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Human Peripheral Blood Macrophages As a Model for Studying Glucocerebrosidase Dysfunction

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Abstract—

Decreased activity of glucocerebrosidase (GCase) as a result of mutations in the GBA gene causes Gaucher’s disease (GD), which belongs to the group of lysosomal storage disorders. The risk of Parkinson’s disease in homo- and heterozygous carriers of GBA mutations is elevated seven- to eightfold. Screening of novel compounds designed to enhance GCase activity requires development of in vitro models based on primary cell cultures obtained from patients carrying GBA mutations. In this work, the efficiency of different methods used to culture peripheral blood macrophages of GD patients and control subjects was compared, and GCase activity and lysosphingolipid concentrations were evaluated using tandem mass spectrometry (HPLC‒MS/MS) in dried cell spots. For the first time, the efficacy of restoring the activity of mutant GCase has been assessed in primary macrophages of GD patients cultured in the presence of pharmacological GCase chaperones isofagomine and ambroxol. Based on these results, a convenient method of in vitro screening of candidate pharmacological agents designed to increase GCase activity can be proposed.

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ACKNOWLEDGMENTS

This work was financially supported by the Russian Science Foundation, project no. 17-75-20159.

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Authors and Affiliations

  1. St. Petersburg Konstantinov Institute of Nuclear Physics, National Research Center Kurchatov Institute, 188300, Gatchina, Leningrad oblast, Russia

    M. A. Nikolaev, A. E. Kopytova, A. K. Emel’yanov, K. A. Senkevich, T. S. Usenko & S. N. Pchelina

  2. Pavlov First St. Petersburg State Medical University, 197022, St. Petersburg, Russia

    M. A. Nikolaev, A. K. Emel’yanov, K. A. Senkevich, T. S. Usenko, M. V. Gorchakova, Yu. P. Koval’chuk, O. A. Berkovich & S. N. Pchelina

  3. Research Center of Medical Genetics, 115522, Moscow, Russia

    G. V. Baidakova & E. Yu. Zakharova

  4. Almazov National Medical Research Center, Ministry of Health of the Russian Federation, 197341, St. Petersburg, Russia

    G. N. Salogub

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  1. M. A. Nikolaev
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  2. A. E. Kopytova
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  3. G. V. Baidakova
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  11. E. Yu. Zakharova
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  12. S. N. Pchelina
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Correspondence to M. A. Nikolaev.

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Conflict of interests. The authors declare that they have no conflict of interest.

Statement of compliance with standards of research involving humans as subjects. The study was approved by the Ethics Committee of Pavlov Medical University; all subjects signed the informed consent form.

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Translated by D. Timchenko

Abbreviations: PD—Parkinson’s disease, GD—Gaucher’s disease, HPLC—high-performance liquid chromatography, MS/MS—tandem mass spectrometry, GCase—glucocerebrosidase, GBA—glucocerebrosidase gene, M-CSF—macrophage colony-stimulating factor, HexSph—hexosylsphingosine.

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Nikolaev, M.A., Kopytova, A.E., Baidakova, G.V. et al. Human Peripheral Blood Macrophages As a Model for Studying Glucocerebrosidase Dysfunction. Cell Tiss. Biol. 13, 100–106 (2019). https://doi.org/10.1134/S1990519X19020081

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  • DOI: https://doi.org/10.1134/S1990519X19020081

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