title: Large-scale pharmacological profiling of 3D tumor models of cancer cells.
abstract: The discovery of chemotherapeutic agents for the treatment of cancer commonly uses cell proliferation assays in which cells grow as two-dimensional (2D) monolayers. Compounds identified using 2D monolayer assays often fail to advance during clinical development, most likely because these assays do not reproduce the cellular complexity of tumors and their microenvironment in vivo. The use of three-dimensional (3D) cellular systems have been explored as enabling more predictive in vitro tumor models for drug discovery. To date, small-scale screens have demonstrated that pharmacological responses tend to differ between 2D and 3D cancer cell growth models. However, the limited scope of screens using 3D models has not provided a clear delineation of the cellular pathways and processes that differentially regulate cell survival and death in the different in vitro tumor models. Here we sought to further understand the differences in pharmacological responses between cancer tumor cells grown in different conditions by profiling a large collection of 1912 chemotherapeutic agents. We compared pharmacological responses obtained from cells cultured in traditional 2D monolayer conditions with those responses obtained from cells forming spheres versus cells already in 3D spheres. The target annotation of the compound library screened enabled the identification of those key cellular pathways and processes that when modulated by drugs induced cell death in all growth conditions or selectively in the different cell growth models. In addition, we also show that many of the compounds targeting these key cellular functions can be combined to produce synergistic cytotoxic effects, which in many cases differ in the magnitude of their synergism depending on the cellular model and cell type. The results from this work provide a high-throughput screening framework to profile the responses of drugs both as single agents and in pairwise combinations in 3D sphere models of cancer cells.
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Lesley A
authors.initials: LA
authors.lastname: Mathews Griner
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Xiaohu
authors.initials: X
authors.lastname: Zhang
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Rajarshi
authors.initials: R
authors.lastname: Guha
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Crystal
authors.initials: C
authors.lastname: McKnight
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Ian S
authors.initials: IS
authors.lastname: Goldlust
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Madhu
authors.initials: M
authors.lastname: Lal-Nag
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Kelli
authors.initials: K
authors.lastname: Wilson
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Sam
authors.initials: S
authors.lastname: Michael
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Steve
authors.initials: S
authors.lastname: Titus
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Shinn
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Craig J
authors.initials: CJ
authors.lastname: Thomas
authors.affiliation: National Center for Advancing Translational Sciences, Division of Pre-Clinical Innovation, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Marc
authors.initials: M
authors.lastname: Ferrer
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008954:Models, Biological; D018874:Spheroids, Cellular / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents