All missense variants of the NLRP3 gene, listed in Infevers — a comprehensive genetic variant database focused on hereditary autoinflammatory diseases — have undergone testing using the ASC speck assay. This assay is designed to measure the activation of inflammation, which plays a key role in many autoinflammatory conditions. The results of these tests are now publicly available and easily accessible through the Infevers website. This resource is particularly valuable for researchers, clinicians, and geneticists, offering insights into the functional impact of NLRP3 mutations and contributing to a better understanding of their role in disease mechanisms. By providing detailed data on these variants, Infevers supports ongoing efforts to improve diagnostics, treatments, and patient outcomes in the field of autoinflammatory disorders.

The same functional characterization strategy has also been applied to NLRP1 variants, and the corresponding results are available through the Infevers database.

Please consult Infevers for detailed results and variant information.

WP1
A major milestone of the project has been the integration of Eurofever clinical data into the Human Phenotype Ontology (HPO) framework to standardize phenotypic coding in systemic autoinflammatory diseases (SAIDs). In total, 222 Eurofever variables were successfully transcoded into HPO terminology, representing the first large-scale validation of HPO terms in real-world SAID patients.

Using this harmonized dataset, several artificial intelligence and machine-learning approaches were evaluated to improve disease classification and diagnostic support. Among the tested models, the XGBoost algorithm showed the best performance, achieving an average diagnostic accuracy of 80% for major SAIDs including FMF, CAPS, MKD, PFAPA, and TRAPS.

A user-friendly web application has also been developed to support clinicians by providing probability-based SAID diagnosis predictions based on HPO terms.
https://doi.org/10.1016/j.ard.2025.06.438

WP2
Patient enrollment has been completed, including the collection of 70 PBMC samples and 164 DNA samples. Genotyping using the GSA array was successfully performed in 134 Familial Mediterranean Fever (FMF) patients homozygous for the MEFV variants M694V, M694I, or M680I registered in the Eurofever database, as well as in 39 DADA2 patients.

Polygenic Risk Score (PRS), epigenomic, and NEATseq analyses are currently ongoing to investigate the molecular mechanisms underlying disease variability and genotype–phenotype correlations in systemic autoinflammatory diseases (SAIDs).

In parallel, a novel severity score for Deficiency of Adenosine Deaminase 2 (DADA2) has been developed and is currently being validated using real-world patient data from the Eurofever registry.

WP3
WP3 focuses on the functional characterization of variants associated with systemic autoinflammatory diseases (SAIDs) in order to improve variant interpretation and support clinical diagnosis.

Functional characterization results for NLRP3 and NLRP1 variants are already publicly available through the Infevers database, providing accessible resources for the international scientific and clinical community.

In parallel, optimization work for functional assays related to PSTPIP1 and NLRC4 variants is currently ongoing.

Mechanisms of NLRP3 activation and inhibition elucidated by functional analysis of disease-associated variants