title: 3-Dimensional culture systems for anti-cancer compound profiling and high-throughput screening reveal increases in EGFR inhibitor-mediated cytotoxicity compared to monolayer culture systems.
abstract: 3-dimensional (3D) culture models have the potential to bridge the gap between monolayer cell culture and in vivo studies. To benefit anti-cancer drug discovery from 3D models, new techniques are needed that enable their use in high-throughput (HT) screening amenable formats. We have established miniaturized 3D culture methods robust enough for automated HT screens. We have applied these methods to evaluate the sensitivity of normal and tumorigenic breast epithelial cell lines against a panel of oncology drugs when cultured as monolayers (2D) and spheroids (3D). We have identified two classes of compounds that exhibit preferential cytotoxicity against cancer cells over normal cells when cultured as 3D spheroids: microtubule-targeting agents and epidermal growth factor receptor (EGFR) inhibitors. Further improving upon our 3D model, superior differentiation of EC50 values in the proof-of-concept screens was obtained by co-culturing the breast cancer cells with normal human fibroblasts and endothelial cells. Further, the selective sensitivity of the cancer cells towards chemotherapeutics was observed in 3D co-culture conditions, rather than as 2D co-culture monolayers, highlighting the importance of 3D cultures. Finally, we examined the putative mechanisms that drive the differing potency displayed by EGFR inhibitors. In summary, our studies establish robust 3D culture models of human cells for HT assessment of tumor cell-selective agents. This methodology is anticipated to provide a useful tool for the study of biological differences within 2D and 3D culture conditions in HT format, and an important platform for novel anti-cancer drug discovery.
authors.affiliation: Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America.
authors.forename: Amy L
authors.initials: AL
authors.lastname: Howes
authors.affiliation: Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America.
authors.forename: Robyn D
authors.initials: RD
authors.lastname: Richardson
authors.affiliation: Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America.
authors.forename: Darren
authors.initials: D
authors.lastname: Finlay
authors.affiliation: Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America.
authors.forename: Kristiina
authors.initials: K
authors.lastname: Vuori
mesh_terms: D000970:Antineoplastic Agents / Q000145:classification / Q000494:pharmacology* / Q000633:toxicity; D001940:Breast / Q000166:cytology*; D001943:Breast Neoplasms / Q000473:pathology*; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D018920:Coculture Techniques; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor / Q000295:instrumentation / Q000379:methods*; D004847:Epithelial Cells / Q000187:drug effects; D066246:ErbB Receptors / Q000037:antagonists & inhibitors*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects; D057166:High-Throughput Screening Assays; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008297:Male; D008904:Miniaturization; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology* / Q000633:toxicity; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D018874:Spheroids, Cellular / Q000187:drug effects; D050257:Tubulin Modulators / Q000494:pharmacology / Q000633:toxicity
chemical_list: D000970:Antineoplastic Agents; D066246:ErbB Receptors; D009363:Neoplasm Proteins; D047428:Protein Kinase Inhibitors; D018719:Erb-b2 Receptor Tyrosine Kinases; D050257:Tubulin Modulators; C512478:EGFR protein, human; C508053:ERBB2 protein, human