Regulation of IL12B Expression in Human Macrophages by TALEN-mediated Epigenome Editing
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Regulation of IL12B Expression in Human Macrophages by TALENmediated Epigenome Editing* Meng CHEN1, 2, Hua ZHU1, 2, Yu-juan MAO1, 2, Nan CAO1, 2, Ya-li YU1, 2, Lian-yun LI3, Qiu ZHAO1, 2, Min WU3, Mei YE3# 1 Department of Gastroenterology, Zhongnan Hospital, Wuhan University, Wuhan 430071, China 2 Hubei Clinical Center and Key Lab of Intestinal and Colorectal Diseases, Wuhan 430071, China 3 College of Life Sciences, Wuhan University, Wuhan 430072, China Huazhong University of Science and Technology 2020
Summary: Although the exact etiology of inflammatory bowel disease (IBD) remains unclear, exaggerated immune response in genetically predisposed individuals has been reported. Th1 and Th17 cells mediate IBD development. Macrophages produce IL-12 and IL-23 that share p40 subunit encoded by IL12B gene as heteromer partner to drive Th1 and Th17 differentiation. The available animal and human data strongly support the pathogenic role of IL-12/IL-23 in IBD development and suggest that blocking p40 might be the potential strategy for IBD treatment. Furthermore, aberrant alteration of some cytokines expression via epigenetic mechanisms is involved in pathogenesis of IBD. In this study, we analyzed core promoter region of IL12B gene and investigated whether IL12B expression could be regulated through targeted epigenetic modification with gene editing technology. Transcription activator-like effectors (TALEs) are widely used in the field of genome editing and can specifically target DNA sequence in the host genome. We synthesized the TALE DNA-binding domains that target the promoter of human IL12B gene and fused it with the functional catalytic domains of epigenetic enzymes. Transient expression of these engineered enzymes demonstrated that the TALE-DNMT3A targeted the selected IL12B promoter region, induced loci-specific DNA methylation, and down-regulated IL-12B expression in various human cell lines. Collectively, our data suggested that epigenetic editing of IL12B through methylating DNA on its promoter might be developed as a potential therapeutic strategy for IBD treatment. Key words: transcription activator-like effectors; epigenome editing; DNA methylation; cytokine; IL12B; inflammatory bowel disease
Inflammatory bowel disease (IBD) is a chronic inflammatory condition of the gastrointestinal tract encompassing two major forms: Crohn’s disease (CD) and ulcerative colitis (UC). Although the etiology remains to be elucidated, accumulating evidence suggests immune cells, such as macrophages and CD4+ T cells, particular Th1 and Th17 cells are closely associated with development of IBD[1–3]. Macrophages can kill extraneous pathogens by phagocytosis and innate immune response; on the other hand, as a kind of antigen presenting cell (APC), macrophages can drive and strengthen adaptive immune response through secreting proinflammatory cytokines including IL-12 and IL-23[2]. In addition, CD is mainly mediated by Th1 and Th17 cells[4, 5]. Differentiated Th1 and Th17 cells produce a large number of cytokines which in Meng
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