Functional evaluation of the pathological significance of MEFV variants using induced pluripotent stem cell–derived macrophages

• Published in: Journal of allergy and clinical immunology Vol. 144; no. 5; pp. 1438 - 1441.e12
• Main Authors: Shiba, Takeshi, Tanaka, Takayuki, Ida, Hiroaki, Watanabe, Misa, Nakaseko, Haruna, Osawa, Mitsujiro, Shibata, Hirofumi, Izawa, Kazushi, Yasumi, Takahiro, Kawasaki, Yuri, Saito, Megumu K., Takita, Junko, Heike, Toshio, Nishikomori, Ryuta
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.11.2019; Elsevier Limited
• Subjects: Apoptosis; Cell death; Cytokines; Fever; Genetic diversity; Genetic variance; Genotype & phenotype; Heredity; IL-1β; Inflammasomes; Internet; Lipopolysaccharides; Macrophages; Monocytes; Mutation; Patients; Peripheral blood; Phenotypes; Pluripotency; Polymorphism; Pyrin protein; Repositories; Stem cells; Stimulation
• ISSN: 0091-6749; 1097-6825
• DOI: 10.1016/j.jaci.2019.07.039

Next, the IL-1β secretion from macrophages derived from monocytes in vitro (peripheral blood–derived macrophages [PB-MPs]) was evaluated. Because TcdA stimulation alone did not induce IL-1β secretion from PB-MPs (data not shown), PB-MPs were primed with LPS before TcdA stimulation. [...]by examining both cell types from the same patients, we revealed the overall picture of the cytokine responses of monocytes and macrophages from patients with FMF. Induced pluripotent stem cell (iPSC) technology provides the opportunity to analyze the effect of genetic variants free from the influence of medication or differences in genetic background. [...]we evaluated whether macrophages derived from patients' iPSCs (iPSC-derived macrophages [iPS-MPs]) recapitulated the phenotype of PB-MPs from patients with FMF. iPSC lines from 3 patients with FMF with the M694I and E148Q MEFV variants were established (patients 6-8; see Fig E2 and Table E1 in this article's Online Repository at www.jacionline.org) and differentiated into iPS-MPs (see Fig E3 in this article's Online Repository at www.jacionline.org). iPS-MPs with the M694I mutation recapitulated the enhanced pyrin inflammasome activation of PB-MPs, leading to increased IL-1β secretion (Fig 2, A), ASC speck formation (Fig 2, B), and cell death, which was dependent on MEFV expression (see Fig E4 in this article's Online Repository at www.jacionline.org). [...]we applied our newly established method to 2 additional MEFV variants, T577N and N679H, which were identified in 2 families in which autoinflammatory disease with dominant inheritance was suspected.