Expression of Genes for Drug Transporters in the Human Female Genital Tract and Modulatory Effect of Antiretroviral Drugs

Karolin Hijazi, Anna M. Cuppone, Kieron Smith, Maria A. Stincarelli, Julia Ekeruche-Makinde, Giulia De Falco, Georgina L. Hold, Robin Shattock, Charles G Kelly, Gianni Pozzi, Francesco Iannelli

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Abstract

Anti-retroviral (ARV) –based microbicides are one of the strategies pursued to prevent HIV-1 transmission. Delivery of ARV drugs to subepithelial CD4+ T cells at concentrations for protection is likely determined by drug transporters expressed in the cervicovaginal epithelium. To define the role of drug transporters in mucosal disposition of topically applied ARV-based microbicides, these must be tested in epithelial cell line-based biopharmaceutical assays factoring the effect of relevant drug transporters. We have characterised gene expression of influx and efflux drug transporters in a panel of cervicovaginal cell lines and compared this to expression in cervicovaginal tissue. We also investigated the effect of dapivirine, darunavir and tenofovir, currently at advanced stages of microbicides development, on expression of drug transporters in cell lines. Expression of efflux ABC transporters in cervical tissue was best represented in HeLa, Ect1/E6E7 and End1/E6E7 cell lines. Expression of influx OCT and ENT transporters in ectocervix matched expression in Hela while expression of influx SLCO transporters in vagina was best reflected in VK2/E6E7 cell line. Stimulation with darunavir and dapivirine upregulated MRP transporters, including MRP5 involved in transport of tenofovir. Dapivirine also significantly downregulated tenofovir substrate MRP4 in cervical cell lines. Treatment with darunavir and dapivirine showed no significant effect on expression of BCRP, MRP2 and P-glycoprotein implicated in efflux of different ARV drugs. Darunavir strongly induced expression in most cell lines of CNT3 involved in cell uptake of nucleotide/nucleoside analogue reverse transcriptase inhibitors and SLCO drug transporters involved in cell uptake of protease inhibitors. This study provides insight into the suitability of cervicovaginal cell lines for assessment of ARV drugs in transport kinetics studies. The modulatory effect of darunavir and dapivirine on expression of drug transporters involved in transport of tenofovir points to the possibility of combining these drugs to improve retention of individual drugs at target tissues.
Original languageEnglish
Article numbere0131405
Number of pages18
JournalPloS ONE
Volume10
Issue number6
Early online date23 Jun 2015
DOIs
Publication statusPublished - 23 Jun 2015

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female genitalia
transporters
Tenofovir
Genes
Gene Expression
drugs
gene expression
cell lines
Pharmaceutical Preparations
Cells
Cell Line
Anti-Infective Agents
anti-infective agents
Tissue
antiretroviral agents
uptake mechanisms
biopharmaceuticals
ABC transporters
RNA-directed DNA polymerase
nucleosides

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Hijazi, K., Cuppone, A. M., Smith, K., Stincarelli, M. A., Ekeruche-Makinde, J., De Falco, G., ... Iannelli, F. (2015). Expression of Genes for Drug Transporters in the Human Female Genital Tract and Modulatory Effect of Antiretroviral Drugs. PloS ONE, 10(6), [e0131405]. https://doi.org/10.1371/journal.pone.0131405

Expression of Genes for Drug Transporters in the Human Female Genital Tract and Modulatory Effect of Antiretroviral Drugs. / Hijazi, Karolin; Cuppone, Anna M. ; Smith, Kieron; Stincarelli, Maria A. ; Ekeruche-Makinde, Julia ; De Falco, Giulia; Hold, Georgina L.; Shattock, Robin; Kelly, Charles G; Pozzi, Gianni; Iannelli, Francesco .

In: PloS ONE, Vol. 10, No. 6, e0131405, 23.06.2015.

Research output: Contribution to journalArticle

Hijazi, K, Cuppone, AM, Smith, K, Stincarelli, MA, Ekeruche-Makinde, J, De Falco, G, Hold, GL, Shattock, R, Kelly, CG, Pozzi, G & Iannelli, F 2015, 'Expression of Genes for Drug Transporters in the Human Female Genital Tract and Modulatory Effect of Antiretroviral Drugs', PloS ONE, vol. 10, no. 6, e0131405. https://doi.org/10.1371/journal.pone.0131405
Hijazi, Karolin ; Cuppone, Anna M. ; Smith, Kieron ; Stincarelli, Maria A. ; Ekeruche-Makinde, Julia ; De Falco, Giulia ; Hold, Georgina L. ; Shattock, Robin ; Kelly, Charles G ; Pozzi, Gianni ; Iannelli, Francesco . / Expression of Genes for Drug Transporters in the Human Female Genital Tract and Modulatory Effect of Antiretroviral Drugs. In: PloS ONE. 2015 ; Vol. 10, No. 6.
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N1 - Acknowledgments: We wish to thank patients and surgical teams at the University of Siena and St Mary’s Hospital London for their generous contribution to this study. Darunavir was kindly provided by Janssen R&D Ireland, and tenofovir by Gilead Sciences. Funding: This study was supported by the European Commission grant MOTIF (Microbicide Optimization Through Innovative Formulation for Vaginal and Rectal Delivery), FP7-HEALTH-2012-305316 http://cordis.europa.eu/result/rcn/15853​0_en.html. Beneficiaries of MOTIF include the University of Aberdeen, the University of Siena, Microbiotec srl, Imperial College, and King’s College London. MICROBIOTEC srl provided support in the form of a salary for MAS, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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