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Immunodeficiency associated with a novel functionally defective variant of SLC19A1 benefits from folinic acid treatment

Abstract

Insufficient dietary folate intake, hereditary malabsorption, or defects in folate transport may lead to combined immunodeficiency (CID). Although loss of function mutations in the major intestinal folate transporter PCFT/SLC46A1 was shown to be associated with CID, the evidence for pathogenic variants of RFC/SLC19A1 resulting in immunodeficiency was lacking. We report two cousins carrying a homozygous pathogenic variant c.1042 G > A, resulting in p.G348R substitution who showed symptoms of immunodeficiency associated with defects of folate transport. SLC19A1 expression by peripheral blood mononuclear cells (PBMC) was quantified by real-time qPCR and immunostaining. T cell proliferation, methotrexate resistance, NK cell cytotoxicity, Treg cells and cytokine production by T cells were examined by flow cytometric assays. Patients were treated with and benefited from folinic acid. Studies revealed normal NK cell cytotoxicity, Treg cell counts, and naive-memory T cell percentages. Although SLC19A1 mRNA and protein expression were unaltered, remarkably, mitogen induced-T cell proliferation was significantly reduced at suboptimal folic acid and supraoptimal folinic acid concentrations. In addition, patients’ PBMCs were resistant to methotrexate-induced apoptosis supporting a functionally defective SLC19A1. This study presents the second pathogenic SLC19A1 variant in the literature, providing the first experimental evidence that functionally defective variants of SLC19A1 may present with symptoms of immunodeficiency.

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Fig. 1: G348R substitution does not alter SLC19A1 protein expression.
Fig. 2: G348R substitution in SLC19A1 reduces T cell proliferation in nonoptimal folate concentrations.
Fig. 3: G348R substitution in SLC19A1 leads to methotrexate resistance.
Fig. 4: Normal NK cell cytotoxicity, normal Treg cell numbers, but increased GM-CSF and IL-17A producing T cells in both patients.

Data availability

All the raw data are available upon reasonable request from the corresponding authors.

References

  1. Shulpekova Y, Nechaev V, Kardasheva S, Sedova A, Kurbatova A, Bueverova E, et al. The concept of folic acid in health and disease. Molecules. 2021;26:3731.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Scaglione F, Panzavolta G. Folate, folic acid and 5-methyltetrahydrofolate are not the same thing. Xenobiotica. 2014;44:480–8.

    Article  CAS  PubMed  Google Scholar 

  3. Field MS, Kamynina E, Chon J, Stover PJ. Nuclear Folate Metabolism. Annu Rev Nutr. 2018;38:219–43.

    Article  CAS  PubMed  Google Scholar 

  4. Fernley RT, Iliades P, Macreadie I. A rapid assay for dihydropteroate synthase activity suitable for identification of inhibitors. Anal Biochem. 2007;360:227–34.

    Article  CAS  PubMed  Google Scholar 

  5. Fox JT, Stover PJ. Folate-mediated one-carbon metabolism. Vitam Horm. 2008;79:1–44.

    Article  CAS  PubMed  Google Scholar 

  6. Crider KS, Yang TP, Berry RJ, Bailey LB. Folate and DNA methylation: a review of molecular mechanisms and the evidence for folate’s role. Adv Nutr. 2012;3:21–38.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Courtemanche C, Elson-Schwab I, Mashiyama ST, Kerry N, Ames BN. Folate deficiency inhibits the proliferation of primary human CD8 + T lymphocytes in vitro. J Immunol. 2004;173:3186–92.

    Article  CAS  PubMed  Google Scholar 

  8. Mikkelsen K, Apostolopoulos V Vitamin B12, folic acid and the immune system. In: Mahmoudi M, Rezaei N, editors. Nutrition and Immunity. Switzerland: Springer; 2019. pp 103–14.

  9. Koury MJ, Ponka P. New insights into erythropoiesis: the roles of folate, vitamin B12, and iron. Annu Rev Nutr. 2004;24:105–31.

    Article  CAS  PubMed  Google Scholar 

  10. Hou Z, Matherly LH. Biology of the major facilitative folate transporters SLC19A1 and SLC46A1. Curr Top Membr. 2014;73:175–204.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Alam C, Kondo M, O’Connor DL, Bendayan R. Clinical implications of folate transport in the central nervous system. Trends Pharm Sci. 2020;41:349–61.

    Article  CAS  PubMed  Google Scholar 

  12. Matherly LH, Wilson MR, Hou Z. The major facilitative folate transporters solute carrier 19A1 and solute carrier 46A1: Biology and role in antifolate chemotherapy of cancer. Drug Metab Dispos. 2014;42:632–49.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Desmoulin SK, Hou Z, Gangjee A, Matherly LH. The human proton-coupled folate transporter: Biology and therapeutic applications to cancer. Cancer Biol Ther. 2012;13:1355–73.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Antony AC. Folate receptors. Annu Rev Nutr. 1996;16:501–21.

    Article  CAS  PubMed  Google Scholar 

  15. Qiu A, Jansen M, Sakaris A, Min SH, Chattopadhyay S, Tsai E, et al. Identification of an intestinal folate transporter and the molecular basis for hereditary folate malabsorption. Cell. 2006;127:917–28.

    Article  CAS  PubMed  Google Scholar 

  16. Borzutzky A, Crompton B, Bergmann AK, Giliani S, Baxi S, Martin M, et al. Reversible severe combined immunodeficiency phenotype secondary to a mutation of the proton-coupled folate transporter. Clin Immunol. 2009;133:287–94.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Kishimoto K, Kobayashi R, Sano H, Suzuki D, Maruoka H, Yasuda K, et al. Impact of folate therapy on combined immunodeficiency secondary to hereditary folate malabsorption. Clin Immunol [Internet]. 2014;153:17–22.

    Article  CAS  PubMed  Google Scholar 

  18. Balashova OA, Visina O, Borodinsky LN. Folate action in nervous system development and disease. Dev Neurobiol. 2018;78:391–402.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Grapp M, Wrede A, Schweizer M, Hüwel S, Galla HJ, Snaidero N, et al. Choroid plexus transcytosis and exosome shuttling deliver folate into brain parenchyma. Nat Commun. 2013;4:2123.

    Article  PubMed  Google Scholar 

  20. Birn H, Spiegelstein O, Christensen EI, Finnell RH. Renal tubular reabsorption of folate mediated by folate binding protein 1. J Am Soc Nephrol. 2005;16:608–15.

    Article  CAS  PubMed  Google Scholar 

  21. Grapp M, Just IA, Linnankivi T, Wolf P, Lücke T, Häusler M, et al. Molecular characterization of folate receptor 1 mutations delineates cerebral folate transport deficiency. Brain. 2012;135:2022–31.

  22. Tang LS, Santillano DR, Wlodarczyk BJ, Miranda RC, Finnell RH. Role of Folbp1 in the regional regulation of apoptosis and cell proliferation in the developing neural tLückeube and craniofacies. Am J Med Genet C Semin Med Genet. 2005;135C:48–58.

    Article  PubMed  Google Scholar 

  23. Spiegelstein O, Mitchell LE, Merriweather MY, Wicker NJ, Zhang Q, Lammer EJ, et al. Embryonic development of folate binding protein-1 (Folbp1) knockout mice: Effects of the chemical form, dose, and timing of maternal folate supplementation. Dev Dyn. 2004;231:221–31.

    Article  CAS  PubMed  Google Scholar 

  24. Zhu H, Cabrera RM, Wlodarczyk BJ, Bozinov D, Wang D, Schwartz RJ, et al. Differentially expressed genes in embryonic cardiac tissues of mice lacking Folr1 gene activity. BMC Dev Biol. 2007;7:128.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Yang-Feng TL, Ma YY, Liang R, Prasad PD, Leibach FH, Ganapathy V. Assignment of the human folate transporter gene to chromosome 21q22.3 by somatic cell hybrid analysis and in situ hybridization. Biochem Biophys Res Commun. 1995;210:874–9.

    Article  CAS  PubMed  Google Scholar 

  26. Kotnik BF, Jazbec J, Grabar PB, Rodriguez-Antona C, Dolzan V. Association between SLC19A1 gene polymorphism and high dose methotrexate toxicity in childhood acute lymphoblastic leukaemia and non hodgkin malignant lymphoma: introducing a haplotype based approach. Radio Oncol. 1751;20:455–62.

    Google Scholar 

  27. Coppedè F, Stoccoro A, Tannorella P, Gallo R, Nicolì V, Migliore L. Association of polymorphisms in genes involved in one-carbon metabolism with MTHFR methylation levels. Int J Mol Sci. 2019;20:3754.

    Article  PubMed  PubMed Central  Google Scholar 

  28. Gelineau-van Waes J, Heller S, Bauer LK, Wilberding J, Maddox JR, Aleman F, et al. Embryonic development in the reduced folate carrier knockout mouse is modulated by maternal folate supplementation. Birth Defects Res A Clin Mol Teratol. 2008;82:494–507.

    Article  PubMed  Google Scholar 

  29. Zhao R, Russell RG, Wang Y, Liu L, Gao F, Kneitz B, et al. Rescue of embryonic lethality in reduced folate carrier-deficient mice by maternal folic acid supplementation reveals early neonatal failure of hematopoietic organs. J Biol Chem. 2001;276:10224–8.

    Article  CAS  PubMed  Google Scholar 

  30. Svaton M, Skvarova Kramarzova K, Kanderova V, Mancikova A, Smisek P, Jesina P, et al. A homozygous deletion in the SLC19A1 gene as a cause of folate-dependent recurrent megaloblastic anemia. Blood 2020;135:2427–31.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Tangye SG, Al-Herz W, Bousfiha A, Cunningham-Rundles C, Franco JL, Holland SM, et al. Human Inborn Errors of Immunity: 2022 Update on the Classification from the International Union of Immunological Societies Expert Committee. J Clin Immunol. 2022:1–35.

  32. Ferguson PL, Flintoff WF. Topological and functional analysis of the human reduced folate carrier by hemagglutinin epitope insertion. J Biol Chem. 1999;274:16269–78.

    Article  CAS  PubMed  Google Scholar 

  33. Alnabbat KI, Fardous AM, Cabelof DC, Heydari AR. Excessive folic acid mimics folate deficiency in human lymphocytes. Curr Issues Mol Biol. 2022;44:1452–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Sharma R, Ali T, Kaur J. Folic acid depletion as well as oversupplementation helps in the progression of hepatocarcinogenesis in HepG2 cells. Sci Rep. 2022;12:16617.

    Article  PubMed  PubMed Central  Google Scholar 

  35. Zhang M, Bracaglia C, Prencipe G, Bemrich-Stolz CJ, Beukelman T, Dimmitt RA, et al. A heterozygous RAB27A mutation associated with delayed cytolytic granule polarization and hemophagocytic lymphohistiocytosis. J Immunol. 2016;196:2492–503.

    Article  CAS  PubMed  Google Scholar 

  36. Kinoshita M, Kayama H, Kusu T, Yamaguchi T, Kunisawa J, Kiyono H, et al. Dietary folic acid promotes survival of Foxp3+ regulatory T cells in the colon. J Immunol. 2012;189:2869–78.

    Article  CAS  PubMed  Google Scholar 

  37. Luteijn RD, Zaver SA, Gowen BG, Wyman SK, Garelis NE, Onia L, et al. SLC19A1 transports immunoreactive cyclic dinucleotides. Nature. 2019;573:434–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Ritchie C, Cordova AF, Hess GT, Bassik MC, Li L. SLC19A1 is an importer of the immunotransmitter cGAMP. Mol Cell. 2019;75:372-81.e5.

    Article  Google Scholar 

  39. Ifergan I, Jansen G, Assaraf YG. The reduced folate carrier (RFC) is cytotoxic to cells under conditions of severe folate deprivation. RFC as a double edged sword in folate homeostasis. J Biol Chem. 2008;283:20687–95.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Yilmaz E, Özcan A, Gök V, Karakükçü M, Ünal E. The effect of methylenetetrahydrofolate reductase polymorphisms on the methotrexate toxicity in children with acute lymphoblastic leukemia: Methylenetetrahydrofolate reductase & methotrexate toxicity. J Transl Pract Med. 2022;1:9–13.

  41. Wu CH, Huang TC, Lin BF. Folate deficiency affects dendritic cell function and subsequent T helper cell differentiation. J Nutr Biochem. 2017;41:65–72.

    Article  CAS  PubMed  Google Scholar 

  42. Besci Ö, Baser D, Öğülür İ, Berberoğlu AC, Kıykım A, Besci T, et al. Reference values for T and B lymphocyte subpopulations in Turkish children and adults. Turk J Med Sci. 2021;51:1814–24.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This study was financially supported by Erciyes University BAP grant (TCD-2021-10863) to EÜ, and Turkish Academy of Science GEBIP and Science Academy BAGEP awards to AE.

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Conceptualization was performed by AE, EÜ. Methodology was performed by ŞE, YH, SB, KEB, CA, AE, MFC. Software was performed by AB. Investigation was performed by VG, ŞE, AÖ, EY, HP, TP, AE, EÜ. Validation was done by AB, MK, AE. Supervision was performed HC, AE, EÜ. Resources—Writing—original draft were written by VG, ŞE, YH, AE. Writing—review & editing were performed by VG, YH, SB, AÖ, EY, CA, MK, HC, HP, TP, AE, EÜ. Funding acquisition by AE, EÜ. All authors have read and approved the manuscript.

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Correspondence to Ahmet Eken or Ekrem Ünal.

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All experimental procedures were approved by Erciyes University institutional review board (#2021/17) and conducted according to regulations. Parents of the patients signed the informed consents for the immunological study and provided consent for the publication of data. Controls were selected randomly from amongst the age and sex-matched healthy donors.

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Gök, V., Erdem, Ş., Haliloğlu, Y. et al. Immunodeficiency associated with a novel functionally defective variant of SLC19A1 benefits from folinic acid treatment. Genes Immun 24, 12–20 (2023). https://doi.org/10.1038/s41435-022-00191-7

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  • DOI: https://doi.org/10.1038/s41435-022-00191-7

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