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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要人类遗传学的进展已鉴定出超过 400 个基因，这些基因在扩增或突变时会促进肿瘤发生。尽管在开发用于解除对激酶和代谢酶的管制的药物方面取得了巨大成功癌症，它们仅代表迄今为止发现的癌症驱动因素的一小部分。癌症领域的重大挑战研究正在开发用于致癌驱动因素的药物。这些癌症驱动因素中的绝大多数（~80%）仍然存在未用药，包括最大的一类转录因子 (TF)，约占 19% 癌基因。这些转录因子通常在发育过程中需要，但在肿瘤发生过程中被劫持为恶性细胞提供不受抑制的增殖所需的可塑性。尽管这些致癌 TF 已获得生物学认证，历史上认为它们难以用小分子靶向抑制剂，限制了转化性癌症疗法的发展。为了克服癌症方面的这些挑战药物发现并解决癌症治疗中明显未满足的需求，我们的实验室采用最先进的化学品蛋白质组学平台既可以从根本上扩展癌症中的可药物蛋白质景观，又可以让我们精确定位哪些蛋白质易于受到小分子抑制。这些化学蛋白质组技术结合了化学探针的特异性，仅与蛋白质半胱氨酸反应，具有全面的分析能力下一代蛋白质组学的规模。半胱氨酸在蛋白质功能中发挥着关键作用，并且是多种蛋白质的靶标。临床批准的抑制剂。通过分析它们与共价药物样片段的相互作用，我们最近发现比最初预测的更大的现存蛋白质组适合共价成药性。虽然这些化学蛋白质组学方法已经改变了我们对哪些蛋白质是蛋白质的概念尽管可成药，但他们仍然没有能力（由于检测的敏感性）来确定低半胱氨酸的成药性丰富的致癌转录因子。在这项拨款申请中，我们以我们的核心化学蛋白质组学为基础平台并结合蛋白质和细胞器富集技术的进步来起诉通过开发两种概念上新的互补方法来提高高优先级致癌转录因子的成药性： 1) 通过分离染色质结合蛋白，我们富集了活性转录因子，使我们能够使用传统的化学物质蛋白质组学方法提供致癌 TF 中可药物半胱氨酸的高含量图谱。 2）我们开发富集半胱氨酸成药性图谱 (ECDM)，使我们能够系统地免疫沉淀和富集低丰度 TF，并快速询问一段时间内这些因子中发现的半胱氨酸的成药性使用标准方法需要 30 小时，而缩短了 18 分钟。本文提出的研究充分考虑了利用人类遗传学和功能基因组学的进步并将其与超高通过化学蛋白质组学技术来定义 TF 癌症驱动因素的成药性，这是关键的第一步靶向治疗的发展。如果成功，这些药物发现平台的开发将具有以下优势：重塑下一代靶向抗癌疗法的潜力。