Generation of iPSCs from a Patient with the M694V Mutation in the MEFV Gene Associated with Familial Mediterranean Fever and Their Differentiation into Macrophages

Elena V Grigor'eva; Lana V Karapetyan; Anastasia A Malakhova; Sergey P Medvedev; Julia M Minina; Varduhi H Hayrapetyan; Valentina S Vardanyan; Suren M Zakian; Arsen Arakelyan; Roksana Zakharyan

doi:10.3390/ijms25116102

Generation of iPSCs from a Patient with the M694V Mutation in the MEFV Gene Associated with Familial Mediterranean Fever and Their Differentiation into Macrophages

Int J Mol Sci. 2024 Jun 1;25(11):6102. doi: 10.3390/ijms25116102.

Authors

Elena V Grigor'eva^{1

2

3}, Lana V Karapetyan⁴, Anastasia A Malakhova^{1

2

3}, Sergey P Medvedev^{1

2

3}, Julia M Minina¹, Varduhi H Hayrapetyan^{4

5}, Valentina S Vardanyan^{6

7}, Suren M Zakian^{1

2

3}, Arsen Arakelyan^{4

5}, Roksana Zakharyan^{4

5}

Affiliations

¹ Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, Russia.
² Meshalkin National Medical Research Center, Ministry of Health of the Russian Federation, 630055 Novosibirsk, Russia.
³ Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, Russia.
⁴ Department of Bioengineering, Bioinformatics, and Molecular Biology, Institute of Biomedicine and Pharmacy, Russian-Armenian (Slavonic) University, Yerevan 0051, Armenia.
⁵ Institute of Molecular Biology NAS RA, Yerevan 0014, Armenia.
⁶ Department of Rheumatology, Yerevan State Medical University after Mkhitar Heratsi (YSMU), Yerevan 0025, Armenia.
⁷ Department of Rheumatology, "Mikaelyan" Institute of Surgery, Yerevan 0052, Armenia.

Abstract

Familial Mediterranean fever (FMF) is a systemic autoinflammatory disorder caused by inherited mutations in the MEFV (Mediterranean FeVer) gene, located on chromosome 16 (16p13.3) and encoding the pyrin protein. Despite the existing data on MEFV mutations, the exact mechanism of their effect on the development of the pathological processes leading to the spontaneous and recurrent autoinflammatory attacks observed in FMF, remains unclear. Induced pluripotent stem cells (iPSCs) are considered an important tool to study the molecular genetic mechanisms of various diseases due to their ability to differentiate into any cell type, including macrophages, which contribute to the development of FMF. In this study, we developed iPSCs from an Armenian patient with FMF carrying the M694V, p.(Met694Val) (c.2080A>G, rs61752717) pathogenic mutation in exon 10 of the MEFV gene. As a result of direct differentiation, macrophages expressing CD14 and CD45 surface markers were obtained. We found that the morphology of macrophages derived from iPSCs of a patient with the MEFV mutation significantly differed from that of macrophages derived from iPSCs of a healthy donor carrying the wild-type MEFV gene.

Keywords: Familial Mediterranean fever; MEFV gene; differentiation; macrophages; patient-specific induced pluripotent stem cells.

MeSH terms

Cell Differentiation* / genetics
Familial Mediterranean Fever* / genetics
Familial Mediterranean Fever* / pathology
Humans
Induced Pluripotent Stem Cells* / metabolism
Macrophages* / metabolism
Male
Mutation*
Pyrin* / genetics

Substances

Pyrin
MEFV protein, human

Grants and funding

The cell reprogramming and characterization was funded by the Ministry of Science and Higher Education of the Russian Federation, Agreement No. 075-15-2021-1063/10. PBMC isolation and molecular-genetic characterization work was supported by the Higher Science and Education Committee of the Ministry of Science, Education, Culture and Sports of the Republic of Armenia, in the frames of the research project N 21SCG-1F010 and research grant provided by the Armenian Engineers and Scientists of America (AESA, PI: Dr. Roksana Zakharyan). The immunofluorescent imaging was performed using resources of the Common Facilities Center of Microscopic Analysis of Biological Objects, ICG SB RAS (https://ckp.icgen.ru/ckpmabo/, accessed on 13 March 2024), supported by the Budget project of the Institute of Cytology and Genetics (FWNR-2022-0015).