title: A High-Throughput, Multi-Cell Phenotype Assay for the Identification of Novel Inhibitors of Chemotaxis/Migration.
abstract: Chemotaxis and cell migration are fundamental, universal eukaryotic processes essential for biological functions such as embryogenesis, immunity, cell renewal, and wound healing, as well as for pathogenesis of many diseases including cancer metastasis and chronic inflammation. To identify novel chemotaxis inhibitors as probes for mechanistic studies and leads for development of new therapeutics, we developed a unique, unbiased phenotypic chemotaxis-dependent Dictyostelium aggregation assay for high-throughput screening using rapid, laser-scanning cytometry. Under defined conditions, individual Dictyostelium secrete chemoattractants, migrate, and aggregate. Chemotaxis is quantified by laser-scanning cytometry with a GFP marker expressed only in cells after chemotaxis/multi-cell aggregation. We applied the assay to screen 1,280 known compounds in a 1536-well plate format and identified two chemotaxis inhibitors. The chemotaxis inhibitory activities of both compounds were confirmed in both Dictyostelium and in human neutrophils in a directed EZ-TAXIscan chemotaxis assay. The compounds were also shown to inhibit migration of two human cancer cell lines in monolayer scratch assays. This test screen demonstrated that the miniaturized assay is extremely suited for high-throughput screening of very large libraries of small molecules to identify novel classes of chemotaxis/migratory inhibitors for drug development and research tools for targeting chemotactic pathways universal to humans and other systems.
authors.affiliation: Institute for Translational Medicine, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian 350108, China.|Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, The National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Xin-Hua
authors.initials: XH
authors.lastname: Liao
authors.affiliation: Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, The National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Netra Pal
authors.initials: NP
authors.lastname: Meena
authors.affiliation: Therapeutics for Rare and Neglected Diseases, National Center for Advancing Translational Sciences, The National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Noel
authors.initials: N
authors.lastname: Southall
authors.affiliation: Laboratory of Cellular and Molecular Biology, National Cancer Institute, The National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Lunhua
authors.initials: L
authors.lastname: Liu
authors.affiliation: Therapeutics for Rare and Neglected Diseases, National Center for Advancing Translational Sciences, The National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Manju
authors.initials: M
authors.lastname: Swaroop
authors.affiliation: Institute for Translational Medicine, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian 350108, China.
authors.forename: Arina Li
authors.initials: AL
authors.lastname: Zhang
authors.affiliation: Institute for Translational Medicine, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian 350108, China.
authors.forename: Jan Jian
authors.initials: JJ
authors.lastname: Xiang
authors.affiliation: Laboratory of Cellular and Molecular Biology, National Cancer Institute, The National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Carole A
authors.initials: CA
authors.lastname: Parent
authors.affiliation: Therapeutics for Rare and Neglected Diseases, National Center for Advancing Translational Sciences, The National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Zheng
authors.affiliation: Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, The National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Alan R
authors.initials: AR
authors.lastname: Kimmel
mesh_terms: D002465:Cell Movement / Q000187:drug effects*; D002633:Chemotaxis / Q000187:drug effects*; D003603:Cytotoxins / Q000302:isolation & purification* / Q000494:pharmacology*; D004023:Dictyostelium / Q000187:drug effects* / Q000502:physiology; D004353:Drug Evaluation, Preclinical / Q000379:methods*
chemical_list: D003603:Cytotoxins