RNA sequencing in primary inflammatory (TPP) macrophages following deletion of a disease-associated gene desert at chr21q22, disruption of ETS2, or treatment of ETS2-edited macrophages with a HIF1α stabiliser.

GWAS studies in five different inflammatory diseases have identified a strong genetic association at a gene desert at the chr21q22 locus. We have shown that this locus contains a monocyte/macrophage-specific enhancer that regulates ETS2 - a gene whose role in primary human monocytes/macrophages is incompletely understood. We therefore used a CRISPR-Cas9-based approach to delete the enhancer region or to disrupt ETS2, and performed RNA-sequencing to examine the transcriptional consequences. One effect of ETS2 disruption was upregulation of genes involved in aerobic respiration and oxidative phosphorylation. We therefore treated ETS2-edited inflammatory macrophages with roxadustat, a HIF1α stabiliser that can promote glycolysis via HIF1α-mediated metabolic reprogramming, and performed RNA-sequencing to determine whether this drug might rescue the transcriptional effects of ETS2 disruption.EGA study EGAS00001007553

针对五种不同炎症性疾病开展的全基因组关联研究（Genome-Wide Association Study, GWAS），在21号染色体q22区域的基因荒漠区发现了显著的遗传关联。我们的研究证实，该位点存在一个单核细胞/巨噬细胞特异性增强子，可调控ETS2基因——该基因在原代人单核细胞/巨噬细胞中的功能尚未得到充分阐明。为此，我们采用基于CRISPR-Cas9的技术手段，敲除该增强子区域或破坏ETS2基因，并通过RNA测序（RNA-sequencing）分析其转录层面的调控效应。ETS2基因破坏所产生的效应之一，是需氧呼吸与氧化磷酸化相关基因的表达上调。据此，我们使用罗沙司他（roxadustat）——一种可通过HIF1α介导的代谢重编程促进糖酵解的HIF1α稳定剂——处理经CRISPR编辑的炎症性巨噬细胞，并再次开展RNA测序，以明确该药物是否能够挽救ETS2破坏引发的转录组改变。本研究的欧洲基因组-表型档案（European Genome-phenome Archive, EGA）存档编号为EGAS00001007553