title: A novel quantitative high-throughput screen identifies drugs that both activate SUMO conjugation via the inhibition of microRNAs 182 and 183 and facilitate neuroprotection in a model of oxygen and glucose deprivation.
abstract: The conjugation/de-conjugation of Small Ubiquitin-like Modifier (SUMO) has been shown to be associated with a diverse set of physiologic/pathologic conditions. The clinical significance and ostensible therapeutic utility offered via the selective control of the global SUMOylation process has become readily apparent in ischemic pathophysiology. Herein, we describe the development of a novel quantitative high-throughput screening (qHTS) system designed to identify small molecules capable of increasing SUMOylation via the regulation/inhibition of members of the microRNA (miRNA)-182 family. This assay employs a SHSY5Y human neuroblastoma cell line stably transfected with a dual firefly-Renilla luciferase reporter system for identification of specific inhibitors of either miR-182 or miR-183. In this study, we have identified small molecules capable of inducing increased global conjugation of SUMO in both SHSY5Y cells and rat E18-derived primary cortical neurons. The protective effects of a number of the identified compounds were confirmed via an in vitro ischemic model (oxygen/glucose deprivation). Of note, this assay can be easily repurposed to allow high-throughput analyses of the potential drugability of other relevant miRNA(s) in ischemic pathobiology.
authors.affiliation: Stroke Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health (NINDS/NIH), Bethesda, MD, USA Department of Clinical Neurosciences, Division of Stem Cell Neurobiology, Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Bernstock
authors.affiliation: Stroke Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health (NINDS/NIH), Bethesda, MD, USA.
authors.forename: Yang-ja
authors.initials: YJ
authors.lastname: Lee
authors.affiliation: Department of Clinical Neurosciences, Division of Stem Cell Neurobiology, Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.
authors.forename: Luca
authors.initials: L
authors.lastname: Peruzzotti-Jametti
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health (NCATS/NIH), Bethesda, MD, USA.
authors.forename: Noel
authors.initials: N
authors.lastname: Southall
authors.affiliation: Bioinformatics Section, Information Technology & Bioinformatics Program, Division of Intramural Research (DIR), (NINDS/NIH), Bethesda, MD, USA.
authors.forename: Kory R
authors.initials: KR
authors.lastname: Johnson
authors.affiliation: Flow Cytometry Core Facility, National Institute of Neurological Disorders and Stroke, National Institutes of Health (NINDS/NIH), Bethesda, MD, USA.
authors.forename: Dragan
authors.initials: D
authors.lastname: Maric
authors.affiliation: Department of Clinical Neurosciences, Division of Stem Cell Neurobiology, Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.
authors.forename: Giulio
authors.initials: G
authors.lastname: Volpe
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health (NCATS/NIH), Bethesda, MD, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Kouznetsova
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health (NCATS/NIH), Bethesda, MD, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Zheng
authors.affiliation: Department of Clinical Neurosciences, Division of Stem Cell Neurobiology, Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.
authors.forename: Stefano
authors.initials: S
authors.lastname: Pluchino
authors.affiliation: Stroke Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health (NINDS/NIH), Bethesda, MD, USA HallenbJ@ninds.nih.gov.
authors.forename: John M
authors.initials: JM
authors.lastname: Hallenbeck
mesh_terms: D000818:Animals; D002540:Cerebral Cortex / Q000187:drug effects; D003227:Conjugation, Genetic / Q000187:drug effects; D003712:Dendrites / Q000187:drug effects; D049229:Dendritic Spines / Q000187:drug effects; D005947:Glucose / Q000172:deficiency*; D057166:High-Throughput Screening Assays; D006801:Humans; D000860:Hypoxia / Q000473:pathology*; D035683:MicroRNAs / Q000187:drug effects*; D008959:Models, Neurological; D009474:Neurons / Q000187:drug effects; D018696:Neuroprotective Agents / Q000494:pharmacology*; D061251:Primary Cell Culture; D051381:Rats; D054852:Small Molecule Libraries; D025841:Small Ubiquitin-Related Modifier Proteins / Q000187:drug effects; D058207:Sumoylation / Q000187:drug effects*; D057170:Translational Research, Biomedical / Q000379:methods
chemical_list: C533449:MIRN183 microRNA, human; D035683:MicroRNAs; C538984:Mirn182 microRNA, human; D018696:Neuroprotective Agents; D054852:Small Molecule Libraries; D025841:Small Ubiquitin-Related Modifier Proteins; D005947:Glucose
keywords: High-throughput assay development; SUMO conjugation; miRNA; neuroprotection; translational research