Developing Direct Small-Molecule Probes of Myc-Dependent Transcription

开发 Myc 依赖性转录的直接小分子探针

• 批准号: 8826461
• 负责人: Angela Nicole Koehler
• 金额: $ 34.81万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-30 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8826461
• 关键词: Affect; Affinity; Binding; Biological Assay; Biological Models; Biology; Burkitt Lymphoma; Cancer Biology; Cell Cycle; Cell Cycle Regulation; Cell Line; Cell Proliferation; Cells; Chemicals; Collection; Communities; Community Developments; DNA; DNA Binding; DNA Damage; Dependency; Equilibrium; Event; Gene Expression; Genes; Genetic Transcription; Goals; Growth; Half-Life; Human; In Vitro; Institutes; Knowledge; Lead; Length; Literature; MYC gene; Malignant Neoplasms; Mediating; Normal Cell; Oncogene Proteins; Oncogenes; Oncogenic; Peptides; Play; Post-Translational Protein Processing; Property; Proteins; Proteomics; Proto-Oncogene Proteins c-myc; Regenerative Medicine; Relative (related person); Reporter Genes; Research; Role; Series; Structure-Activity Relationship; Synthesis Chemistry; analog; base; c-myc Genes; cancer cell; cancer therapy; cellular targeting; design; experience; improved; in vivo; inhibitor/antagonist; neoplastic cell; novel; oncogene addiction; overexpression; prevent; programs; protein protein interaction; research study; small molecule; stem; therapeutic target; tool; transcription factor; translational study; tumor; tumorigenesis; Affect; Affinity; Binding; Biological Assay; Biological Models; Biology; Burkitt Lymphoma; Cancer Biology; Cell Cycle; Cell Cycle Regulation; Cell Line; Cell Proliferation; Cells; Chemicals; Collection; Communities; Community Developments; DNA; DNA Binding; DNA Damage; Dependency; Equilibrium; Event; Gene Expression; Genes; Genetic Transcription; Goals; Growth; Half-Life; Human; In Vitro; Institutes; Knowledge; Lead; Length; Literature; MYC gene; Malignant Neoplasms; Mediating; Normal Cell; Oncogene Proteins; Oncogenes; Oncogenic; Peptides; Play; Post-Translational Protein Processing; Property; Proteins; Proteomics; Proto-Oncogene Proteins c-myc; Regenerative Medicine; Relative (related person); Reporter Genes; Research; Role; Series; Structure-Activity Relationship; Synthesis Chemistry; analog; base; c-myc Genes; cancer cell; cancer therapy; cellular targeting; design; experience; improved; in vivo; inhibitor/antagonist; neoplastic cell; novel; oncogene addiction; overexpression; prevent; programs; protein protein interaction; research study; small molecule; stem; therapeutic target; tool; transcription factor; translational study; tumor; tumorigenesis

DESCRIPTION (provided by applicant): Transcription factors that become overactive in cancers are promising yet untested targets for therapeutics. Activation of c-Myc, a master regulator transcription factor, is one of the most common oncogenic events in human malignancies. Inactivation of c-Myc appears to elicit oncogene addiction and tumor regression. Furthermore, c-Myc is one of the only oncogenes for which true oncogene addiction has been demonstrated in vivo. Inactivation of c-Myc using small molecules may provide a general and effective approach to treating cancers as Myc is one of the most common drivers of malignancy. The goal of this proposal is to discover, characterize, and optimize direct small-molecule modulators of c-Myc that may be used as probes of Myc- dependent transcription and to clarify the potential of c-Myc as a therapeutic target. Although c-Myc participates in several protein-protein interactions, efforts to develop inhibitors have focused on modulating the interaction between c-Myc and heterodimer partner Max. Recent evidence indicates c-Myc can also function independently of the Myc/Max heterodimer and novel probes of Max-independent functions may help characterize their relevance in both untransformed and malignant cells. In an effort to identify novel probes of c-Myc function, we executed a high-throughput binding assay involving small-molecule microarrays probed with full-length c-Myc. Several direct binders were identified and found to affect Myc-dependent transcription in a reporter gene assay. Here we propose to use these compounds as starting points for developing novel small- molecule probes of c-Myc function, adhering to principles accepted by the chemical biology community for the development of high-quality probes. The studies outlined herein are aimed at establishing mechanism of action for direct c-Myc modulators stemming from our previous studies and optimizing the probes to enable studies of Myc functions in untransformed and malignant cells. Accordingly, our specific aims are: Aim 1) Characterize the mechanism of action for the probe candidates by evaluating them in a series of cell-free and cell-based assays, including studies to differentiate compounds into two probe classes- Myc/Max modulators and Max- independent probes. Aim 2) Investigate structure-activity relationships for selected candidate probes of c-Myc, from either class, and optimize them to improve potency, selectivity, and physicochemical properties. Aim 3) Leverage optimized and mechanistically distinct probes of c-Myc to interrogate the roles of Max-dependent and Max-independent functions of c-Myc in cell proliferation, transcription, and tumorigenesis. Burkitt's lymphoma will serve as the primary model system for these mechanistic studies given the fact that nearly all cases involve balanced translocation of the MYC gene and overexpression of the oncoprotein. Through these cumulative efforts we hope to build a novel set of high-quality probes of Myc-mediated transcription that may be used to clarify the role of c-Myc as a direct therapeutic target and serve as starting points for translational studies involving malignancies with aberrant Myc function.

描述（由申请人提供）：在癌症中过度活跃的转录因子是有希望但未经测试的治疗靶标。 C-MYC的激活是一种主调节转录因子，是人类恶性肿瘤中最常见的致癌事件之一。 C-MYC的灭活似乎会引起癌基因成瘾和肿瘤消退。此外，C-MYC是唯一在体内证明了真正的癌基因成瘾的癌基因之一。使用小分子灭活C-MYC可能会提供一种普遍有效的方法来治疗癌症，因为MYC是最常见的恶性驱动因素之一。该提案的目的是发现，表征和优化C-MYC的直接小分子调节剂，该调节剂可以用作依赖Myc依赖的转录探针，并阐明C-MYC作为治疗靶标的潜力。尽管C-MYC参与了几种蛋白质 - 蛋白质相互作用，但开发抑制剂的努力致力于调节C-MYC和异二聚体伴侣Max之间的相互作用。最近的证据表明，C-MYC也可以独立于MYC/MAX HETRODIMER起作用，而新颖的非依赖性功能的新探针可能有助于表征它们在未转换和恶性细胞中的相关性。为了确定C-MYC功能的新型探针，我们执行了一个高通量结合测定法，该测定法涉及使用全长C-MYC探测的小分子微阵列。鉴定出几种直接粘合剂并发现在报告基因测定中影响MYC依赖性转录。在这里，我们建议将这些化合物用作开发新型小分子探针C-MYC功能的起点，并遵守化学生物学界接受的原理来开发高质量的探针。本文概述的研究旨在建立源于我们先前的研究并优化探针的直接C-MYC调节剂的作用机理，以使MYC功能在未转换和恶性细胞中进行研究。因此，我们的具体目的是：目标1）通过在一系列无细胞和基于细胞的测定中评估探针候选者的作用机理，包括将化合物分为两个探针类的研究 - MYC/MAX Max调节剂和最大独立探针。 AIM 2）研究C-Myc选定的候选探针（从任何一个类别）中研究结构 - 活性关系，并优化它们以提高效力，选择性和理化特性。目标3）利用了C-MYC的优化和机械上不同的探针来询问C-Myc在细胞增殖，转录和肿瘤发生中的最大依赖性和最大依赖性功能的作用。伯基特的淋巴瘤将作为这些机械研究的主要模型系统，因为几乎所有病例都涉及MYC基因的平衡易位和癌蛋白的过表达。通过这些累积努力，我们希望建立一组新型的MYC介导的转录的高质量探针，这些探针可用于阐明C-Myc作为直接治疗靶点的作用，并作为涉及涉及异常MYC功能的恶性肿瘤的转化研究的起点。