NF-kB pathway hyperactivation and its interplay with CD40L on Behçet's Disease: a possible link for the autoinflammatory phenotype

• Grant number: 18/00555-6
• Support Opportunities: Research Grants - Young Investigators Grants
• Start date: November 01, 2018
• End date: April 30, 2024
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Sandro Félix Perazzio
• Grantee: Sandro Félix Perazzio
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated researchers: Luiz Eduardo Coelho Andrade
• Associated scholarship(s): 19/23469-0 - NF-kB pathway hyperactivation and its interplay with CD40L on Behçets Disease: a possible link for the autoinflammatory phenotype, BP.MS; 19/13814-2 - NF-kB pathway hyperactivation and its interplay with CD40L on Behçet's Disease: a possible link for the autoinflammatory phenotype, BP.MS

Abstract

Behçet's Disease (BD) is currently classified as a polygenic autoinflammatory syndrome. Recently point mutations in NFKB1, TNFAIP3 and NEMO were associated with BD-like phenotypes, showing the importance of the NF-kB pathway on BD pathogenesis. Constitutional or acquired trisomy 8, which encodes IKBKB, an important NF-kB activator, is often associated with BD. In addition, sCD40L, which, in turn, has as abnormal plasma concentration in BD, can stimulate oxidative burst via PI3K/NF-kB. However, the influence of the above-mentioned mutations in immune cells and sCD40L-mediated signaling remain unclear. Purpose: 1) analyze the functional behavior of HL-60, THP-1, Daudi, Jurkat and U293F cell lines influenced by NF-kB pathway components hyperexpression and mutations, focusing on sCD40L-induced activation; 2) create a BD-like mouse model by injecting high doses of sCD40L. Methods: 1) in vitro study: lentiviruses labeled with green fluorescent protein carrying NFKB1, NFKB2, NEMO, TNFAIP3 and IKBKB and their respective mutant variants R157X, R853X, D406V and L227X will transduce the cell lines. The hyperexpression of the NF-kB pathways wild type and mutant genes transduced will be confirmed by flow cytometry. Cell lines will be assessed regarding: neutrophil extracellular traps release; oxidative burst; phagocytic; in vitro production of TNFa, IL-1b e IL-6; genetic and protein expression of NF-kB pathway components and target genes; 2) experimental study: CD40L-deficient and C57BL/6J mice will receive weekly intraperitoneal injections of sCD40L for 90 days. Plasma levels of sCD40L will be determined every other week by ELISA. After this period, the animals will be sacrificed for a comprehensive histopathological study and exsanguinated for quantification of serum autoantibodies. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

A.L.S. CUNHA; S.F. PERAZZIO. Effects of immune exhaustion and senescence of innate immunity in autoimmune disorders. Brazilian Journal of Medical and Biological Research, v. 57, 2024-06-17. (18/00555-6, 19/23469-0)