title: High-Throughput Screening Using iPSC-Derived Neuronal Progenitors to Identify Compounds Counteracting Epigenetic Gene Silencing in Fragile X Syndrome.
abstract: Fragile X syndrome (FXS) is the most common form of inherited mental retardation, and it is caused in most of cases by epigenetic silencing of the Fmr1 gene. Today, no specific therapy exists for FXS, and current treatments are only directed to improve behavioral symptoms. Neuronal progenitors derived from FXS patient induced pluripotent stem cells (iPSCs) represent a unique model to study the disease and develop assays for large-scale drug discovery screens since they conserve the Fmr1 gene silenced within the disease context. We have established a high-content imaging assay to run a large-scale phenotypic screen aimed to identify compounds that reactivate the silenced Fmr1 gene. A set of 50,000 compounds was tested, including modulators of several epigenetic targets. We describe an integrated drug discovery model comprising iPSC generation, culture scale-up, and quality control and screening with a very sensitive high-content imaging assay assisted by single-cell image analysis and multiparametric data analysis based on machine learning algorithms. The screening identified several compounds that induced a weak expression of fragile X mental retardation protein (FMRP) and thus sets the basis for further large-scale screens to find candidate drugs or targets tackling the underlying mechanism of FXS with potential for therapeutic intervention.
authors.affiliation: Center of Proteomic Chemistry, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Markus
authors.initials: M
authors.lastname: Kaufmann
authors.affiliation: Center of Proteomic Chemistry, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Ansgar
authors.initials: A
authors.lastname: Schuffenhauer
authors.affiliation: Developmental and Molecular Pathways, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Fruh
authors.affiliation: Developmental and Molecular Pathways, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Jessica
authors.initials: J
authors.lastname: Klein
authors.affiliation: Developmental and Molecular Pathways, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Anke
authors.initials: A
authors.lastname: Thiemeyer
authors.affiliation: Center of Proteomic Chemistry, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Pierre
authors.initials: P
authors.lastname: Rigo
authors.affiliation: Translational Medicine, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Baltazar
authors.initials: B
authors.lastname: Gomez-Mancilla
authors.affiliation: Center of Proteomic Chemistry, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Valerie
authors.initials: V
authors.lastname: Heidinger-Millot
authors.affiliation: Developmental and Molecular Pathways, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Tewis
authors.initials: T
authors.lastname: Bouwmeester
authors.affiliation: Center of Proteomic Chemistry, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Schopfer
authors.affiliation: Developmental and Molecular Pathways, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Matthias
authors.initials: M
authors.lastname: Mueller
authors.affiliation: Developmental and Molecular Pathways, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Barna D
authors.initials: BD
authors.lastname: Fodor
authors.affiliation: Center of Proteomic Chemistry, Novartis Institutes for Biomedical Research, Novartis Campus, Basel, Switzerland amanda.cobos-correa@novartis.com.
authors.forename: Amanda
authors.initials: A
authors.lastname: Cobos-Correa
mesh_terms: D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000235:genetics / Q000378:metabolism; D005600:Fragile X Syndrome / Q000188:drug therapy* / Q000235:genetics; D020868:Gene Silencing / Q000187:drug effects*; D057166:High-Throughput Screening Assays; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000502:physiology; D058953:Neural Stem Cells / Q000187:drug effects* / Q000502:physiology; D018911:Trinucleotide Repeats
chemical_list: D051860:Fragile X Messenger Ribonucleoprotein 1; C497683:FMR1 protein, human
keywords: fragile X syndrome; high-content screening; induced pluripotent stem cells