Background and objective Familial Mediterranean fever (FMF) is the most frequent hereditary autoinflammatory disease. Its diagnosis relies on a set of clinical criteria and a genetic confirmation on identification of biallelic pathogenic MEFV variants. MEFV encodes pyrin, an inflammasome sensor. Using a kinase inhibitor, UCN-01, we recently identified that dephosphorylation of FMF-associated pyrin mutants leads to inflammasome activation. The aim of this study was to assess whether quantifying UCN-01-mediated inflammasome activation could discriminate FMF patients from healthy donors (HD) and from patients with other inflammatory disorders (OID).
Methods Real-time pyroptosis and IL-1β secretion were monitored in response to UCN-01 in monocytes from FMF patients (n=67), HD (n=71) and OID patients (n=40). Sensitivity and specificity of the resulting diagnostic tests were determined by receiver operating characteristic curve analyses.
Results Inflammasome monitoring in response to UCN-01 discriminates FMF patients from other individuals. Pyroptosis assessment leads to a fast FMF diagnosis while combining pyroptosis and IL-1β dosage renders UCN-01-based assays highly sensitive and specific. UCN-01-triggered monocytes responses were influenced by MEFV gene dosage and MEFV mutations in a similar way as clinical phenotypes are.
Conclusions UCN-01-based inflammasome assays could be used to rapidly diagnose FMF, with high sensitivity and specificity.
- familial mediterranean fever
Statistics from Altmetric.com
TH and YJ are joint senior authors.
Handling editor Josef S Smolen
Contributors TH and YJ designed the study; FM, TM, AM, MF, TH performed experiments; FM, FA, MR, TH and YJ analysed the data. FM, TH and YJ wrote the manuscript with input from all authors; SG-L, AB, MG-V, PS, AD, AH, SD, GG and YJ included patients, provided clinical information and samples; TH and YJ oversaw the project.
Funding The study (Depist-FMF, registered at clinicaltrials.gov under the unique identifier NCT03747315) was founded by a grant from the Hospices Civils de Lyon (Jeune Chercheur HCL) and a grant from the Agence Nationale de la Recherche/Direction Générale de l’Offre de Soin (FMFgeneToDiag #ANR-17-CE17-0021) and funding from the European Union's Horizon 2020 research and innovation programme under grant agreement #779295 (ImmunAID).
Competing interests YJ, FM, AB, AM and TH are coinventors and owners of a patent 'Methods and kits for diagnosis of familial Mediterranean fever' (WO/2019/048569).
Patient and public involvement Patients and/or the public were involved in the design, or conduct, or reporting or dissemination plans of this research. Refer to the Methods section for further details.
Patient consent for publication Not required.
Ethics approval The study was approved by the French Comité de Protection des Personnes (CPP,#L16‐189) and by the French Comité Consultatif sur le Traitement de l'Information en matière de Recherche dans le domaine de la Santé (CCTIRS, #16.864). The experiments conformed to the principles set out in the WMA Declaration of Helsinki and the Department of Health and Human Services’Belmont Report. The Etablissement Français du Sang provided HD blood in the framework of the convention #14-1820. Informed consent was received from each participant prior to inclusion in the study.
Provenance and peer review Not commissioned; externally peer reviewed.
Data availability statement All data relevant to the study are included in the article or uploaded as supplementary information. Additional data are available upon reasonable request.
If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.