RORα controls inflammatory state of human macrophages

• Published in: PloS one 2018-11, Vol.13 (11), p.e0207374-e0207374
• Main Authors: Nejati Moharrami, Neda, Bjørkøy Tande, Erlend, Ryan, Liv, Espevik, Terje, Boyartchuk, Victor
• Format: Article
• Language: English
• Subjects: Animal models; Biology and Life Sciences; Cell death; Clonal deletion; CRISPR; Cytokines; Cytokines - genetics; Cytokines - immunology; Deoxyribonucleic acid; DNA; Endoplasmic reticulum; Gene Deletion; Gene expression; Gene Expression Regulation - drug effects; Gene Expression Regulation - immunology; Genes; Genomes; Health sciences; HEK293 Cells; Humans; IL-1β; Inflammation; Inflammation - chemically induced; Inflammation - genetics; Inflammation - pathology; Interleukin 6; Ligands; Lipopolysaccharides; Lipopolysaccharides - toxicity; Macrophages; Macrophages - immunology; Macrophages - pathology; Medicine; Medicine and Health Sciences; Metabolism; Monocytes; NF-κB protein; Nuclear Receptor Subfamily 1, Group F, Member 1 - genetics; Nuclear Receptor Subfamily 1, Group F, Member 1 - immunology; Penicillin; Physiology; Protein Biosynthesis - drug effects; Protein Biosynthesis - immunology; Stimulation; THP-1 Cells; Transcription factors; Transcription, Genetic - drug effects; Transcription, Genetic - immunology; Tumor necrosis factor
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0207374

ROR family of nuclear receptor transcription factors forms nodes connecting metabolic and inflammatory signaling pathways. The RORα members of the family have intrinsic transcriptional activity and they are involved in both activation and repression of a wide range of genes. The role of RORα in control of inflammation has been extensively studied using animal models but its function in human cells is not as well understood. To address this shortcoming, we analyzed how RORα is shaping the inflammatory state of human macrophages. Using CRISPR-Cas9 system, we deleted RORA in THP-1 human monocytic cell line. In mutant cells we observed a dramatic increase in basal expression of a subset of NF-κB regulated genes, including TNF, IL-1β and IL-6, at both transcriptional and translational levels. Furthermore, RORA-deletion cells produced notable amounts of pro-IL-1β even in the absence of LPS stimulation. Subsequent LPS stimulation induced cleavage of pro-IL-1β to mature form. Our RNAseq analysis further confirmed the key role of RORA in setting the inflammatory state of macrophages and defined the set of differentially regulated genes. Overall, our data provides evidence supporting the anti-inflammatory function of RORα in human macrophages.