Targeting allosteric scaffolding functions of Aurora kinase A in cancer

靶向癌症中极光激酶 A 的变构支架功能

• 批准号: 10373096
• 负责人: Nicholas Mark Levinson
• 金额: $ 34.75万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10373096
• 关键词: Active Sites; Affect; Affinity; Agreement; Animal Model; Area; Binding; Binding Proteins; Binding Sites; Cancer cell line; Cell Death; Cell Line; Cells; Centrosome; Clinical; Clinical Trials; Complex; Computer Models; Crystallization; Data; Development; Disease; Electrons; Elements; Eukaryotic Cell; Exhibits; Family; Fluorescence Resonance Energy Transfer; Goals; Human; In Vitro; Infant; Lead; Libraries; MYC Family Protein; MYCN gene; Magnetic Resonance Spectroscopy; Malignant Neoplasms; Malignant neoplasm of liver; Measurement; Measures; Mitotic; Modality; Molecular; Molecular Conformation; Motion; N-Myc Protein; N-terminal; Neuroblastoma; Neuroendocrine Prostate Cancer; Oncogenes; Oncogenic; Oncoproteins; Paper; Patients; Phosphorylation; Phosphotransferases; Positioning Attribute; Prognosis; Protein Kinase; Proteins; Proto-Oncogene Proteins c-myc; Publishing; Regulatory Element; Resolution; Roentgen Rays; SKP Cullin F-Box Protein Ligases; Series; Shapes; Signal Transduction; Solid Neoplasm; Spectrum Analysis; Structure; Testing; Therapeutic; Time; Ubiquitination; Work; X-Ray Crystallography; aurora kinase A; base; c-myc Genes; cancer cell; cancer therapy; cell growth; clinical candidate; drug development; experimental study; high risk; inhibitor; insight; kinase inhibitor; molecular modeling; multicatalytic endopeptidase complex; nanosecond; neuroblastoma cell; next generation; overexpression; prevent; prostate cancer cell line; response; scaffold; sensor; small molecule inhibitor; success; therapeutic target; tool; transcription factor; tumor

ABSTRACT Neuroblastoma is the most common solid tumor in infants. About 25% of patients have high-risk neuroblastoma, a devastating disease with poor prognosis and few treatment options. The primary driver of high-risk neuroblastoma is the oncogene MYCN, a MYC-family transcription factor that has no druggable pockets and has long eluded drug development efforts. Recently, the protein kinase Aurora A (AurA) was shown to bind to the N-Myc protein in neuroblastoma cells and interfere with its ubiquitination by the SCF ubiquitin ligase complex, preventing N-Myc from being degraded by the proteosome. Blocking complex formation between AurA and N-Myc results in rapid N-Myc degradation and cell death in neuroblastoma cell lines. The same AurA/N-Myc complex has now been shown to drive neuroendocrine prostate cancer (NEPC), and AurA also forms a similar complex with the closely- related c-Myc protein in liver cancer. These recent discoveries point to a new paradigm for targeting Myc- family transcription factors in cancer using inhibitors that trigger structural changes in AurA that block Myc protein binding and promote Myc degradation. Our lab has recently shown that most existing AurA inhibitors, including the current clinical candidate alisertib, do not have a strong enough allosteric effect on AurA to be effective at weakening N-Myc binding. In agreement with this, alisertib has inconsistent effects on N-Myc levels in cell lines, and has not performed well in ongoing clinical trials in neuroblastoma and NEPC. The weakness in our current understanding of how AurA binds to c-Myc and N-Myc and how these interactions are affected by inhibitors represents a major impediment to this therapeutic strategy for targeting Myc-driven cancers. The goal of this project is to provide the missing molecular picture of the interactions between AurA and Myc transcription factors and how they can be modulated by inhibitor binding. We plan to use new experimental tools and approaches to define how the binding of c-Myc and N-Myc alters the conformation (shape) and dynamics (protein motion) of AurA, and to delineate the specific structural changes an inhibitor must trigger to efficiently destabilize these complexes. We will a) define the structure of the AurA/Myc complexes at atomic resolution using x-ray crystallography, magnetic resonance spectroscopies and molecular modeling, b) determine how these interactions alter AurA conformation and dynamics by tracking key structural elements of the kinase in solution, c) correlate the effects of a large panel of kinase inhibitors on AurA conformation with their ability to alter the binding affinities of N-Myc and c-Myc, and d) test the efficiency of the strongest AurA allosteric modulators in a series of N-Myc- and c-Myc-dependent cancer cell lines including neuroblastoma, NEPC and liver cancer cells. The insights will pave the way for the repurposing of existing kinase inhibitors and the development of new inhibitors as a new treatment modality for Myc-driven cancers.

抽象的 神经母细胞瘤是婴儿最常见的实体瘤。大约25％的患者患有高风险 神经母细胞瘤，一种毁灭性的疾病，预后较差，几乎没有治疗选择。主要驱动程序 高危神经母细胞瘤是癌基因MYCN，它是一种Myc家庭转录因子，没有可吸毒的因素 口袋，长期以来一直避免了药物开发工作。 最近，蛋白激酶Aurora A（Aura）显示与神经母细胞瘤中的N-Myc蛋白结合 细胞并通过SCF泛素连接酶复合物干扰其泛素化，从而防止N-Myc为 被蛋白质体降解。阻断光环和N-MYC之间的复合物形成会导致快速N-MYC 神经母细胞瘤细胞系中的降解和细胞死亡。现在显示了相同的AURA/N-MYC复合体 驱动神经内分泌前列腺癌（NEPC），Aura也形成了与紧密相似的复合物 肝癌中的相关C-MYC蛋白。这些最近的发现指出了针对MYC-的新范式 使用触发AURA结构变化的抑制剂中的癌症家庭转录因子 MYC蛋白结合并促进MYC降解。 我们的实验室最近表明，大多数现有的AURA抑制剂，包括当前的临床候选者 Alisertib，对Aura没有足够强大的变构作用，无法有效地削弱N-MYC结合。在 与此一致，Alisertib对细胞系中N-MYC水平的影响不一致，并且表现不佳 在正在进行的神经母细胞瘤和NEPC的临床试验中。我们目前对光环的理解的弱点 与C-MYC和N-MYC结合以及这些相互作用如何受抑制剂的影响代表了主要障碍 针对以MYC驱动的癌症为目标的治疗策略。 该项目的目的是提供缺少的分子图片 AURA和MYC转录因子以及如何通过抑制剂结合来调节它们。我们计划使用 新的实验工具和方法来定义C-MYC和N-MYC的结合如何改变构象 （形状）和动力学（蛋白质运动），并描绘特定的结构变化抑制剂 必须触发以有效破坏这些复合物的稳定。我们将a）定义光环/MYC的结构 使用X射线晶体学，磁共振光谱和分子，在原子分辨率下的复合物 建模，b）通过跟踪关键结构来确定这些相互作用如何改变AURA构象和动态 溶液中激酶的元素，c）将大型激酶抑制剂对光环的影响相关联 符合其改变N-MYC和C-MYC的结合亲和力的能力，d）测试效率 在一系列N-MYC和C-MYC依赖性癌细胞系中，最强的Aura变构调节剂，包括 神经母细胞瘤，NEPC和肝癌细胞。见解将为现有的重新利用铺平道路 激酶抑制剂和新抑制剂作为MYC驱动的癌症的新治疗方式。