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.

研究概要 人类基因组编码超过 1,600 个转录因子 (TF)，以及其他 共同执行调节的辅因子、染色质调节因子和结构蛋白 这些蛋白质的功能障碍，统称为核 DNA 内编码的指令。 作为基因调节蛋白 (GRP)，已知会导致多种疾病，例如癌症、 在癌症中，这些蛋白质经常与炎症相关和神经系统疾病相关。 重新排列和融合以产生新的蛋白质，从而导致启动和进展 尽管这些蛋白质很重要，但各种类型的白血病、肉瘤和其他肿瘤。 由于体外活性建模存在挑战，GRP 被认为是不可成药的。 我们通过实施细胞内功能蛋白质组药物解决了这些缺点 量化小分子对 GRP 丰度影响的发现平台 该平台基于染色质，与多种细胞和组织类型的基因组结合。 通过盐分离提取，结合数据独立分析质谱法 （ChESS-DIA），在最近报道的这项提案中，我们将应用这项技术。 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 蛋白质组学策略在一部分 AML 和 ALL 患者中发现了 MLL 重排， 常见于儿童，并且用现有的治疗方案来治疗仍然具有挑战性。 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 测定报告 MLL 靶向能力的准确性 通过经过验证的 MLL ChESS-DIA 检测，可以破坏活细胞中的 MLL 复合物。 然后，我们将进行试点筛选，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制多样性复合物集补充了已知的 MLL 复合物的抑制剂这些化合物含有多种生物活性。 它通过未知的机制起作用，这提供了寻找新的机会。 最后，我们将使用开发的路线图。 MLL 为许多常见的致癌融合开发 ChESS-DIA 检测，然后 开发一种方法将这些测定统一到一个单一的、统一的细胞内致癌测定中 融合蛋白。