Screening for inhibitors of orphan nuclear receptor DAX-1

描述（由申请人提供）：本研究的目的是通过 HTS 筛选程序鉴定孤儿核受体 DAX-1 (NR0B1) 的小分子抑制剂。 DAX-1 活性的抑制作为干细胞生物学和癌症领域的治疗工具具有潜在的意义。将通过简单的转染测定进行抑制剂筛选，使用含有 Gal4 结合位点的荧光素酶报告基因上的 Gal4 - DAX-1 C 末端结构域融合的转录抑制活性的减轻作为读数。 Countersreen 测定随后将涉及对在 DAX-1 转录特性的主要 HTS 活动期间鉴定的命中分子的活性的研究，这些分子是通过一组 DAX-1 响应报告基因（即类固醇生成酶基因）测量的。最后，三级测定将旨在开发具有改进的 DAX-1 抑制剂效力的化合物，评估它们对其他转录因子的作用选择性并测量它们的毒性。此外，还将测试抑制剂对 DAX-1 依赖性肿瘤细胞系生长的功效。 公共卫生相关性：开发具有更高选择性和更低毒性的新药对于现代抗癌治疗至关重要。我们的项目旨在寻找孤儿核受体 DAX-1 的新抑制剂，该受体在儿童和青少年特定类型软组织肿瘤的生长中发挥着关键作用。

DESCRIPTION (provided by applicant): The aim of this study will be to identify small-molecule inhibitors of orphan nuclear receptor DAX-1 (NR0B1) through a HTS screening program. Inhibition of DAX-1 activity has a potential interest as a therapeutic tool in the domains of stem cell biology and cancer. Inhibitor screening will be performed by a simple transfection assay using as readout the relief of the transcriptional repressor activity of a Gal4 - DAX-1 C-terminal domain fusion on a luciferase reporter harboring Gal4 binding sites. Countersreen assays will subsequently involve the study of the activity of the hit molecules identified during the primary HTS campaign on DAX-1 transcriptional properties measured on a battery of DAX-1 responsive reporters, i.e. steroidogenic enzyme genes. Finally, tertiary assays will be aimed at developing compounds with improved DAX-1 inhibitor potency, assess their selectivity of action against other transcription factors and measure their toxicity. Furthermore, the efficacy of the inhibitors will be tested on DAX-1 - dependent growth of tumor cell lines. PUBLIC HEALTH RELEVANCE: Development of new drugs displaying increased selectivity and less toxicity is of utmost importance for modern anticancer therapy. Our project has the aim to search for new inhibitors for the orphan nuclear receptor DAX-1, which has a pivotal role in the growth of a specific type of soft tissue tumor of children and adolescents.