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.
authors.affiliation: School of Medicine & Dentistry, University of Aberdeen, Aberdeen, Scotland, United Kingdom.
authors.forename: Karolin
authors.initials: K
authors.lastname: Hijazi
authors.affiliation: Laboratory of Molecular Microbiology and Biotechnology, Department of Medical Biotechnologies, University of Siena, Siena, Italy.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Cuppone
authors.affiliation: School of Medicine & Dentistry, University of Aberdeen, Aberdeen, Scotland, United Kingdom.
authors.forename: Kieron
authors.initials: K
authors.lastname: Smith
authors.affiliation: MICROBIOTEC srl, Siena, Italy.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Stincarelli
authors.affiliation: Mucosal Infection & Immunity Group, Section of Infectious Diseases, Imperial College, London, United Kingdom.
authors.forename: Julia
authors.initials: J
authors.lastname: Ekeruche-Makinde
authors.affiliation: Laboratory of Molecular Microbiology and Biotechnology, Department of Medical Biotechnologies, University of Siena, Siena, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: De Falco
authors.affiliation: School of Medicine & Dentistry, University of Aberdeen, Aberdeen, Scotland, United Kingdom.
authors.forename: Georgina L
authors.initials: GL
authors.lastname: Hold
authors.affiliation: Mucosal Infection & Immunity Group, Section of Infectious Diseases, Imperial College, London, United Kingdom.
authors.forename: Robin
authors.initials: R
authors.lastname: Shattock
authors.affiliation: Dental Institute, King's College London, London, United Kingdom.
authors.forename: Charles G
authors.initials: CG
authors.lastname: Kelly
authors.affiliation: Laboratory of Molecular Microbiology and Biotechnology, Department of Medical Biotechnologies, University of Siena, Siena, Italy; MICROBIOTEC srl, Siena, Italy.
authors.forename: Gianni
authors.initials: G
authors.lastname: Pozzi
authors.affiliation: Laboratory of Molecular Microbiology and Biotechnology, Department of Medical Biotechnologies, University of Siena, Siena, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Iannelli
mesh_terms: D000328:Adult; D019380:Anti-HIV Agents / Q000494:pharmacology*; D001692:Biological Transport; D002460:Cell Line; D045744:Cell Line, Tumor; D002584:Cervix Uteri / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D000069454:Darunavir / Q000494:pharmacology; D004351:Drug Resistance; D004357:Drug Synergism; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D026901:Membrane Transport Proteins / Q000096:biosynthesis* / Q000235:genetics; D008875:Middle Aged; D009924:Organ Culture Techniques; D011743:Pyrimidines / Q000494:pharmacology; D000068698:Tenofovir / Q000494:pharmacology; D014621:Vagina / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
chemical_list: D019380:Anti-HIV Agents; D026901:Membrane Transport Proteins; D011743:Pyrimidines; D000068698:Tenofovir; C481671:Dapivirine; D000069454:Darunavir