Screening for Small-Molecule Modulators of Long Noncoding RNA-Protein Interactions Using AlphaScreen

• Published in: Journal of biomolecular screening Vol. 20; no. 9; pp. 1132 - 1141
• Main Authors: Pedram Fatemi, Roya, Salah-Uddin, Sultan, Modarresi, Farzaneh, Khoury, Nathalie, Wahlestedt, Claes, Faghihi, Mohammad Ali
• Format: Journal Article
• Language: English
• Published: Los Angeles, CA SAGE Publications 01.10.2015
• Subjects: Brain-Derived Neurotrophic Factor - biosynthesis; Brain-Derived Neurotrophic Factor - genetics; Drug Evaluation, Preclinical; Ellipticines - pharmacology; Enhancer of Zeste Homolog 2 Protein; Gene Expression - drug effects; HEK293 Cells; High-Throughput Screening Assays; Humans; Polycomb Repressive Complex 2 - biosynthesis; Polycomb Repressive Complex 2 - genetics; Protein Binding - drug effects; RNA Interference - drug effects; RNA, Long Noncoding - antagonists & inhibitors; RNA, Long Noncoding - biosynthesis; RNA, Long Noncoding - chemistry; brain-derived neurotrophic factor antisense (BDNFAS); epigenetic enzyme; RNA protein interaction; noncoding RNA; hox transcript antisense RNA (HOTAIR); AlphaScreen; brain-derived neurotrophic factor (BDNF); long noncoding RNA; Enhancer of zeste homolog 2 (EZH2); natural antisense transcript
• ISSN: 1087-0571; 2472-5552; 1552-454X
• DOI: 10.1177/1087057115594187

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.