Mechanisms of ID2 regulation in glioma

胶质瘤中ID2的调控机制

• 批准号: 10377359
• 负责人: ANNA LASORELLA
• 金额: $ 30.97万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2022-09-05
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10377359
• 关键词: Address; Affinity; Alleles; BHLH Protein; Binding; Biochemical; Biological; Biology; Brain; Cancer Biology; Cellular biology; Chemoresistance; Complex; Conceptions; Cullin 2 Protein; DNA Binding; Dissociation; Elements; Event; Family; Genes; Genetic; Genetic Transcription; Glare; Glioblastoma; Glioma; Gliomagenesis; Human; Hypoxia; Hypoxia Inducible Factor; ID2 gene; In Vitro; Individual; Inhibitor of Differentiation Proteins; Knock-in; Knock-in Mouse; Knockout Mice; Laboratories; Link; Maintenance; Malignant Glioma; Malignant Neoplasms; Mediating; Messenger RNA; Metabolic; Modeling; Molecular; Mouse Strains; Mus; Mutation; Nature; Neuroepithelial, Perineurial, and Schwann Cell Neoplasm; Normal Cell; Oncogenes; Oncoproteins; Outcome; Oxygen; Pathway interactions; Patients; Phenotype; Phosphoproteins; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Process; Procollagen-Proline Dioxygenase; Property; Protein Biochemistry; Protein phosphatase; Proteins; Regulation; Reporting; Role; Signal Transduction; Testing; Threonine; Tissues; Tumor Angiogenesis; Tumor Cell Invasion; Tumor Suppressor Proteins; Validation; Work; angiogenesis; base; cancer cell; cancer stem cell; conditional knockout; elongin B; elongin C; experimental study; gain of function; genetic signature; human disease; hypoxia inducible factor 1; in vivo; mouse genetics; mouse model; mutant; nerve stem cell; novel; predictive signature; premature; preservation; prevent; protein function; small hairpin RNA; stem cell model; stem cells; stemness; transcription factor; tumor; tumor progression; tumorigenesis; tumorigenic; ubiquitin ligase

Abstract Mechanisms that maintain cancer stem cells are crucial to tumor progression. ID proteins are essential to support stemness, tissue invasion and angiogenesis in malignant glioma and other tumor types. Among ID proteins, ID2 contributes to cancer hallmarks, promotes chemoresistance of neural tumors and is part of a gene signature that predicts poor outcome in patients with high-grade glioma. Hypoxia–Inducible Factors (HIFs), most notably HIF2, are expressed in and required for maintenance of cancer stem cells. However, the pathways that are engaged by ID2 or drive HIF2 accumulation during tumor progression have remained unclear. In this proposal, we will follow on our most recent work that has reconstructed the molecular events linking activation of ID2 and elevation of HIF2 in cancer. Our work indicates that disruption of the VHL-Elongin C-Elongin B (VCB)-Cul2 complex by un-phosphorylated ID2-Thr-27 is an important mechanism of HIF2 stabilization in stem cells and glioma stem cells (GSCs) and that ID2 activity is restrained by prolyl hydroxylase 1 (PHD1)-induced DYRK1 kinase activation and ID2-Thr-27 phosphorylation under normoxic conditions. Interestingly an ID2-T27A mutant constitutively inactivates VCB-Cul2, elevates HIF and maintains cancer stem cells. In the proposal, we will characterize biochemically and functionally PHD-DYRK1 activity, a new tumor suppressor pathway that operates by restraining the interfering effect of active ID2 on VCB-Cul2 ubiquitin ligase. In Aim 1 we will identify the ID2- pT27 phosphatase, a potential oncoprotein and elucidate the broad significance of Thr-27 phosphorylation for the ID2 interactome. The biological implication of the PHD1-DYRK1-ID2-HIF2 pathway for glioma progression will be explored in Aim 2. Using PHD1Flox mice we will mechanistically dissect the function of constitutive ID2 activation in the absence of PHD1 in vivo. We will also model VHL inactivation, HIF stabilization and gliomagenesis by ID2-T27A mutation in an inducible mouse model harboring a knock-in allele of the mutant ID2. We will use this mouse to validate the biochemical and biological effects of ID2-T27A that converge on VHL and HIF in vivo. The mouse model will also be used to determine whether ID2-T27A promotes glioma progression in cooperation with other tumor-specific mutations that are known to cooperate with ID2 in human cancer. Studies will address fundamental questions in cancer biology and greatly enhance the understanding of how hallmarks of glioma progression are effected by ID2. Finally, the significance of the ID2-VHL-HIF-axis for the human disease will be tested in patient-derived GSCs and primary GBM.

抽象的 维持癌症干细胞的机制对于肿瘤进展至关重要。 ID 蛋白对于支持至关重要 恶性神经胶质瘤和其他肿瘤类型的干性、组织侵袭和血管生成。 ID 蛋白中，ID2 有助于形成癌症特征，促进神经肿瘤的化疗耐药性，并且是基因特征的一部分 预测高级别神经胶质瘤患者的预后不良。最值得注意的是缺氧诱导因子 (HIF) HIF2α 在癌症干细胞中表达，并且是维持癌症干细胞所必需的。然而，这些途径是 在肿瘤进展过程中 ID2 参与或驱动 HIF2α 积累仍不清楚。在这个提案中， 我们将继续我们最近的工作，该工作重建了与 ID2 激活和 癌症中 HIF2α 的升高。我们的工作表明，VHL-Elongin C-Elongin B (VCB)-Cul2 的破坏 未磷酸化的 ID2-Thr-27 形成的复合物是干细胞中 HIF2α 稳定的重要机制， 胶质瘤干细胞 (GSC) 且 ID2 活性受到脯氨酰羟化酶 1 (PHD1) 诱导的 DYRK1 抑制 常氧条件下激酶激活和 ID2-Thr-27 磷酸化。有趣的是 ID2-T27A 突变体 组成型灭活 VCB-Cul2，升高 HIFα 并维持癌症干细胞。在提案中，我们将 表征 PHD-DYRK1 活性的生化和功能，这是一种新的肿瘤抑制途径 通过抑制活性ID2对VCB-Cul2泛素连接酶的干扰作用。在目标 1 中，我们将识别 ID2- pT27 磷酸酶，一种潜在的癌蛋白，阐明了 Thr-27 磷酸化的广泛意义 ID2 相互作用组。 PHD1-DYRK1-ID2-HIF2α通路对神经胶质瘤进展的生物学意义 将在目标 2 中进行探索。使用 PHD1Flox 小鼠，我们将机械地剖析组成型 ID2 的功能 在体内没有 PHD1 的情况下激活。我们还将模拟 VHL 失活、HIFα 稳定和 在含有突变 ID2 敲入等位基因的可诱导小鼠模型中，ID2-T27A 突变导致神经胶质瘤发生。 我们将使用该小鼠来验证 ID2-T27A 收敛于 VHL 和 HIF 体内。该小鼠模型还将用于确定 ID2-T27A 是否促进神经胶质瘤进展 与已知与人类癌症中的 ID2 合作的其他肿瘤特异性突变合作。研究 将解决癌症生物学的基本问题，并极大地增强对标志如何产生的理解 神经胶质瘤的进展受 ID2 的影响。最后，ID2-VHL-HIF-α轴对于人类的意义 疾病将在患者来源的 GSC 和原发性 GBM 中进行测试。