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

doi:10.1371/journal.pone.0131405

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 language	English
Article number	e0131405
Number of pages	18
Journal	PloS ONE
Volume	10
Issue number	6
Early online date	23 Jun 2015
DOIs	https://doi.org/10.1371/journal.pone.0131405
Publication status	Published - 23 Jun 2015

Bibliographical note

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/158530_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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10.1371/journal.pone.0131405Licence: CC BY

Expression of Genes for Drug Transporters in the Human Female Genital Tract and Modulatory Effect of Antiretroviral Drugs
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by/4.0/
Final published version, 560 KBLicence: CC BY

Karolin Hijazi
- School of Medicine, Medical Sciences & Nutrition, Dental Education - Clinical Chair
Person: Clinical Academic

Cite this

Hijazi, K., Cuppone, A. M., Smith, K., Stincarelli, M. A., Ekeruche-Makinde, J., De Falco, G., Hold, G. L., Shattock, R., Kelly, C. G., 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, 10(6), Article e0131405. https://doi.org/10.1371/journal.pone.0131405

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

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.",

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

note = "Acknowledgments: We wish to thank patients and surgical teams at the University of Siena and St Mary{\textquoteright}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/158530_en.html. Beneficiaries of MOTIF include the University of Aberdeen, the University of Siena, Microbiotec srl, Imperial College, and King{\textquoteright}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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AU - Hijazi, Karolin

AU - Cuppone, Anna M.

AU - Smith, Kieron

AU - Stincarelli, Maria A.

AU - Ekeruche-Makinde, Julia

AU - De Falco, Giulia

AU - Hold, Georgina L.

AU - Shattock, Robin

AU - Kelly, Charles G

AU - Pozzi, Gianni

AU - Iannelli, Francesco

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/158530_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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AB - 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.

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DO - 10.1371/journal.pone.0131405

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M1 - e0131405

ER -

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

Abstract

Bibliographical note

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Karolin Hijazi

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