Super-enhancers for RUNX3 are required for cell proliferation in EBV-infected B cell lines

• Published in: Gene Vol. 774; p. 145421
• Main Authors: Hosoi, Hiroki, Niibori-Nambu, Akiko, Nah, Giselle Sek Suan, Bahirvani, Avinash Govind, Mok, Michelle Meng Huang, Sanda, Takaomi, Kumar, Alan Prem, Tenen, Daniel G., Ito, Yoshiaki, Sonoki, Takashi, Osato, Motomi
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 30.03.2021
• Subjects: Azepines - pharmacology; B-Lymphocytes - cytology; B-Lymphocytes - virology; Burkitt Lymphoma - genetics; Burkitt Lymphoma - virology; Cell Line; Cell Line, Tumor; Cell Proliferation - drug effects; Cell Proliferation - genetics; Core Binding Factor Alpha 3 Subunit - genetics; CRISPR-Cas Systems; Enhancer Elements, Genetic; Epstein-Barr virus; G1 Phase - drug effects; Gene Expression Regulation - drug effects; Genes, myc; Herpesvirus 4, Human - physiology; Humans; JQ1; MYC; Protein Domains; Proteins - antagonists & inhibitors; Proteins - metabolism; RUNX3; Super-enhancer; Triazoles - pharmacology; CRISPR/Cas9; seMyc; seR3; qRT-PCR; EBNA2; Super-enhancer; siRNA; JQ1; RUNX3; eRNA; ECL; SDS-PAGE; LCL; sgRNA; TSS; MYC; ChIP-seq; EBV; PI; Epstein-Barr virus; T-ALL; gDNA; NK/T cell
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2021.145421

•Pharmacological inhibition on seR3s suppresses EBV-positive B cell proliferation.•CRISPR/Cas9-medicated excision of seR3s reduces propagation of the cells.•seR3s may function in trans on seMYC. Epstein-Barr virus nuclear antigens 2 (EBNA2) mediated super-enhancers, defined by in silico data, localize near genes associated with B cell transcription factors including RUNX3. However, the biological function of super-enhancer for RUNX3 gene (seR3) remains unclear. Here, we show that two seR3s, tandemly-located at 59- and 70-kb upstream of RUNX3 transcription start site, named seR3 −59h and seR3 −70h, are required for RUNX3 expression and cell proliferation in Epstein-Barr virus (EBV)-positive malignant B cells. A BET bromodomain inhibitor, JQ1, potently suppressed EBV-positive B cell growth through the reduction of RUNX3 and MYC expression. Excision of either or both seR3s by employing CRISPR/Cas9 system resulted in the decrease in RUNX3 expression and the subsequent suppression of cell proliferation and colony forming capability. The expression of MYC was also reduced when seR3s were deleted, probably due to the loss of trans effect of seR3s on the super-enhancers for MYC. These findings suggest that seR3s play a pivotal role in expression and biological function of both RUNX3 and MYC. seR3s would serve as a potential therapeutic target in EBV-related widespread tumors.