title: Direct assessment of drug penetration into tissue using a novel application of three-dimensional cell culture.
abstract: The failure of many anticancer drugs to control growth of solid cancers may stem in part from inadequate delivery to tumor regions distant from vasculature. Although the identification of new anticancer drug targets has led to the development of many new drug candidates, there is a lack of methodology for identifying drugs that adequately penetrate tumor tissue. We have developed a novel multilayered cell culture-based assay, which detects the penetration of anticancer drugs based on their effect within tissue. Drug exposures are made over 1 hour to one side of a disk of tissue approximately 150-microm thick, with the other side temporarily closed off, and penetration is then assessed 1-3 days later via bromodeoxyuridine-based detection of S-phase cells. Using this assay, the tissue distribution of a selection of anthracycline analogues was assessed. At clinically relevant exposures, none of the agents were able to affect cells on the far side of the culture at levels approaching that seen on the near (exposed) side. Doxorubicin and epirubicin exhibited approximately 10-fold decreases in the drug exposure seen by the cells on the far side relative to those on the near side of the cultures, whereas for daunorubicin and mitoxantrone, approximately 30-fold and >30-fold decreases were observed respectively. Results were consistent with the observed gradients in drug-derived fluorescence of doxorubicin, epirubicin, and daunorubicin. This model could be applied as a simple anticancer drug development screen to discover drugs that exhibit desirable penetration properties.
authors.affiliation: Department of Medical Biophysics, British Columbia Cancer Research Centre, Vancouver, British Columbia, Canada.
authors.forename: Alastair H
authors.initials: AH
authors.lastname: Kyle
authors.forename: Lynsey A
authors.initials: LA
authors.lastname: Huxham
authors.forename: Aaron S J
authors.initials: AS
authors.lastname: Chiam
authors.forename: David H
authors.initials: DH
authors.lastname: Sim
authors.forename: Andrew I
authors.initials: AI
authors.lastname: Minchinton
mesh_terms: D018943:Anthracyclines / Q000493:pharmacokinetics*; D000903:Antibiotics, Antineoplastic / Q000493:pharmacokinetics*; D015179:Colorectal Neoplasms / Q000378:metabolism* / Q000473:pathology*; D003630:Daunorubicin / Q000493:pharmacokinetics; D004317:Doxorubicin / Q000493:pharmacokinetics; D015251:Epirubicin / Q000493:pharmacokinetics; D045325:HCT116 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008942:Mitoxantrone / Q000493:pharmacokinetics; D013050:Spectrometry, Fluorescence; D014407:Tumor Cells, Cultured
chemical_list: D018943:Anthracyclines; D000903:Antibiotics, Antineoplastic; D015251:Epirubicin; D004317:Doxorubicin; D008942:Mitoxantrone; D003630:Daunorubicin