Abstract
Parkinson’s disease is a multifactorial disease; both genetic predisposition (5% of all cases), environmental factors, and age-related changes in the brain and other body systems contribute to its etiology. For the diagnosis and study of the pathology of the disease development, it is important to search for new polymorphisms associated with hereditary forms of the disease. The clinical exome of a 55-year-old patient with Parkinson’s disease was analyzed and a single nucleotide polymorphism in the GLUD2 gene (c.1492T>G) was identified. This genetic variant is pathogenic according to the ClinVar database, but the mechanism of pathogenesis is still poorly understood. In addition, there are currently no relevant models based on human cells, which is of great interest. We generated induced pluripotent stem cells (iPSCs) from patient peripheral blood mononuclear cells using nonintegrating episomal vectors expressing OCT4, KLF4, L-MYC, SOX2, LIN28, and p53 shRNA. The obtained iPSC lines (ICGi044-B and ICGi044-C) demonstrate typical ESC-like morphology, normal karyotype (46,XY), express pluripotency markers (OCT4, SOX2, NANOG, SSEA4, TRA‑1-60), and are able to give derivatives of three germ layers. The iPSC lines ICGi044-B and ICGi044-C, as well as their neural derivatives, represent a unique in vitro cell model for studying the pathogenetic mechanisms of the development of Parkinson’s disease associated with the c.1492T>G mutation in the GLUD2 gene.
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ACKNOWLEDGMENTS
Immunofluorescent visualization was performed using the resources of the Center for Collective Use of Microscopic Analysis of Biological Objects of the Institute of Cytology and Genetics, Siberian Branch, Russian Academy of Sciences (https://ckp.icgen.ru/ckpmabo/), supported by the Budgetary Project of the Institute of Cytology and Genetics, Siberian Branch, Russian Academy of Sciences, FWNR-2022-0015. PBMCs were provided by the Federal State Budgetary Institution Federal Center for Neurosurgery of the Ministry of Health of the Russian Federation. Clinical exome sequencing was performed at the Novosibirsk National Research State University.
Funding
The study was financially supported by the Russian Science Foundation, project no. 19-75-20063.
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E.V. Grigor’eva, S.V. Pavlova, A.A. Malakhova, and S.M. Zakian developed the design of the experiment, analyzed the obtained data and participated in writing the article. D.A. Sorogina and E.V. Grigor’eva reprogrammed the patient’s mononuclear cells. All cultural and molecular genetic work was carried out by D.A. Sorogina, namely, obtaining individual iPSC clones and their detailed characterization. S.P. Medvedev performed Sanger sequencing to confirm the presence of polymorphisms in the obtained iPSCs. Analysis of exome sequencing data was performed by Y.V. Vyatkin. The medical support of the patient and the provision of mononuclear cells was carried out by E.A. Khabarova and J.A. Rzaev.
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Statement of compliance with standards of research involving humans as subjects. The study was approved by the ethical commission of the Federal State Budgetary Institution Federal Center for Neurosurgery of the Ministry of Health of the Russian Federation, Novosibirsk, Protocol no. 1 dated March 14, 2017. The patient was provided with all information about this study and signed the informed consent and information sheet with his own hand.
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Passport of a line of pluripotent stem cells
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Sorogina, D.A., Grigor’eva, E.V., Malakhova, A.A. et al. Creation of Induced Pluripotent Stem Cells ICGi044-B and ICGi044-C Using Reprogramming of Peripheral Blood Mononuclear Cells of a Patient with Parkinson’s Disease Associated with c.1492T>G Mutation in the GLUD2 Gene. Russ J Dev Biol 54, 104–111 (2023). https://doi.org/10.1134/S1062360423010125
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DOI: https://doi.org/10.1134/S1062360423010125