Inderbinen Silvia G, Kley Manuel, Zogg Michael, Sellner Manuel, Fischer André, Kędzierski Jacek, Boudon Stéphanie, Jetten Anton M, Smieško Martin, Odermatt Alex
Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, Klingelbergstrasse 50, 4056 Basel, Switzerland; Swiss Centre for Applied Human Toxicology and Department of Pharmaceutical Sciences, University of Basel, Missionsstrasse 64, 4055 Basel, Switzerland.
Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, Klingelbergstrasse 50, 4056 Basel, Switzerland.
Toxicology. 2022 Apr 15;471:153159. doi: 10.1016/j.tox.2022.153159. Epub 2022 Mar 23.
Retinoic acid-related orphan receptor γt (RORγt) regulates immune responses and its impaired function contributes to inflammatory and autoimmune diseases and may promote skin cancer. Synthetic inverse RORγt agonists block the production of Th17-associated cytokines including interleukin (IL)-17A and IL-22 and are under investigation for treatment of such pathologies. Unintentional RORγt activation in skin, following exposure to environmental chemicals, may promote inflammatory skin disease. Parabens and UV-filters, frequently used as additives in cosmetics and body care products, are intensively inspected for endocrine disrupting properties. This study assessed whether such compounds can interfere with RORγ activity using a previously established tetracycline-inducible reporter gene assay in CHO cells. These transactivation experiments revealed hexylparaben, benzylparaben and benzophenone-10 as RORγ agonists (EC values: 144 ± 97 nM, 3.39 ± 1.74 µM and 1.67 ± 1.04 µM, respectively), and they could restore RORγ activity after suppression by an inverse agonist. Furthermore, they enhanced RORγt-dependent transcription of the pro-inflammatory IL-17A and/or IL-22 genes in the murine T-cell model EL4. Virtual screening of a cosmetics database for structurally similar chemicals and in vitro testing of the most promising hits revealed benzylbenzoate, benzylsalicylate and 4-methylphenylbenzoate as RORγ agonists (low micromolar EC values). Moreover, an analysis of mixtures of the newly identified RORγ agonists suggested additive effects. This study presents novel RORγ(t) agonistic structural scaffolds. By activating RORγ(t) the identified parabens and UV-filters may potentially aggravate pathophysiological conditions, especially skin diseases where highest exposure of such chemicals can be expected. Follow-up studies should assess whether such compounds, either alone or as mixtures, can reach relevant concentrations in tissues and target cells to activate RORγ(t) in vivo.
维甲酸相关孤儿受体γt（RORγt）调节免疫反应，其功能受损会导致炎症性和自身免疫性疾病，并可能促进皮肤癌。合成型反向RORγt激动剂可阻断包括白细胞介素（IL）-17A和IL-22在内的Th17相关细胞因子的产生，目前正在研究其用于治疗此类病症。接触环境化学物质后皮肤中RORγt的意外激活可能会促进炎症性皮肤病。对羟基苯甲酸酯和紫外线过滤剂常用于化妆品和个人护理产品中，因其具有内分泌干扰特性而受到严格检查。本研究使用先前建立的CHO细胞四环素诱导报告基因测定法评估了此类化合物是否会干扰RORγ活性。这些反式激活实验表明，对羟基苯甲酸己酯、对羟基苯甲酸苄酯和二苯甲酮-10为RORγ激动剂（EC值分别为：144±97 nM、3.39±1.74 μM和1.67±1.04 μM），并且它们可以在被反向激动剂抑制后恢复RORγ活性。此外，它们增强了小鼠T细胞模型EL4中促炎IL-17A和/或IL-22基因的RORγt依赖性转录。对化妆品数据库进行结构相似化学物质的虚拟筛选，并对最有前景的命中物进行体外测试，结果表明苯甲酸苄酯、水杨酸苄酯和4-甲基苯基苯甲酸酯为RORγ激动剂（低微摩尔EC值）。此外，对新鉴定的RORγ激动剂混合物的分析表明存在加和效应。本研究提出了新型RORγ（t）激动剂结构支架。通过激活RORγ（t），已鉴定的对羟基苯甲酸酯和紫外线过滤剂可能会加剧病理生理状况，尤其是在预期此类化学物质暴露量最高的皮肤病中。后续研究应评估此类化合物单独或作为混合物是否能在组织和靶细胞中达到相关浓度以在体内激活RORγ（t）。
