Ofelia M Noceti1, Jean-Baptiste Woillard2, Ahmed Boumediene3, Patricia Esperón4, Jean-Luc Taupin5, Solange Gerona6, Marcelo Valverde6, Cristina Touriño7, Pierre Marquet8. 1. INSERM U850 and Liver Diseases Department, National Center for Liver Transplantation, Hospital Central de las Fuerzas Armadas, Montevideo, Uruguay; Molecular Biology Unit, Clinical Biochemistry Department, School of Chemistry, and. 2. INSERM U850 and. 3. CNRS UMR 7276, Université Limoges, Limoges, France; 4. Molecular Biology Unit, Clinical Biochemistry Department, School of Chemistry, and. 5. CNRS UMR 5164, Université Bordeaux Segalen, Bordeaux, France. 6. Liver Diseases Department, National Center for Liver Transplantation, Hospital Central de las Fuerzas Armadas, Montevideo, Uruguay; 7. Department of Fundamental Medicine, School of Medicine, Universidad de la República, Montevideo, Uruguay; 8. INSERM U850 and pierre.marquet@unilim.fr.
Abstract
BACKGROUND: Although therapeutic drug monitoring has improved the clinical use of immunosuppressive drugs, there is still interpatient variability in efficacy and toxicity that pharmacodynamic monitoring may help to reduce. To select the best biomarkers of tacrolimus pharmacodynamics, we explored the strength and variability of signal transduction and the influence of polymorphisms along the calcineurin pathway. METHODS: Peripheral blood mononuclear cells from 35 healthy volunteers were incubated with tacrolimus (0.1-50 ng/mL) and stimulated ex vivo. Inhibition of NFAT1 (nuclear factor of activated T cells 1) translocation to the nucleus and intracellular expression of interleukin-2 in CD4(+) and CD8(+) T cells and the surface activation marker CD25 in CD3(+) cells were measured by flow cytometry. We sequenced the promoter regions of immunophilins and calcineurin subunits and characterized selected single nucleotide polymorphisms in the genes of the calcineurin pathway with allelic discrimination assays. RESULTS: All responses closely fitted an I/Imax sigmoid model. Large interindividual variability (n = 30) in I0 and IC50 was found for all biomarkers. Moreover, strong and statistically significant associations were found between tacrolimus pharmacodynamic parameters and polymorphisms in the genes coding cyclophilin A, the calcineurin catalytic subunit α isoenzyme, and CD25. CONCLUSIONS: This study demonstrates the consistency and large interindividual variability of signal transduction along the calcineurin pathway, as well as the strong influence of pharmacogenetic polymorphisms in the calcineurin cascade on both the physiological activity of this route and tacrolimus pharmacodynamics.
BACKGROUND: Although therapeutic drug monitoring has improved the clinical use of immunosuppressive drugs, there is still interpatient variability in efficacy and toxicity that pharmacodynamic monitoring may help to reduce. To select the best biomarkers of tacrolimus pharmacodynamics, we explored the strength and variability of signal transduction and the influence of polymorphisms along the calcineurin pathway. METHODS: Peripheral blood mononuclear cells from 35 healthy volunteers were incubated with tacrolimus (0.1-50 ng/mL) and stimulated ex vivo. Inhibition of NFAT1 (nuclear factor of activated T cells 1) translocation to the nucleus and intracellular expression of interleukin-2 in CD4(+) and CD8(+) T cells and the surface activation marker CD25 in CD3(+) cells were measured by flow cytometry. We sequenced the promoter regions of immunophilins and calcineurin subunits and characterized selected single nucleotide polymorphisms in the genes of the calcineurin pathway with allelic discrimination assays. RESULTS: All responses closely fitted an I/Imax sigmoid model. Large interindividual variability (n = 30) in I0 and IC50 was found for all biomarkers. Moreover, strong and statistically significant associations were found between tacrolimus pharmacodynamic parameters and polymorphisms in the genes coding cyclophilin A, the calcineurin catalytic subunit α isoenzyme, and CD25. CONCLUSIONS: This study demonstrates the consistency and large interindividual variability of signal transduction along the calcineurin pathway, as well as the strong influence of pharmacogenetic polymorphisms in the calcineurin cascade on both the physiological activity of this route and tacrolimus pharmacodynamics.
Authors: Dirk Jan A R Moes; Rogier R Press; Oliver Ackaert; Bart A Ploeger; Frederike J Bemelman; Cheikh Diack; Judith A M Wessels; Tahar van der Straaten; Meindert Danhof; Jan-Stephan F Sanders; Jaap J Homan van der Heide; Henk Jan Guchelaar; Johan W de Fijter Journal: Br J Clin Pharmacol Date: 2016-05-10 Impact factor: 4.335
Authors: Patricia C Salgado; Fabiana Dv Genvigir; Claudia R Felipe; Helio Tedesco-Silva; Jose O Medina-Pestana; Sonia Q Doi; Mario H Hirata; Rosario Dc Hirata Journal: Pharmgenomics Pers Med Date: 2017-03-31
Authors: Aliede E In 't Veld; Hendrika W Grievink; Mahdi Saghari; Frederik E Stuurman; Marieke L de Kam; Aiko P J de Vries; Brenda C M de Winter; Jacobus Burggraaf; Adam F Cohen; Matthijs Moerland Journal: Int J Mol Sci Date: 2019-09-23 Impact factor: 5.923