title: Three-dimensional cellular microarray for high-throughput toxicology assays.
abstract: We have developed a miniaturized 3D cell-culture array (the Data Analysis Toxicology Assay Chip or DataChip) for high-throughput toxicity screening of drug candidates and their cytochrome P450-generated metabolites. The DataChip consists of human cells encapsulated in collagen or alginate gels (as small as 20 nl) arrayed on a functionalized glass slide for spatially addressable screening against multiple compounds. A single DataChip containing 1,080 individual cell cultures, used in conjunction with the complementary human P450-containing microarray (the Metabolizing Enzyme Toxicology Assay Chip or MetaChip), simultaneously provided IC(50) values for nine compounds and their metabolites from CYP1A2, CYP2D6, and CYP3A4 and a mixture of the three P450s designed to emulate the human liver. Similar responses were obtained with the DataChip and conventional 96-well plate assays, demonstrating that the near 2,000-fold miniaturization does not influence the cytotoxicity response. The DataChip may therefore enable toxicity analyses of drug candidates and their metabolites at throughputs compatible with the availability of compounds at early-stage drug discovery.
authors.affiliation: Solidus Biosciences, Inc., Troy, NY 12180, USA.
authors.forename: Moo-Yeal
authors.initials: MY
authors.lastname: Lee
authors.forename: R Anand
authors.initials: RA
authors.lastname: Kumar
authors.forename: Sumitra M
authors.initials: SM
authors.lastname: Sukumaran
authors.forename: Michael G
authors.initials: MG
authors.lastname: Hogg
authors.forename: Douglas S
authors.initials: DS
authors.lastname: Clark
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Dordick
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002626:Chemistry, Pharmaceutical / Q000379:methods; D003094:Collagen / Q000378:metabolism; D019388:Cytochrome P-450 CYP1A2 / Q000737:chemistry; D019389:Cytochrome P-450 CYP2D6 / Q000737:chemistry; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry; D015195:Drug Design; D004867:Equipment Design; D006801:Humans; D020128:Inhibitory Concentration 50; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D010597:Pharmacogenetics / Q000295:instrumentation / Q000379:methods*; D051381:Rats; D013678:Technology, Pharmaceutical / Q000639:trends; D014116:Toxicology / Q000379:methods*
chemical_list: D003094:Collagen; D003577:Cytochrome P-450 Enzyme System; D019388:Cytochrome P-450 CYP1A2; D019389:Cytochrome P-450 CYP2D6; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human