Development of Assays for the Expression of Mutated Pyrin (MEFV)

• Grant number: 25/17298-0
• Support Opportunities: Regular Research Grants
• Start date: January 01, 2026
• End date: December 31, 2026
• Field of knowledge: Biological Sciences - Immunology - Immunogenetics
• Principal Investigator: Alessandra Pontillo
• Grantee: Alessandra Pontillo
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• City of the host institution: São Paulo
• Associated research grant: 23/17878-0 - Evaluation of Inflammasome Activation in Cells of Patients with Autoinflammatory Disease and Its Correlation with Individual Genetics, AP.R

Abstract

Pyrin-associated autoinflammatory diseases, or pyrinopathies, are systemic inflammatory disorders caused by gain-of-function mutations in the MEFV gene, which encodes the pyrin inflammasome receptor. These mutations lead to hyperactivation of the inflammasome and increased production of the inflammatory cytokines IL-1¿ and/or IL-18, along with induction of pyroptosis, resulting in chronic systemic inflammation.The most common and "classic" syndrome is the autosomal recessive form known as Familial Mediterranean Fever (FMF). However, more recently, both dominant and recessive forms with distinct clinical phenotypes-such as PAAND, PAAD, PAANI, and PAAHE-have been described. These findings increase the diagnostic complexity and point to still unknown mechanisms of pyrin inflammasome regulation.Although genetic analysis remains the primary diagnostic tool, functional assays to characterize mutations and novel variants are essential to deepen our understanding of the pyrin inflammasome, assess prognosis, and guide therapeutic decisions.In this project, we propose to study pyrin mutations through overexpression of the mutated gene in a cellular system lacking an inflammasome (HEK-293T) and in a system that possesses an active inflammasome (THP-1). For this, we will use standard plasmid and lentiviral vectors to express both the wild-type (WT) and mutated MEFV gene in HEK-293T and THP-1 cells, respectively. We have selected two mutations: a "classic" one (M694V) associated with FMF, and a more recently reported one (S242R) associated with PAAND (https://infevers.umai-montpellier.fr). The readouts of pyrin activation will be pyrin/ASC oligomerization (speck formation) and IL-1¿ release.The fellowship recipient will have the opportunity to learn and perform molecular biology and cell culture techniques as tools for translational research, as well as to engage in the dynamics of a research group, including literature review, data presentation, and scientific article discussions. (AU)