Simultaneous pharmacological profiling of oncogenic gene fusion proteins in cancer

癌症中致癌基因融合蛋白的同时药理学分析

• 批准号: 10378326
• 负责人: Alexander Federation
• 金额: $ 45.5万
• 依托单位: TALUS BIOSCIENCE, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10378326
• 关键词: Address; Binding Proteins; Biochemical; Biological Assay; Calibration; Cell Nucleus; Cells; Cellular Assay; Child; Chimeric Proteins; Chromatin; Clinical Trials; Collaborations; Complex; Coupled; Couples; DNA; DNA Binding; Data; Development; Digestion; Disease; Drug Targeting; Environment; Functional disorder; Fusion Oncogene Proteins; Gene Expression; Gene Fusion; Genetic; Genetic Transcription; Genome; Goals; Human Genome; Inflammation; Institutes; Instruction; Label; Length; Leukemic Cell; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Measures; Methods; Mixed-Lineage Leukemia; Modeling; Multiprotein Complexes; National Cancer Institute; Neurologic; Nuclear; Oncogenes; Oncogenic; Organelles; Patients; Peptides; Performance; Pharmaceutical Preparations; Pharmacology; Phase; Proteins; Proteome; Proteomics; Regulator Genes; Reporting; Research; Science; Small Business Innovation Research Grant; Sodium Chloride; Specificity; Stimulus; Structural Protein; System; Talus; Technology; Testing; Therapeutic; Tissues; Universities; Validation; Variant; Washington; base; cancer cell; cofactor; data quality; drug candidate; drug development; drug discovery; experimental study; genetic regulatory protein; in vitro activity; inhibitor; leukemia; member; prevent; quality assurance; response; sarcoma; screening; small molecule; small molecule inhibitor; technology development; tool; transcription factor; tumor

RESEARCH SUMMARY The human genome encodes more than 1,600 transcription factors (TFs), along with additional cofactors, chromatin regulators, and structural proteins that collectively execute the regulatory instructions encoded within the nuclear DNA. Dysfunctions of these proteins, collectively known as Gene Regulatory Proteins (GRPs), are known to drive multiple diseases such as cancer, inflammation-related, and neurological conditions. In cancer, these proteins are frequently rearranged and fused to create new proteins which cause the initiation and progression of various types of leukemia, sarcoma and other tumors. Despite the importance of these proteins, GRPs have been considered undruggable due to challenges in modeling their activity in vitro. We have solved these shortcomings by implementing an in-cell functional proteomics drug discovery platform that quantifies the effects of small-molecules on the abundance of GRPs bound to the genome in a diversity of cell and tissue types. The platform is based on Chromatin Extraction by Salt Separation, coupled to Data Independent Analysis mass spectrometry (ChESS-DIA), which was recently reported. In this proposal, we will apply this technology for drug development of oncogenic fusion proteins. First, we will use Mixed Lineage Leukemia (MLL) rearranged leukemia to perform technology development of the oncogenic fusion protein proteomics strategy. MLL rearrangements are found in a subset of AML and ALL patients, commonly in children, and remain challenging to treat with existing therapeutic options. Several drug candidates for MLL-rearranged leukemia are currently in clinical trials, and these will be used to validate the accuracy of the ChESS-DIA assay for reporting the ability of MLL-targeting compounds to disrupt the MLL complex in live cells. With a validated MLL ChESS-DIA assay, we will then conduct a pilot screen to prove the assay’s utility in a screening setting, using the National Cancer Institute’s Mechanistic Diversity compound set supplemented with known inhibitors of the MLL complex. These compounds contain a diverse array of bioactivities, many of which act through unknown mechanisms. This provides an opportunity to find new compounds capable of disrupting the MLL complex. Lastly, we will use the roadmap developed for MLL to develop ChESS-DIA assays for many of the common oncogenic fusions, then develop a method to unify these assays together in a single, unified, in-cell assay for oncogenic fusion proteins.

研究综述 人类基因组编码超过1，600种转录因子（TF），沿着有额外的转录因子。 辅因子、染色质调节因子和共同执行调节的结构蛋白质。 编码在核DNA中的指令。这些蛋白质的功能障碍， 作为基因调节蛋白（GRP），已知驱动多种疾病，例如癌症， 炎症相关和神经系统疾病。在癌症中，这些蛋白质通常 重组和融合，创造新的蛋白质，引起的起始和进展， 各种类型的白血病、肉瘤和其他肿瘤。尽管这些蛋白质很重要， GRP由于在体外模拟其活性方面的挑战而被认为是不可药用的。 我们通过实施细胞内功能蛋白质组学药物解决了这些缺点 发现平台，量化小分子对GRP丰度的影响 与多种细胞和组织类型的基因组结合。该平台基于染色质 通过盐分离提取，与数据独立分析质谱联用 （ChESS-DIA），最近报道。在本提案中，我们将把这项技术应用于 致癌融合蛋白的药物开发。首先，我们将使用混合谱系白血病 (MLL)重排白血病进行致癌融合蛋白的技术开发 蛋白质组学策略在AML和ALL患者的亚组中发现MLL重排， 通常在儿童中，并且仍然具有挑战性，难以用现有的治疗方案进行治疗。几 治疗MLL重排白血病的候选药物目前正在进行临床试验，这些药物将在 用于验证ChESS-DIA检测报告MLL靶向能力的准确性 化合物以破坏活细胞中的MLL复合物。使用经验证的MLL ChESS-DIA测定， 然后，我们将进行一个试点筛选，以证明该测定在筛选环境中的实用性，使用 国家癌症研究所的机械多样性化合物集补充了已知的 MLL复合物的抑制剂。这些化合物具有多种生物活性， 其中通过未知的机制发挥作用。这提供了一个机会， 能够破坏MLL复合物的化合物。最后，我们将使用开发的路线图 对于MLL开发许多常见致癌融合的ChESS-DIA检测，然后 开发一种方法，将这些检测方法统一在一个单一的、统一的细胞内致癌检测方法中， 融合蛋白