title: Assay Establishment and Validation of a High-Throughput Screening Platform for Three-Dimensional Patient-Derived Colon Cancer Organoid Cultures.
abstract: The application of patient-derived three-dimensional culture systems as disease-specific drug sensitivity models has enormous potential to connect compound screening and clinical trials. However, the implementation of complex cell-based assay systems in drug discovery requires reliable and robust screening platforms. Here we describe the establishment of an automated platform in 384-well format for three-dimensional organoid cultures derived from colon cancer patients. Single cells were embedded in an extracellular matrix by an automated workflow and subsequently self-organized into organoid structures within 4 days of culture before being exposed to compound treatment. We performed validation of assay robustness and reproducibility via plate uniformity and replicate-experiment studies. After assay optimization, the patient-derived organoid platform passed all relevant validation criteria. In addition, we introduced a streamlined plate uniformity study to evaluate patient-derived colon cancer samples from different donors. Our results demonstrate the feasibility of using patient-derived tumor samples for high-throughput assays and their integration as disease-specific models in drug discovery.
authors.affiliation: Eli Lilly and Company, Lilly Research Laboratories, Quantitative Biology, Alcobendas, Madrid, Spain.
authors.forename: Karsten
authors.initials: K
authors.lastname: Boehnke
authors.affiliation: Eli Lilly and Company, Lilly Research Laboratories, Global Discovery Statistics, Lilly Corporate Center, Indianapolis, IN, USA.
authors.forename: Philip W
authors.initials: PW
authors.lastname: Iversen
authors.affiliation: Institute of Pathology, Charité-Universitätsmedizin Berlin, Berlin, Germany German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Schumacher
authors.affiliation: Eli Lilly and Company, Lilly Research Laboratories, Quantitative Biology, Alcobendas, Madrid, Spain.
authors.forename: María José
authors.initials: MJ
authors.lastname: Lallena
authors.affiliation: Eli Lilly and Company, Lilly Research Laboratories, Discovery Chemistry Research & Technologies, Alcobendas, Madrid, Spain.
authors.forename: Rubén
authors.initials: R
authors.lastname: Haro
authors.affiliation: Eli Lilly and Company, Lilly Research Laboratories, Quantitative Biology, Alcobendas, Madrid, Spain.
authors.forename: Joaquín
authors.initials: J
authors.lastname: Amat
authors.affiliation: Institute of Pathology, Medical University Graz, Graz, Austria.
authors.forename: Johannes
authors.initials: J
authors.lastname: Haybaeck
authors.affiliation: German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Heidelberg, Germany Charité Comprehensive Cancer Center, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Liebs
authors.affiliation: Bayer Pharma AG, Berlin, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Lange
authors.affiliation: Institute of Pathology, Charité-Universitätsmedizin Berlin, Berlin, Germany German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Reinhold
authors.initials: R
authors.lastname: Schäfer
authors.affiliation: Institute of Pathology, Charité-Universitätsmedizin Berlin, Berlin, Germany CPO - Cellular Phenomics & Oncology, Berlin-Buch GmbH, Berlin, Germany.
authors.forename: Christian R A
authors.initials: CR
authors.lastname: Regenbrecht
authors.affiliation: Eli Lilly and Company, Lilly Research Laboratories, Oncology Translational Research, Lilly Corporate Center, Indianapolis, IN, USA.
authors.forename: Christoph
authors.initials: C
authors.lastname: Reinhard
authors.affiliation: Eli Lilly and Company, Lilly Research Laboratories, Quantitative Biology, Alcobendas, Madrid, Spain jvelasco@lilly.com.
authors.forename: Juan A
authors.initials: JA
authors.lastname: Velasco
mesh_terms: D000970:Antineoplastic Agents / Q000302:isolation & purification* / Q000627:therapeutic use; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000188:drug therapy; D055808:Drug Discovery / Q000379:methods; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D009940:Organoids / Q000254:growth & development* / Q000473:pathology; D018874:Spheroids, Cellular / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents
keywords: assay validation; cell-based assays; oncology; patient-derived organoid culture