Chemical probes targeting nuclear effector complex of the Hippo signaling pathway

针对 Hippo 信号通路核效应复合物的化学探针

• 批准号: 8791311
• 负责人: DUOJIA PAN
• 金额: $ 35.24万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8791311
• 关键词: Agar; Animals; Biochemical; Biological Assay; Biology; Cells; Chemicals; Co-Immunoprecipitations; Collaborations; Collection; Complex; DNA Binding; Development; Distal; Dose; Drosophila genus; Drug Kinetics; Elements; Epigenetic Process; Escherichia coli; Firefly Luciferases; Fluorescence Resonance Energy Transfer; Functional disorder; Gene Targeting; Genetic; Genetic Transcription; Genomics; Goals; Health; Homeostasis; Human; Hyperplasia; In Vitro; Lead; Libraries; Link; Liver; Luciferases; Malignant Neoplasms; Malignant neoplasm of liver; Mammals; Molecular; Molecular Bank; Mutation; Natural regeneration; Neurofibromatoses; Normal tissue morphology; Nuclear; Oncogene Proteins; Oncogenes; Oncogenic; Organ; Organ Size; Pathway interactions; Pharmaceutical Preparations; Phosphotransferases; Play; Property; Protein Kinase; Proteins; Reporter; Research; Robotics; Role; Scallop; Signal Pathway; Signal Transduction; System; Testing; Therapeutic; Tissues; Transcription Coactivator; Tumor Suppressor Genes; Tumor Suppressor Proteins; United States National Institutes of Health; Ursidae Family; Validation; base; cofactor; connective tissue growth factor; design; follow-up; high throughput screening; human disease; in vivo; inhibitor/antagonist; liver hyperplasia; miniaturize; mouse model; overexpression; repository; screening; small molecule; success; therapeutic development; therapeutic target; tool; transcription factor

DESCRIPTION (provided by applicant): Many organs such as the mammalian liver possess intrinsic information about their final size, yet the molecular mechanisms that govern organ size in animal development and regeneration remain poorly understood. The recent discovery of the Hippo signaling pathway provides an unprecedented entry point to resolving this long- standing puzzle in biology. First discovered in Drosophila, the Hippo signaling pathway is an evolutionarily conserved regulator of organ size. Central to the Hippo pathway is a kinase cascade leading from the protein kinase Hpo/Mst to a transcription factor complex formed between coactivator Yki/YAP and DNA-binding transcription factor Sd/TEAD. Consistent with the critical role of Hippo signaling in normal tissue homeostasis, the YAP protein is overexpressed or hyperactivated in a wide spectrum of human cancers due to YAP locus amplification or genetic/epigenetic inactivation of upstream tumor suppressors. Small molecule inhibitors of YAP will not only provide important tools for pharmacological manipulation of Hippo signaling, but also bear tremendous potential for developing therapeutic drugs against human diseases caused by defective Hippo signaling. To this end, we have designed and functionally validated a primary high-throughput screen assay to discover chemicals capable of inhibiting the interaction of coactivator Yki/YAP and its cognate transcription factor Sd/TEAD. We have collaborated the Chemical Genomics Branch at NIH Chemical Genomics Center (NCGC) to miniaturize this assay to 1536-plate format and successfully conducted a pilot screen with the Library of Pharmacologically Active Compounds 1280 (LOPAC1280). In this proposed research, we aim to first implement this target-based FRET assay in a high-throughput screen against the National Institute of Health- Molecular Libraries Small Molecule Repository (NIH-MLSMR) collection covering a wide range of chemical space at the NCGC screening center. Next, we will validate the primary screen hits using a plethora of pre- established follow-up assays to remove false positives and prioritize the hit set. Finally we will test the hits in two well-characterized Hippo pathway-specific mouse models of YAP-induced liver hyperplasia and liver cancer. Taken together, our proposed research represents the first-of-its-kind large-scale screening campaign to discover lead compounds targeting the Hippo signaling pathway.

描述（由申请人提供）：许多器官（例如哺乳动物肝脏）具有有关其最终大小的固有信息，但是控制动物发育和再生中器官大小的分子机制仍然很少了解。最近发现河马信号通路的发现提供了一个前所未有的入口点，以解决生物学中的长期拼图。河马信号通路最初是在果蝇中发现的，是器官大小的进化保守调节剂。河马途径的中心是从蛋白激酶HPO/MST到转录因子复合物YKI/YAP和DNA结合转录因子SD/TEAD之间形成的转录因子复合物SD/TEAD之间形成的转录因子复合物。与HIPPO信号在正常组织稳态中的关键作用一致，由于YAP基因座放大或上游肿瘤抑制剂的遗传学/表观遗传失活，YAP蛋白在广泛的人类癌症中过表达或过度活化。 YAP的小分子抑制剂不仅可以为河马信号的药理学操纵提供重要的工具，而且还具有巨大的潜力，可以开发由河马信号不良引起的人类疾病的治疗药物。为此，我们设计并在功能上验证了主要的高通量屏幕测定法，以发现能够抑制共激活因子YKI/YAP相互作用的化学物质及其认知转录因子SD/TEAD。我们已经合作了NIH化学基因组学中心（NCGC）的化学基因组学分支，以使该测定法至1536板格式，并与药理学活性化合物图书馆1280（LOPAC1280）一起成功地进行了试验筛选。在这项拟议的研究中，我们的目标是在针对国家卫生分子图书馆小分子存储库（NIH-MLSMR）集合中首先实施此基于目标的FRET测定法，该屏幕库中涵盖了NCGC筛查中心的各种化学空间。接下来，我们将使用大量预先确定的后续测定法验证主屏幕命中，以删除误报并确定命中率。最后，我们将测试两个特征良好的命中 YAP诱导的肝增生和肝癌的河马途径特异性小鼠模型。综上所述，我们提出的研究代表了首个大规模筛查运动，以发现针对河马信号通路的铅化合物。