Chemical Proteomic Identification of Druggable Oncogenic Transcription Factors

可药物致癌转录因子的化学蛋白质组学鉴定

• 批准号: 10357900
• 负责人: Liron Bar-Peled
• 金额: $ 19.24万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2024-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10357900
• 关键词: Address; Adopted; Antineoplastic Agents; Applications Grants; Binding Proteins; Cancer cell line; Chemicals; Chromatin; Clinical; Complex; Credentialing; Cysteine; Detection; Development; Disease Progression; Enzymes; Genes; Genetic Transcription; Goals; Growth; Hour; Human Genetics; Immunoprecipitation; Label; Libraries; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Metabolic; Methods; Mutate; Oncogenes; Oncogenic; Organelles; Pharmaceutical Preparations; Pharmacology; Phosphotransferases; Physiological; Play; Protein Family; Proteins; Proteome; Proteomics; Research; Resolution; Role; Sampling; Site; Specificity; Speed; Technology; Validation; Work; anti-cancer therapeutic; anticancer research; base; cancer cell; cancer therapy; chemical property; chemoproteomics; convict; detection sensitivity; drug development; drug discovery; established cell line; experimental study; functional genomics; inhibitor; insight; next generation; protein function; protein protein interaction; small molecule; small molecule inhibitor; stable cell line; stem; success; targeted treatment; tool; transcription factor; tumorigenesis

Project Summary Advances in human genetics have identified +400 genes that when amplified or mutated promote tumorigenesis. While there has been huge success in developing drugs for kinases and metabolic enzymes deregulated in cancer, they represent only a small fraction of the cancer drivers discovered to date. A major challenge in cancer research is developing drugs for oncogenic drivers. The vast majority (~80%) of these cancer drivers remain undrugged, including one of the largest classes, transcription factors (TFs) which account for ~19% of oncogenes. These TFs are normally required during development, however, are hijacked during tumorigenesis providing malignant cells with the plasticity required for unchecked proliferation. Although these oncogenic TFs have been biologically credentialed, they are historically considered difficult to target with small-molecule inhibitors, limiting the development of transformative cancer therapies. To overcome these challenges in cancer drug discovery and address the clear unmet needs in cancer treatment, our lab adopts state-of-the-art chemical proteomic platforms that both radically expand the druggable protein landscape in cancer and allow us to pinpoint which proteins are amenable to small molecule inhibition. These chemical proteomic technologies combine the specificity of chemical probes which only react with proteinaceous cysteines with the comprehensive analytical scale of next generation proteomics. Cysteines play critical roles in protein function and are the targets of multiple clinically approved inhibitors. By profiling their interaction with covalent drug-like fragments, we recently discovered that a much larger extant of the proteome than originally predicted is amenable to covalent druggability. While these chemical proteomic approaches have transformed our notion of which proteins are druggable, they remain ill-equipped (due to sensitivity of detection) to determine cysteine druggability on low abundance oncogenic transcription factors. In this grant application, we build on our core chemical proteomic platform and incorporate advances in protein and organelle enrichment technologies to prosecute the druggabilty of high-priority oncogenic TFs by developing two conceptually new and complimentary approaches: 1) By isolating chromatin bound proteins, we enrich for active TFs, enabling us to use traditional chemical proteomic approaches to provide a high content map of druggable cysteines in oncogenic TFs. 2) We develop Enrichment Cysteine Druggability Mapping (ECDM) which allows us to systematically immunoprecipitate and enrich low-abundance TFs and rapidly interrogate the druggabilty of cysteines found in these factors in a period of 18 minutes compared to 30 hours using standard approaches. The research proposed herein, takes full advantage of advances in human genetics and functional genomics and combines them with ultra-high throughput chemical proteomic technologies to define the druggability of TF cancer drivers, a critical first step in the development of targeted therapies. If successful, the development of these drug-discovery platforms has the potential to reshape the next-generation of targeted anti-cancer therapeutics.

项目摘要 人类遗传学的进步已经鉴定出+400个基因，当扩增或突变促进肿瘤发生时。 尽管在开发激酶和代谢酶的药物方面取得了巨大成功，但在 癌症，它们仅占迄今为止发现的癌症驱动因素的一小部分。癌症的主要挑战 研究正在开发致癌驱动因素的药物。这些癌症司机中绝大多数（约80％）仍然存在 未经策划的人，包括最大的班级之一，转录因子（TF），占约19％的19％ 癌基因。但是，在发育过程中通常需要这些TF，但是在肿瘤发生期间被劫持 为恶性细胞提供未检查的增殖所需的可塑性。尽管这些致癌TF 在生物学上具有认证，历史上被认为很难用小​​分子靶向 抑制剂，限制了变革性癌症疗法的发展。克服这些癌症的挑战 药物发现并满足癌症治疗中明确未满足的需求，我们的实验室采用了最先进的化学 蛋白质组学平台既从根本上扩展了癌症中的可毒蛋白景观，又可以确定 哪种蛋白质可与小分子抑制作用。这些化学蛋白质组学技术结合了 化学探针的特异性仅与蛋白质半胱氨酸反应 下一代蛋白质组学的比例。半胱氨酸在蛋白质功能中起关键作用，是多重目标 临床认可的抑制剂。通过分析它们与共价药物片段的相互作用，我们最近 发现蛋白质组的现存比最初预测的要大得多 可药物。尽管这些化学蛋白质组学方法改变​​了我们的观念 可吸毒，它们仍然装备不足（由于检测的敏感性），以确定低于低的半胱氨酸吸毒性 丰度致癌转录因子。在此赠款应用程序中，我们以核心化学蛋白质组学为基础 平台并纳入蛋白质和细胞器富集技术的进步，以起诉 通过开发两种概念上的新方法和免费的方法来实现高优先性致癌TF的药物： 1）通过隔离染色质结合的蛋白，我们丰富了活性TF，使我们能够使用传统的化学物质 蛋白质组学的方法可提供高含量毒素的高含量含量的含量TFS的含量图。 2）我们发展 富集半胱氨酸的可药用映射（ECDM），使我们能够系统地免疫沉淀和 富集低丰度的TF，并迅速询问在这些因素中发现的半胱氨酸的药物的药物。 使用标准方法为18分钟，而30小时。本文提出的研究完整 人类遗传学和功能基因组学的进步优势，并将它们与超高相结合 吞吐量化学蛋白质组学技术来定义TF癌驱动因素的可药物，这是关键的第一步 靶向疗法的发展。如果成功，这些药物发现平台的开发具有 重塑目标抗癌治疗剂的下一代的潜力。