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Blood-based test for diagnosis and functional subtyping of familial Mediterranean fever
Hanne Van Gorp1,2, Linyan Huang1,2,3, Pedro Saavedra1,2
1VIB Center for Inflammation Research, Zwijnaarde, Belgium.
Background And Objective:
Familial Mediterranean fever (FMF) is the most common monogenic autoinflammatory disease (AID) worldwide. The disease is caused by mutations in the MEFV gene encoding the inflammasome sensor Pyrin. Clinical diagnosis of FMF is complicated by overlap in symptoms with other diseases, and interpretation of genetic testing is confounded by the lack of a clear genotype-phenotype association for most of the 340 reported MEFV variants. In this study, the authors designed a functional assay and evaluated its potential in supporting FMF diagnosis.
Methods:
Peripheral blood mononuclear cells (PBMCs) were obtained from patients with Pyrin-associated autoinflammation with an FMF phenotype (n=43) or with autoinflammatory features not compatible with FMF (n=8), 10 asymptomatic carriers and 48 healthy donors. Sera were obtained from patients with distinct AIDs (n=10), and whole blood from a subset of patients and controls. The clinical, demographic, molecular genetic factors and other characteristics of the patient population were assessed for their impact on the diagnostic test read-out. Interleukin (IL)-1β and IL-18 levels were measured by Luminex assay.
Results:
The ex vivo colchicine assay may be performed on whole blood or PBMC. The functional assay robustly segregated patients with FMF from healthy controls and patients with related clinical disorders. The diagnostic test distinguished patients with classical FMF mutations (M694V, M694I, M680I, R761H) from patients with other MEFV mutations and variants (K695R, P369S, R202Q, E148Q) that are considered benign or of uncertain clinical significance.
Conclusion:
The ex vivo colchicine assay may support diagnosis of FMF and functional subtyping of Pyrin-associated autoinflammation.
Insights
A new functional assay using colchicine effectively distinguishes Familial Mediterranean Fever (FMF) patients from healthy individuals and those with similar autoinflammatory conditions. This test aids in diagnosing FMF and classifying Pyrin-associated autoinflammation based on MEFV gene variants.
Area of Science:
- Genetics and Molecular Biology
- Immunology
- Clinical Diagnostics
Background:
- Familial Mediterranean Fever (FMF) is a prevalent monogenic autoinflammatory disease (AID) caused by MEFV gene mutations affecting Pyrin inflammasome sensor.
- Clinical diagnosis of FMF is challenging due to symptom overlap with other conditions and ambiguous genotype-phenotype correlations for numerous MEFV variants.
- Accurate diagnosis is crucial for effective management of FMF and related autoinflammatory disorders.
Purpose of the Study:
- To develop and evaluate a functional assay for supporting the diagnosis of FMF.
- To assess the assay's ability to differentiate FMF patients from healthy controls and individuals with other autoinflammatory conditions.
- To investigate the assay's utility in functional subtyping of Pyrin-associated autoinflammation.
Main Methods:
- Peripheral blood mononuclear cells (PBMCs) and whole blood were collected from FMF patients, individuals with other autoinflammatory features, asymptomatic carriers, and healthy donors.
- An ex vivo colchicine assay was performed on whole blood or PBMCs.
- Interleukin (IL)-1β and IL-18 levels were measured, and clinical/genetic factors were analyzed for their impact on assay results.
Main Results:
- The ex vivo colchicine assay demonstrated robust segregation between FMF patients and healthy controls, as well as patients with related disorders.
- The functional assay successfully distinguished patients with classical FMF-associated MEFV mutations from those with variants of uncertain significance.
- The assay showed potential for differentiating between various Pyrin-associated autoinflammatory phenotypes.
Conclusions:
- The ex vivo colchicine assay is a promising functional tool to aid in the diagnosis of Familial Mediterranean Fever.
- This assay can support the functional subtyping of Pyrin-associated autoinflammation, offering a more nuanced understanding of disease mechanisms.
- The findings suggest a valuable role for functional testing in the diagnostic workup of FMF and related autoinflammatory diseases.
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