meng et al., 2009/99 |
| NLRP3:c.907G>A; p.(Asp303Asn | . | CINCA/NOMID,MWS | 1. C57BL/6J mice2. HEK293 cells transfection2.Isolation of particles of recombinant ASC and NLRP33.Coimmunoprecipitation and immunoblot analysis4. ELISA5. Multiplex flow cytometry | 1.LPS2.Extracellular caspase-13.Sorrounding macrophage | Increased extracellular release of functional oligomeric inflammasome particles containing both NLRP3 and ASC | Baroja-Mazo et al.,2014/100 |
| NLRP3:c.907G>A; p.(Asp303Asn) | p.Asp301Asn | CINCA/ NOMID |
Conditional knockin; Nlrp3fl(D301N)/+ mouse models; Gsdmd−/− mice | LPS | 1.Constitutive maturation of GSDMD2. Excessive pore formation3. Increased and constitutive production ofIL-1β | Xiao et al., 2018/101 |
CAPS, Cryopyrin associated periodic syndromes; MWS, Muckle Wells Syndrome; LPS, Lipopolysaccharide; ATP, adenosine triphosphate; ASC, apoptosis-associated speck-like protein containing a CARD; IL, interleukin; GSDMD, gasdermin D; CINCA/NOMID, chronic infantile neurologic, cutaneous, and articular syndrome/neonatal-onset multisystem inflammatory disease; FCAS, Familial cold autoinflammatory syndrome; Th, T-helper.
PCR, Polymerase Chain Reaction; ELISA, Enzyme-linked immunosorbent assay; Th17, T helper 17 cells; APCs, Antigen-presenting cells.