title: Screening of Intestinal Crypt Organoids: A Simple Readout for Complex Biology.
abstract: Oral and intestinal mucositis is a debilitating side effect of radiation treatment. A mouse model of radiation-induced mucositis leads to weight loss and tissue damage, reflecting the human ailment as it responds to keratinocyte growth factor (KGF), the standard-of-care treatment. Cultured intestinal crypt organoids allowed the development of an assay monitoring the effect of treatments of intestinal epithelium to radiation-induced damage. This in vitro assay resembles the mouse model as KGF and roof plate-specific spondin-1 (RSPO1) enhanced crypt organoid recovery following radiation. Screening identified compounds that increased the survival of organoids postradiation. Testing of these compounds revealed that the organoids changed their responses over time. Unbiased transcriptome analysis was performed on crypt organoid cultures at various time points in culture to investigate this adaptive behavior. A number of genes and pathways were found to be modulated over time, providing a rationale for the altered sensitivity of the organoid cultures. This report describes an in vitro assay that reflects aspects of human disease. The assay was used to identify bioactive compounds, which served as probes to interrogate the biology of crypt organoids over prolonged culture. The pathways that are changing over time may offer potential targets for treatment of mucositis.
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Svenja
authors.initials: S
authors.lastname: Ley
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Olaf
authors.initials: O
authors.lastname: Galuba
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Adrian
authors.initials: A
authors.lastname: Salathe
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Melin
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Aebi
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Monika
authors.initials: M
authors.lastname: Pikiolek
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Judith
authors.initials: J
authors.lastname: Knehr
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Walter
authors.initials: W
authors.lastname: Carbone
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Beibel
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Florian
authors.initials: F
authors.lastname: Nigsch
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Guglielmo
authors.initials: G
authors.lastname: Roma
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Giovanni
authors.initials: G
authors.lastname: d'Ario
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Susan
authors.initials: S
authors.lastname: Kirkland
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Laure C
authors.initials: LC
authors.lastname: Bouchez
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Caroline
authors.initials: C
authors.lastname: Gubser Keller
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Tewis
authors.initials: T
authors.lastname: Bouwmeester
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Christian N
authors.initials: CN
authors.lastname: Parker
authors.affiliation: 1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
authors.forename: Heinz
authors.initials: H
authors.lastname: Ruffner
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D051523:Fibroblast Growth Factor 7 / Q000378:metabolism; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism; D007422:Intestines / Q000187:drug effects*; D051379:Mice; D008810:Mice, Inbred C57BL; D009940:Organoids / Q000187:drug effects* / Q000378:metabolism; D019699:Thrombospondins / Q000378:metabolism; D059467:Transcriptome / Q000502:physiology
chemical_list: D019699:Thrombospondins; D051523:Fibroblast Growth Factor 7
keywords: KGF; RSPO; mucositis; primary 3D cell cultures; small intestinal crypt organoids