title: A high-throughput screen for Wnt/β-catenin signaling pathway modulators in human iPSC-derived neural progenitors.
abstract: Wnt/β-catenin signaling has emerged as a central player in pathways implicated in the pathophysiology and treatment of neuropsychiatric disorders. To identify potential novel therapeutics for these disorders, high-throughput screening (HTS) assays reporting on Wnt/β-catenin signaling in disease-relevant contexts are needed. The use of human patient-derived induced pluripotent stem cell (iPSC) models provides ideal disease-relevant context if these stem cell cultures can be adapted for HTS-compatible formats. Here, we describe a sensitive, HTS-compatible Wnt/β-catenin signaling reporter system generated in homogeneous, expandable neural progenitor cells (NPCs) derived from human iPSCs. We validated this system by demonstrating dose-responsive stimulation by several known Wnt/β-catenin signaling pathway modulators, including Wnt3a, a glycogen synthase kinase-3 (GSK3) inhibitor, and the bipolar disorder therapeutic lithium. These responses were robust and reproducible over time across many repeated assays. We then conducted a screen of ~1500 compounds from a library of Food and Drug Administration-approved drugs and known bioactives and confirmed the HTS hits, revealing multiple chemical and biological classes of novel small-molecule probes of Wnt/β-catenin signaling. Generating these type of pathway-selective, cell-based phenotypic assays in human iPSC-derived neural cells will advance the field of human experimental neurobiology toward the goal of identifying and validating targets for neuropsychiatric disorders.
authors.affiliation: Center for Human Genetic Research, Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA.
authors.forename: Wen-Ning
authors.initials: WN
authors.lastname: Zhao
authors.forename: Chialin
authors.initials: C
authors.lastname: Cheng
authors.forename: Kraig M
authors.initials: KM
authors.lastname: Theriault
authors.forename: Steven D
authors.initials: SD
authors.lastname: Sheridan
authors.forename: Li-Huei
authors.initials: LH
authors.lastname: Tsai
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Haggarty
mesh_terms: D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D008094:Lithium / Q000494:pharmacology; D001523:Mental Disorders / Q000188:drug therapy / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015203:Reproducibility of Results; D054852:Small Molecule Libraries / Q000494:pharmacology; D060449:Wnt Signaling Pathway / Q000187:drug effects*; D060509:Wnt3A Protein / Q000378:metabolism; D051176:beta Catenin / Q000378:metabolism
chemical_list: D054852:Small Molecule Libraries; D060509:Wnt3A Protein; D051176:beta Catenin; D008094:Lithium; D038362:Glycogen Synthase Kinase 3