Functional variants of the gene for the cytokine macrophage migration inhibitory factor (MIF) are defined by a 4-nucleotide promoter microsatellite (−794 CATT, rs5844572) and confer risk for autoimmune, infectious, and oncologic diseases. We describe herein the discovery of a prototypic, small molecule inhibitor of transcription with selectivity for high microsatellite repeat number and correspondingly high gene expression. Utilizing a high-throughput luminescent proximity screen, we identify 1-carbomethoxy-5-formyl-4,6,8-trihydroxyphenazine (CMFT) to inhibit the functional interaction between the transcription factor ICBP90 (, UHRF1) and the -794 CATT promoter microsatellite. CMFT inhibits mRNA expression in a −794 CATT length-dependent manner with an IC of 470 nM, and preferentially reduces ICBP90-dependent MIF mRNA and protein expression in high-genotypic low-genotypic expressing macrophages. RNA expression analysis also showed CMFT to downregulate MIF-dependent, inflammatory gene expression with little evidence of off-target metabolic toxicity. These findings provide proof-of-concept for advancing the pharmacogenomic development of precision-based MIF inhibitors for diverse autoimmune and inflammatory conditions.

中文翻译：

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MIF 免疫易感位点的小分子等位基因选择性转录抑制剂


细胞因子巨噬细胞迁移抑制因子 (MIF) 基因的功能变异由 4 核苷酸启动子微卫星（−794 CATT，rs5844572）定义，并赋予自身免疫性疾病、传染病和肿瘤疾病的风险。我们在此描述了对高微卫星重复数和相应高基因表达具有选择性的原型小分子转录抑制剂的发现。利用高通量发光邻近屏幕，我们鉴定出 1-甲氧基-5-甲酰基-4,6,8-三羟基吩嗪 (CMFT) 可以抑制转录因子 ICBP90（UHRF1）和 -794 CATT 启动子微卫星之间的功能相互作用。 CMFT 以 -794 CATT 长度依赖性方式抑制 mRNA 表达，IC 为 470 nM，并优先降低高基因型低基因型表达巨噬细胞中 ICBP90 依赖性 MIF mRNA 和蛋白质表达。 RNA 表达分析还显示 CMFT 下调 MIF 依赖性炎症基因表达，几乎没有脱靶代谢毒性的证据。这些发现为推进针对多种自身免疫和炎症性疾病的基于精准的 MIF 抑制剂的药物基因组学开发提供了概念验证。