Identification and characterization of chemical probes for interrogation of the NEK family of kinases in cancer

用于研究癌症中 NEK 激酶家族的化学探针的鉴定和表征

• 批准号: 10503430
• 负责人: Matthew E. Burow
• 金额: $ 63.61万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10503430
• 关键词: 3-Dimensional; Active Sites; Address; Antineoplastic Agents; Binding; Biological; Biological Assay; Biological Process; Biology; Cell Cycle; Cell Cycle Regulation; Cell Line; Cells; Cellular biology; Chemicals; Communities; DNA Damage; DNA biosynthesis; Data; Development; Diabetes Mellitus; Disadvantaged; Disease; Disease Progression; Drug Targeting; Environment; Etiology; FDA approved; Family; Family member; Foundations; Free Will; Funding; Generations; Genome; Health; Human; Individual; Inflammation; Inflammatory Bowel Diseases; Knock-out; Knowledge; Left; Link; Malignant Neoplasms; Measures; Medicine; Mission; Mitosis; Modality; Molecular; Nature; Neurons; Oncology; Output; Pathologic; Pathway interactions; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phosphotransferases; Physiological; Play; Process; Protein Family; Protein Kinase; Proteins; Proteome; RNA Splicing; Reagent; Reporter; Research; Research Personnel; Resources; Role; Scientist; Signal Pathway; Signal Transduction; System; Therapeutic; United States National Institutes of Health; Validation; Work; base; cancer therapy; cell motility; chronic inflammatory disease; ciliopathy; cilium biogenesis; design; drug development; drug discovery; drug quality; druggable target; holistic approach; inhibitor; insight; knock-down; lead optimization; member; migration; new therapeutic target; programs; response; scaffold; small molecule; success; tool

ABSTRACT / PROJECT SUMMARY Kinases, a class of proteins with more than 500 members in the human proteome, are important regulators of biological processes in health and disease. Kinases have proven to be excellent drug targets with more than 70 FDA approved medicines that target kinases. Despite this success, most kinases are understudied, and details of their functions are poorly understood. The NEK family of 11 kinases (NEK1 through NEK11) is a particularly understudied set of kinases that play roles in key biological processes like the cell cycle, ciliogenesis, and the DNA damage response (DDR), all with relevance to cancer and human health. These kinases have emerging links to numerous cancers, diabetes, inflammatory bowel disease, ciliopathies, and ALS. In this project we will use an efficient kinase systems-based approach to create an enabling suite of chemical probes, assays, reagents, and molecular tools to identify NEK family members that have key roles in cancer. These high-quality compounds and reagents we generate, which we will freely share, will allow scientists to build a deep understanding of the physiological and pathological roles members of the NEK family play. In Aim 1 we will create potent and selective inhibitors of each NEK using iterative medicinal chemistry and state of the art in cell target engagement assays. In a complementary effort for this aim we will also create inducible NEK knockdown cell lines. In Aim 2, using compounds and the NEK knockdown lines, we will evaluate the role and importance of each NEK in a suite of NEK and oncology-relevant cell health and cell biology signaling assays, measuring effects on proliferation, migration, the cell-cycle, DNA replication, and ciliogenesis. In Aim 3 we will experimentally determine the substrates of each NEK, locate the NEKs in broader kinase-dependent signaling pathways, and develop genetically targetable kinase activity reporters for tracking NEK activity within the endogenous cellular environment. Output from this project will include potent and selective NEK inhibitors, NEK family-wide assays, details on the impact of NEK inhibition and knockdown on key cancer processes, molecular tools, and NEK substrate and pathway information. Successful completion will provide a framework and the resources needed to validate individual NEKs as high quality, druggable targets for the treatment of cancer.

摘要/项目摘要 蛋白激酶是人类蛋白质组中一类有500多个成员的蛋白质，是重要的调节因子。 健康和疾病中的生物过程。激酶已被证明是极好的药物靶点，有70多种 FDA批准了针对激酶的药物。尽管取得了这样的成功，但大多数激酶还没有得到充分的研究，而且细节 对它们的功能知之甚少。NEK家族由11个蛋白组成(NEK1到NEK11)，是一种特殊的 未被充分研究的一组在细胞周期、纤毛发生和细胞周期等关键生物学过程中发挥作用的激酶 DNA损伤反应(DDR)，都与癌症和人类健康有关。这些激酶已经出现了 与多种癌症、糖尿病、炎症性肠病、纤毛病和肌萎缩侧索硬化症有关。在这个项目中，我们将 使用一种高效的基于激酶系统的方法来创建一套使化学探针、分析、 试剂和分子工具，以确定在癌症中起关键作用的NEK家族成员。这些高品质的 我们产生的化合物和试剂，我们将免费分享，将允许科学家建立一个深层次的 了解NEK家族成员所扮演的生理和病理角色。在目标1中，我们将 使用迭代药物化学和细胞内最新技术创建每个NEK的有效和选择性抑制剂 目标参与度分析。为了实现这一目标，我们还将创造可诱导的NEK击倒 细胞系。在目标2中，使用化合物和NEK击倒线，我们将评估 一套NEK和肿瘤学相关的细胞健康和细胞生物学信号分析中的每个NEK，测量 对增殖、迁移、细胞周期、DNA复制和纤毛发生的影响。在《目标3》中，我们将 实验确定每个NEK的底物，在更广泛的激酶依赖信号中定位NEK 途径，并开发基因靶向的激酶活性报告器，用于跟踪NEK在 内源性细胞环境。该项目的产出将包括有效和选择性的NEK抑制剂，NEK 家庭范围的分析，关于NEK抑制和敲除对关键癌症过程的影响的细节，分子 工具，以及NEK底物和途径信息。成功完成后，将提供一个框架和 验证个别NeKs作为高质量、可用药的癌症治疗靶点所需的资源。