Identification and characterization of chemical probes for interrogation of the NEK family of kinases in cancer (Diversity Supplement - Belgodere)

用于询问癌症中 NEK 激酶家族的化学探针的鉴定和表征（多样性补充 - Belgodere）

基本信息

批准号：
10745843
负责人：
Matthew E. Burow
金额：
$ 11.02万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
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

项目摘要

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/项目摘要激酶是人类蛋白质组中一类拥有 500 多个成员的蛋白质，是重要的调节因子健康和疾病的生物过程。激酶已被证明是极好的药物靶标，有 70 多种 FDA 批准了针对激酶的药物。尽管取得了这一成功，但大多数激酶尚未得到充分研究，详细信息对它们的功能知之甚少。 NEK 家族由 11 种激酶（NEK1 至 NEK11）组成，是一个特别重要的激酶家族。一组尚未被研究的激酶，它们在细胞周期、纤毛发生和细胞周期等关键生物过程中发挥作用 DNA 损伤反应 (DDR)，所有这些都与癌症和人类健康相关。这些激酶已经出现与多种癌症、糖尿病、炎症性肠病、纤毛病和肌萎缩侧索硬化症（ALS）有关。在这个项目中我们将使用基于激酶系统的有效方法来创建一套化学探针、分析、用于识别在癌症中起关键作用的 NEK 家族成员的试剂和分子工具。这些高品质我们将自由分享我们产生的化合物和试剂，这将使科学家能够建立深入的研究了解 NEK 家族成员所发挥的生理和病理作用。在目标 1 中，我们将使用迭代药物化学和最先进的细胞技术创建每个 NEK 的有效和选择性抑制剂目标参与分析。为了实现这一目标，我们还将创造诱导性 NEK 击倒细胞系。在目标 2 中，使用化合物和 NEK 敲除系，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，我们将通过实验确定每个 NEK 的底物，在更广泛的激酶依赖性信号传导中定位 NEK 途径，并开发遗传靶向激酶活性报告基因，用于跟踪 NEK 内的活性内源性细胞环境。该项目的产出将包括有效且选择性的 NEK 抑制剂、NEK 全家族检测、NEK 抑制和敲低对关键癌症过程的影响的详细信息、分子工具，以及 NEK 底物和途径信息。成功完成将提供一个框架和验证单个 NEK 作为癌症治疗的高质量、可药物靶标所需的资源。