MYCN-induced calcium and magnesium signaling regulates Neuroblastoma progression

MYCN 诱导的钙镁信号传导调节神经母细胞瘤进展

• 批准号: 8544421
• 负责人: Dana-Lynn Takeko Koomoa
• 金额: $ 13.49万
• 依托单位: UNIVERSITY OF HAWAII AT HILO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-12 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8544421
• 关键词: Actins; Address; African American; Alaskan Native American; Biological Markers; Calcium; Calcium Channel; Calcium Signaling; Calcium ion; Cancer Biology; Cells; Cephalic; Cytoskeletal Modeling; Data; Development; Diagnosis; Dietary intake; Disease-Free Survival; Down-Regulation; Event; Focal Adhesion Kinase 1; Focal Adhesions; Gene Amplification; Goals; Hawaiian population; High Prevalence; Homeostasis; Integrins; Ion Channel; Knowledge; Lead; Literature; Lymphoma; MYCN gene; Magnesium; Malignant - descriptor; Malignant Childhood Neoplasm; Malignant Neoplasms; Malignant neoplasm of prostate; Measures; Mediating; Metabolism; Methodology; Methods; Modeling; Molecular; Monitor; Neuroblastoma; Patients; Pediatric Neoplasm; Peer Review; Pharmaceutical Preparations; Phosphorylation; Process; Prognostic Marker; Proteins; Proto-Oncogenes; Publications; RNA Interference; Recycling; Regulation; Research; Research Personnel; Research Support; Resistance; Role; Signal Transduction; Small Interfering RNA; Staging; Survival Rate; Time; Transcriptional Regulation; Treatment Failure; alternative treatment; cell growth regulation; cell motility; chemotherapeutic agent; effective therapy; ethnic minority population; experience; extracellular; health disparity; high risk; improved; innovation; magnesium ion; malignant breast neoplasm; migration; neuroblastoma cell; novel; outcome forecast; therapy development; treatment strategy; tumor progression

DESCRIPTION (provided by applicant): Neuroblastoma (NB) is an extra-cranial pediatric cancer. Amplification of the proto-oncogene MYCN is associated with advanced stage, high risk NB and poor prognosis. Novel drugs and alternative treatments are being investigated for patients with advanced NB with MYCN amplification. However, finding an effective treatment strategy for advanced stage NB remains elusive. In order to address this limitation, the proposed project examines the effects of calcium and magnesium signaling on cell migration and invasion in NB cells with different MYCN status. The preliminary data indicate that MYCN initiates a change in calcium signaling, accompanied by a change in the transcriptional regulation of calcium channels, namely Orai1, Orai3 and TRPM7. In addition, MYCN induces calcium-dependent NB cell migration. These observations support the hypothesis that MYCN alters calcium and magnesium homeostasis during cell migration and invasion through a mechanism that involves calcium-mediated regulation of integrin signaling and focal adhesion turnover, and calcium- and magnesium-permeable ion channels. The research plan has the following specific aims: (1) Examine the effect of MYCN on [Ca2+] and [Mg2+] homeostasis in NB cells. NB cells with different MYCN status will be cultured in external media with different extracellular [Ca2+]/[Mg2+] ratios. The intracellular calcium and magnesium will be measured, and migration and invasion rates will be determined in these cells. (2) Elucidate the mechanism by which MYCN regulation of calcium and magnesium promotes NB migration and invasion. The effect of calcium and magnesium on the expression and phosphorylation of focal adhesion kinase and integrin, focal adhesion turnover rate (assembly, disassembly and recycling of focal adhesion proteins), and actin cytoskeletal re-organization will be examined in NB cells that are actively migrating. (3) Identify the molecular components regulating NB migration and invasion. The calcium- permeable ORAI channels and the calcium- and magnesium-permeable TRPM7 channel protein will be modulated pharmacologically or using molecular methods (RNAi or transient over-expression), and the effect of these changes on cell migration and invasion, focal adhesion turnover (assembly, disassembly, and recycling of focal adhesion proteins), and intracellular [Ca2+] will be determined in NB cells. This study may identify new biomarkers for advanced stage NB, and reveal novel targets for the development of more effective chemotherapeutic drugs. In addition, this study may lead to a new model of the regulation of NB progression, which includes monitoring the status of the prognostic marker MYCN as well as the dietary intake, metabolism and cellular regulation of calcium and magnesium. This new model may apply to other cancers with MYC amplification (e.g. breast and prostate cancers, and lymphoma, etc).

描述（由申请人提供）：神经母细胞瘤（NB）是一种颅外儿科癌症。原癌基因 MYCN 的扩增与晚期、高风险 NB 和不良预后相关。正在研究针对 MYCN 扩增的晚期 NB 患者的新药和替代疗法。然而，寻找晚期 NB 的有效治疗策略仍然难以实现。为了解决这一局限性，该项目研究了钙和镁信号传导对不同 MYCN 状态的 NB 细胞迁移和侵袭的影响。初步数据表明，MYCN 启动钙信号传导的变化，伴随着钙通道转录调控的变化，即 Orai1、Orai3 和 TRPM7。此外，MYCN 还可诱导钙依赖性 NB 细胞迁移。这些观察结果支持这样的假设：MYCN 通过一种机制改变细胞迁移和侵袭过程中的钙和镁稳态，该机制涉及钙介导的整联蛋白信号传导和粘着斑周转以及钙和镁渗透性离子通道的调节。该研究计划有以下具体目标：（1）考察MYCN对NB细胞[Ca2+]和[Mg2+]稳态的影响。具有不同MYCN状态的NB细胞将在具有不同细胞外[Ca2+]/[Mg2+]比率的外部培养基中培养。将测量细胞内的钙和镁，并确定这些细胞中的迁移和侵袭率。 (2)阐明MYCN调控钙、镁促进NB迁移和侵袭的机制。将在活跃迁移的 NB 细胞中检查钙和镁对粘着斑激酶和整合素的表达和磷酸化、粘着斑周转率（粘着斑蛋白的组装、分解和再循环）以及肌动蛋白细胞骨架重组的影响。 (3)鉴定调节NB迁移和侵袭的分子成分。钙渗透性 ORAI 通道和钙镁渗透性 TRPM7 通道蛋白将通过药理学或分子方法（RNAi 或瞬时过表达）进行调节，这些变化对细胞迁移和侵袭、粘着斑周转（粘着斑蛋白的组装、分解和再循环）以及细胞内 [Ca2+] 的影响将在 NB细胞。这项研究可能会确定晚期 NB 的新生物标志物，并揭示开发更有效化疗药物的新靶点。此外，这项研究可能会带来一种调节NB进展的新模型，其中包括监测预后标志物MYCN的状态以及钙和镁的饮食摄入、代谢和细胞调节。这种新模型可能适用于其他具有 MYC 扩增的癌症（例如乳腺癌、前列腺癌、淋巴瘤等）。