Screening for inhibitors of orphan nuclear receptor DAX-1

描述(申请人提供)：这项研究的目的将是通过HTS筛选计划识别孤儿核受体DAX-1(NR0B1)的小分子抑制剂。抑制DAX-1活性作为干细胞生物学和癌症领域的治疗工具具有潜在的兴趣。抑制剂的筛选将通过一种简单的转染实验进行，该实验使用含有Gal4结合位点的荧光素酶报告上的Gal4-DAX-1 C末端结构域融合的转录抑制活性的解除作为读出。Countersreen分析随后将涉及在最初的HTS活动中确定的HIT分子在DAX-1上的活性的研究，该转录特性是在一组DAX-1响应报告上测量的，即类固醇合成酶基因。最后，三级检测的目的将是开发具有改进的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，这些抑制剂的有效性将在依赖DAX-1的肿瘤细胞系的生长上进行测试。 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现代抗癌治疗至关重要。我们的项目旨在为孤儿核受体DAX-1寻找新的抑制剂，该受体在儿童和青少年的一种特定类型的软组织肿瘤的生长中起着关键作用。

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.