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: 10593935
• 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/10593935
• 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; Data; Development; Disease; Electrons; Elements; Eukaryotic Cell; Exhibits; Family; Fluorescence Resonance Energy Transfer; Goals; Human; In Vitro; Infant; 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; 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; programs; 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中最常见的实体瘤。约25%的患者属于高危人群 神经母细胞瘤是一种毁灭性的疾病，预后不良，治疗选择很少。主要驱动力 高危神经母细胞瘤是致癌基因 MYCN，这是一种 MYC 家族转录因子，没有可药物治疗的 口袋里，并长期逃避药物开发努力。 最近，蛋白激酶 Aurora A (AurA) 被证明可以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，防止 N-Myc 被 被蛋白酶体降解。阻断 AurA 和 N-Myc 之间的复合物形成可导致 N-Myc 快速生成 神经母细胞瘤细胞系中的降解和细胞死亡。现在已经显示了相同的 AurA/N-Myc 复合物 驱动神经内分泌前列腺癌 (NEPC)，AurA 也与密切相关的细胞形成类似的复合物 肝癌中相关的c-Myc蛋白。这些最近的发现指出了针对 Myc 的新范例 癌症中的家族转录因子使用抑制剂触发 AurA 的结构变化，从而阻断 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药物 alisertib 对 AurA 没有足够强的变构作用，无法有效削弱 N-Myc 结合。在 与此一致的是，alisertib 对细胞系中 N-Myc 水平的影响不一致，并且表现不佳 正在进行的神经母细胞瘤和 NEPC 临床试验。我们目前对 AurA 的理解存在缺陷 与 c-Myc 和 N-Myc 结合，抑制剂如何影响这些相互作用是一个主要障碍 这种针对 Myc 驱动的癌症的治疗策略。 该项目的目标是提供缺失的分子之间相互作用的图像 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 AurA 的（形状）和动力学（蛋白质运动），并描述抑制剂的特定结构变化 必须触发才能有效地破坏这些复合物的稳定性。我们将 a) 定义 AurA/Myc 的结构 使用 X 射线晶体学、磁共振波谱和分子分析技术在原子分辨率下合成配合物 建模，b) 通过跟踪关键结构确定这些相互作用如何改变 AurA 构象和动力学 溶液中激酶的元素，c) 将一大组激酶抑制剂对 AurA 的影响关联起来 构象及其改变 N-Myc 和 c-Myc 结合亲和力的能力，以及 d) 测试 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。这些见解将为重新利用现有的 激酶抑制剂和新抑制剂的开发作为 Myc 驱动的癌症的新治疗方式。