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

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）：在癌症中变得过度活跃的转录因子是有希望但未经测试的治疗靶点。 c-Myc（一种主调节转录因子）的激活是人类恶性肿瘤中最常见的致癌事件之一。 c-Myc 失活似乎会引发癌基因成瘾和肿瘤消退。此外，c-Myc 是唯一已在体内证实真正致癌基因成瘾的致癌基因之一。使用小分子灭活 c-Myc 可能提供一种通用且有效的癌症治疗方法，因为 Myc 是最常见的恶性肿瘤驱动因素之一。该提案的目标是发现、表征和优化可用作 Myc 依赖性转录探针的 c-Myc 直接小分子调节剂，并阐明 c-Myc 作为治疗靶点的潜力。尽管 c-Myc 参与多种蛋白质-蛋白质相互作用，但开发抑制剂的工作重点是调节 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 异二聚体发挥作用，并且具有 Max 独立功能的新型探针可能有助于表征它们在未转化细胞和恶性细胞中的相关性。为了鉴定 c-Myc 功能的新型探针，我们进行了一项高通量结合测定，涉及用全长 c-Myc 探测的小分子微阵列。在报告基因测定中，鉴定了几种直接结合物并发现其影响 Myc 依赖性转录。在这里，我们建议使用这些化合物作为开发 c-Myc 功能的新型小分子探针的起点，遵循化学生物学界所接受的开发高质量探针的原则。本文概述的研究旨在建立源自我们之前研究的直接 c-Myc 调节剂的作用机制，并优化探针以研究未转化细胞和恶性细胞中的 Myc 功能。因此，我们的具体目标是： 目标 1) 通过在一系列无细胞和基于细胞的检测中评估候选探针来表征其作用机制，包括将化合物区分为两个探针类别的研究 - Myc/Max 调节剂和最大独立探针。目标 2) 研究任一类别中选定的 c-Myc 候选探针的结构-活性关系，并对其进行优化以提高效力、选择性和理化性质。目标 3) 利用优化且机制不同的 c-Myc 探针来探究 c-Myc 的 Max 依赖性和 Max 独立功能在细胞增殖、转录和肿瘤发生中的作用。鉴于几乎所有病例都涉及 MYC 基因的平衡易位和癌蛋白的过度表达，伯基特淋巴瘤将作为这些机制研究的主要模型系统。通过这些累积的努力，我们希望构建一套新的高质量 Myc 介导的转录探针，可用于阐明 c-Myc 作为直接治疗靶点的作用，并作为涉及异常恶性肿瘤的转化研究的起点。 Myc 函数。