title: Screening for Small-Molecule Modulators of Long Noncoding RNA-Protein Interactions Using AlphaScreen.
abstract: Long non-protein coding RNAs (lncRNAs) are an important class of molecules that help orchestrate key cellular events. Although their functional roles in cells are not well understood, thousands of lncRNAs and a number of possible mechanisms by which they act have been reported. LncRNAs can exert their regulatory function in cells by interacting with epigenetic enzymes. In this study, we developed a tool to study lncRNA-protein interactions for high-throughput screening of small-molecule modulators using AlphaScreen technology. We tested the interaction of two lncRNAs: brain-derived neurotrophic factor antisense (BDNF-AS) and Hox transcript antisense RNA (HOTAIR), with Enhancer of zeste homolog 2 (EZH2), a histone methyltransferase against a phytochemical library, to look for small-molecule inhibitors that can alter the expression of downstream target genes. We identified ellipticine, a compound that up-regulates BDNF transcription. Our study shows the feasibility of using high-throughput screening to identify modulators of lncRNA-protein interactions and paves the road for targeting lncRNAs that are dysregulated in human disorders using small-molecule therapies.
authors.affiliation: Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, USA Department of Psychiatry and Behavioral Sciences and Center for Therapeutic Innovation, University of Miami Miller School of Medicine, Miami, FL, USA.
authors.forename: Roya
authors.initials: R
authors.lastname: Pedram Fatemi
authors.affiliation: Department of Psychiatry and Behavioral Sciences and Center for Therapeutic Innovation, University of Miami Miller School of Medicine, Miami, FL, USA.
authors.forename: Sultan
authors.initials: S
authors.lastname: Salah-Uddin
authors.affiliation: Department of Psychiatry and Behavioral Sciences and Center for Therapeutic Innovation, University of Miami Miller School of Medicine, Miami, FL, USA.
authors.forename: Farzaneh
authors.initials: F
authors.lastname: Modarresi
authors.affiliation: Department of Psychiatry and Behavioral Sciences and Center for Therapeutic Innovation, University of Miami Miller School of Medicine, Miami, FL, USA.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Khoury
authors.affiliation: Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, USA Department of Psychiatry and Behavioral Sciences and Center for Therapeutic Innovation, University of Miami Miller School of Medicine, Miami, FL, USA.
authors.forename: Claes
authors.initials: C
authors.lastname: Wahlestedt
authors.affiliation: Department of Psychiatry and Behavioral Sciences and Center for Therapeutic Innovation, University of Miami Miller School of Medicine, Miami, FL, USA Department of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine, Miami, FL, USA mfaghihi@med.miami.edu.
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Faghihi
mesh_terms: D019208:Brain-Derived Neurotrophic Factor / Q000096:biosynthesis / Q000235:genetics; D004353:Drug Evaluation, Preclinical; D004611:Ellipticines / Q000494:pharmacology; D000071221:Enhancer of Zeste Homolog 2 Protein; D015870:Gene Expression / Q000187:drug effects; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D063151:Polycomb Repressive Complex 2 / Q000096:biosynthesis / Q000235:genetics; D011485:Protein Binding / Q000187:drug effects; D034622:RNA Interference / Q000187:drug effects; D062085:RNA, Long Noncoding / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000737:chemistry
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D004611:Ellipticines; C561566:HOTAIR long untranslated RNA, human; D062085:RNA, Long Noncoding; C034192:ellipticine; C579455:BDNF protein, human; C498409:EZH2 protein, human; D000071221:Enhancer of Zeste Homolog 2 Protein; D063151:Polycomb Repressive Complex 2
keywords: AlphaScreen; Enhancer of zeste homolog 2 (EZH2); RNA protein interaction; brain-derived neurotrophic factor (BDNF); brain-derived neurotrophic factor antisense (BDNFAS); epigenetic enzyme; hox transcript antisense RNA (HOTAIR); long noncoding RNA; natural antisense transcript; noncoding RNA