A high content screen dissecting ciliogenesis and oncogenic Hedgehog signaling

高内涵筛选剖析纤毛发生和致癌 Hedgehog 信号传导

基本信息

批准号：
9107196
负责人：
Michelle Arkin
金额：
$ 36.26万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-23 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9107196
关键词：
Alleles Allosteric Site Basal Cell Nevus Syndrome Basal cell carcinoma Behavior Binding Biological Assay Biological Process Biology Cancer Etiology Cell Cycle Progression Cell Maintenance Cells Chemicals Childhood Solid Neoplasm Cilia Cilium Microtubule Clinical Combination Drug Therapy Complex Critical Pathways Development Disease Dissection Distal Embryonic Development Erinaceidae Familial disease Genetic Genetic Transcription Growth Housing Human Integral Membrane Protein Investigation Lead Learning Libraries Ligands Malignant Childhood Neoplasm Malignant Epithelial Cell Malignant Neoplasms Mammalian Cell Microtubules Molecular Movement Mutation Natural Products Oncogenic Organelles Pathway interactions Pattern Pharmaceutical Preparations Predisposition Process Proteins Reagent Relapse Resistance Signal Transduction Signaling Protein Site Solid Stem cells Surface Testing Tissues Translations United States United States National Institutes of Health Vertebrates Vesnarinone appendage base cell growth cilium biogenesis combat counterscreen cyclopamine drug development fighting hedgehog signal transduction inhibitor/antagonist insight kinetosome loss of function medulloblastoma novel prevent public health relevance response small molecule inhibitor small molecule libraries smoothened signaling pathway tool trafficking transcription factor tumor tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): This proposal seeks to identify and characterize drug-like molecules that inhibit ciliary Hedgehog (Hh) signaling. Cells orchestrate their behaviors by communicating through secreted signals such as Hh proteins. Unlike other intercellular signals, mammalian Hh proteins are transduced through primary cilia, microtubule-based projections on the surface of many cells. The Reiter lab discovered that a central component of the Hh pathway, Smoothened (Smo), moves to the primary cilium in response to Hh stimulation where it activates the downstream pathway. The cilium is essential for Hh signaling both in embryonic development and in oncogenesis. Activating mutations in Smo cause basal cell carcinoma (BCC), the most common cancer in the U.S., and medulloblastoma, the most common solid cancer in children. A Smo antagonist was recently approved for clinical use, although cancers can become resistant quickly. Smo antagonists that act through complementary mechanisms may prevent the emergence of resistance in Hh-associated cancers. To identify novel Hh pathway antagonists and gain insight into ciliary function, the Reiter and Arkin labs completed a pilot screen for inhibitors of Smo movement to cilia. This screen identified novel inhibitors, some of which block BCC cell growth by inhibiting Smo translocation and some of which block ciliogenesis. Investigating the mechanisms by which these inhibitors act revealed previously unknown aspects of ciliary Hh signaling. This project will expand the screen, identify the molecular mechanisms underlying inhibitor action, and understand ciliary signaling. Specifically, we will: 1) screen in-house and Chemical Biology Consortium libraries, substantially expanding the chemical space that has been assessed, 2) execute secondary screens to efficiently identify inhibitors that act through previously undescribed mechanisms, 3) use newly identified antagonists, together with unique genetic tools, to uncover unrecognized steps of ciliogenesis and Smo translocation. Despite their importance to both development and disease, the mechanisms underlying ciliogenesis and Hh signaling remain unclear. The proposed investigation will provide tools to reveal how cells communicate through cilia. These compounds may also provide leads for novel chemotherapeutics, critical for preventing the emergence of resistance and relapse in Hh pathway-associated cancers.

描述（由申请人提供）：该提案旨在鉴定和表征抑制纤毛刺猬 (Hh) 信号传导的药物样分子。细胞通过 Hh 蛋白等分泌信号进行通信，与其他细胞间信号不同，哺乳动物的 Hh 蛋白是细胞间信号。 Reiter 实验室发现，Hh 通路的核心成分 Smoothened (Smo) 通过初级纤毛（许多细胞表面的微管投射）进行转导。初级纤毛响应 Hh 刺激，激活下游通路。纤毛对于胚胎发育和肿瘤发生中的 Hh 信号传导至关重要。激活 Smo 突变会导致基底细胞癌 (BCC)，这是美国最常见的癌症。髓母细胞瘤是儿童中最常见的实体癌症，最近批准用于临床，但通过补充机制发挥作用的 Smo 拮抗剂可能会迅速产生耐药性，从而防止耐药性的出现。为了识别新的 Hh 通路拮抗剂并深入了解纤毛功能，Reiter 和 Arkin 实验室完成了 Smo 纤毛运动抑制剂的初步筛选。其中一些通过抑制 Smo 易位来阻断 BCC 细胞生长，其中一些阻断纤毛发生。研究这些抑制剂的作用机制揭示了纤毛 Hh 信号传导的先前未知方面。该项目将扩大筛选范围，确定抑制剂作用的分子机制，并具体来说，我们将：1）筛选内部和化学生物学联盟库，大幅扩展已评估的化学空间，2）执行二次筛选以有效识别通过先前未描述的抑制剂发挥作用。机制，3）使用新鉴定的拮抗剂以及独特的遗传工具来揭示纤毛发生和 Smo 易位的未被识别的步骤，尽管它们对发育和疾病都很重要，但纤毛发生和 Hh 信号传导的机制仍不清楚。揭示细胞如何通过纤毛进行交流，这些化合物还可能为新型化疗药物提供线索，这对于预防 Hh 通路相关癌症的耐药性和复发至关重要。