Identification of inhibitors of hedgehog autoprocessing

刺猬自动加工抑制剂的鉴定

• 批准号: 8089846
• 负责人: CHARLES P. EMERSON
• 金额: $ 5.08万
• 依托单位: BOSTON BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8089846
• 关键词: Address; Agonist; Attention; Attenuated; Biological Assay; Biological Process; Biology; C-terminal; Cells; Chemicals; Chimeric Proteins; Cholesterol; Cholesterol Esters; Complex; Development; Drosophila genus; Embryo; Embryonic Development; Erinaceidae; Esterification; Event; Fluorescein; Fluoresceins; Fluorescence; Fluorescence Anisotropy; Fluorescence Polarization; Genes; Genetic; Genetic Transcription; Growth; Human; In Vitro; Intestines; Knowledge; Lead; Ligand Binding; Ligands; Lipids; Lung; Malignant Neoplasms; Measures; Modification; Monitor; Mutation; N-terminal; Pancreas; Pathway interactions; Pattern; Peptides; Play; Production; Prostate; Proteins; Reaction; Research; Residual state; Retina; Reverse Transcriptase Polymerase Chain Reaction; Role; Series; Signal Pathway; Signal Transduction; Signaling Molecule; Specificity; System; Therapeutic; Tumor Cell Line; Uncertainty; Variant; Western Blotting; adult stem cell; assay development; base; cancer cell; cancer stem cell; cell growth; cyclopamine; cytotoxic; cytotoxicity; high throughput screening; human SMO protein; in vitro Assay; inhibitor/antagonist; interest; meetings; mutant; public health relevance; receptor; resorufin; response; smoothened signaling pathway; stem cell division; therapeutic target; thioester; tool; transcription factor; tumor growth

DESCRIPTION (provided by applicant): Hedgehog (Hh) signaling plays an important role in embryonic patterning and adult stem cell renewal, but has recently been found also to be involved in certain stem-cell cancers, including pancreatic, small cell lung, intestinal, and prostate, all of which are notoriously difficult to treat. One of the first steps in Hh signaling is the autoprocessing of Hh protein, in which the C-terminal domain (Hh-C) catalyzes a cholesterol-dependent autocleavage reaction that leads to the production of the cholesterol ester of the N-terminal Hh domain (Hh-N), thereby yielding a signaling molecule. The secreted Hh ligand binds to the Patched receptor, leading to activation of Smoothened (Smo) and a signal transduction cascade that culminates in the activation of Gli transcription factors and the transcription of Gli and other genes. Most research on the role of Hh signaling in stem cell cancers is focused on the series of events that lie between the activation of Smo and the activation of Gli, with the aim of understanding this complex signaling pathway and in the hope to develop cancer therapeutics. In contrast, very little attention has been given to the role of Hh autoprocessing and modification, in spite of the fact that it represents the initial step in the Hh signaling pathway. Due to the absence of pharmacologic tools for its study, there is considerable uncertainty about the role of Hh autocleavage and esterification with cholesterol in Hh signaling, both in Hh signaling in the developing embryo and in the growth of Hh ligand-dependent cancers. This proposal aims to identify compounds that attenuate Hh autoprocessing as a research tool for dissecting the role of autoprocessing and the attendant cholesterol modification in Hh function, both in the many different contexts of embryonic development and in Hh ligand-dependent stem cell cancers. This approach is based on an in vitro homogeneous assay system which measures changes in fluorescence polarization that accompany the cholesterol-depended autocleavage of Hh protein. Of special interest will be the study of the effect of the inhibitors identified by this research on the growth of Hh-dependent endodermal tumor cell lines. If growth inhibition of the tumor cell lines should indeed be observed, this would suggest that Hh autoprocessing, as the first step in the Hh signaling pathway, could be a possible therapeutic target. PUBLIC HEALTH RELEVANCE: Hedgehog (Hh) signaling plays an important role in embryonic patterning and adult stem cell renewal, but has recently been found also to be responsible for certain stem-cell cancers, including pancreatic, small cell lung, intestinal, and prostate, all of which are notoriously difficult to treat. The first step in Hh signaling is an autoprocessing reaction, but there are no chemical biology tools available for studying this important step. This proposal seeks to identify inhibitors of Hh autoprocessing that can serve as tools for studying its role in normal development and in cancer and which may lead to potential cancer therapeutics.

描述（由申请人提供）：刺猬（HH）信号在胚胎模式和成人干细胞更新中起重要作用，但最近也发现与某些干细胞癌症有关，包括胰腺，小细胞肺，肠道，肠道和前列腺，众所周知，所有这些都难以治疗。 HH信号传导的第一个步骤之一是HH蛋白的自动加入，其中C末端结构域（HH-C）催化胆固醇依赖性的自启动反应，从而导致N-末端HH域（HH-N）的胆固醇酯（HH-N）的产生，从而产生信号形成。分泌的HH配体与修补的受体结合，从而导致平滑（SMO）的激活和信号转导级联反应，最终导致GLI转录因子的激活以及GLI和其他基因的转录。关于HH信号在干细胞癌中的作用的大多数研究都集中在SMO激活和GLI激活之间的一系列事件上，目的是了解这种复杂的信号传导途径，并希望开发癌症治疗剂。相比之下，尽管它代表了HH信号通路中的第一步，但很少关注HH自动处理和修改的作用。由于缺乏研究其研究的药理工具，因此HH自cleaovage和胆固醇在HH信号中的酯化作用和酯化在HH信号传导中的作用存在很大不确定性，这在发育中的胚胎中的HH信号传导以及HH配体依赖性癌症的生长中。该提案旨在鉴定减弱HH自动加入作为研究工具的化合物，以剖析自动化作用和随之而来的胆固醇修饰在HH功能中，既在胚胎发育的许多不同情况下又在HH配体依赖性干细胞癌中。这种方法基于一种体外均质测定系统，该测定系统测量伴随HH蛋白的胆固醇二甲状腺抑制性的荧光极化的变化。特别感兴趣的是该研究对HH依赖性内胚层肿瘤细胞系的生长所鉴定的抑制剂的作用的研究。如果确实应该观察到肿瘤细胞系的生长抑制作用，这将表明HH自动加入作为HH信号通路的第一步可能是可能的治疗靶标。 公共卫生相关性：刺猬（HH）信号在胚胎构图和成人干细胞更新中起重要作用，但最近也发现还负责某些干细胞癌，包括胰腺，小细胞肺，肠道，肠道和前列腺，众所周知所有这些都是难以治疗的。 HH信号传导的第一步是自动加入反应，但是没有可用于研究这一重要步骤的化学生物学工具。该提案旨在确定HH自动化的抑制剂，该抑制剂可以作为研究其在正常发育和癌症中的作用的工具，并可能导致潜在的癌症治疗剂。