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.

研究摘要 人类基因组编码1,600多个转录因子（TFS），以及其他 共同执行调节的辅因子，染色质调节剂和结构蛋白 指示在核DNA中编码。这些蛋白质的功能障碍，统称 由于基因调节蛋白（GRP），已知会驱动多种疾病，例如癌症， 在癌症中，这些蛋白质经常是 重新排列并融合以创建新蛋白质，从而导致主动性和进展 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，但 由于在体外对其活性建模时，GRP被认为是不受限制的。 我们通过实施一种内部功能蛋白质组学药物解决了这些缺点 量化小分子对GRP抽象的影响的发现平台 与基因组结合在各种细胞和组织类型中。该平台基于染色质 通过盐分离提取，与数据独立分析质谱法耦合 （国际象棋），最近报道。在此提案中，我们将将这项技术应用于 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）重新排列的白血病，以执行致癌融合蛋白的技术开发 蛋白质组学策略。在AML和所有患者的子集中发现了MLL重排， 通常在儿童中，并且仍然受到挑战，以治疗现有的治疗选择。一些 目前正在临床试验中 用于验证国际象棋DIA分析的准确性以报告靶向MLL的能力 化合物破坏活细胞中的MLL复合物。带有经过验证的MLL棋子测定法， 然后，我们将使用试点屏幕，以使用筛选设置证明评估的实用性 国家癌症研究所的机械多样性化合物集，并补充已知的 MLL复合物的抑制剂。这些化合物含有一系列生物活性阵列，许多 其中通过未知机制起作用。这提供了找到新的机会 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 为了使MLL开发许多常见的致癌融合的国际象棋分析，然后 开发一种将这些测定统一的方法中的单个，统一的，细胞内测定的致癌方法 融合蛋白。