Proteomic Study of Novel Platinum Compounds in Glioma

神经胶质瘤中新型铂化合物的蛋白质组学研究

• 批准号: 7069170
• 负责人: OLIVER BOGLER
• 金额: $ 14.65万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-23 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7069170
• 关键词: antineoplastics; astrocytes; biological signal transduction; biotechnology; cell death; cell line; cisplatin; drug resistance; drug screening /evaluation; gene expression; gene mutation; glioma; laboratory mouse; liquid chromatography; mass spectrometry; mitogen activated protein kinase; neoplasm /cancer chemotherapy; organometallic compounds; p53 gene /protein; pharmacokinetics; platinum; proteomics

DESCRIPTION (provided by applicant): Gliomas remain a serious public health problem with no effective treatment, making the identification of new chemotherapeutics a priority. Equally important is the timely development of molecular tools allow the stratification of patients into likely responders and non-responders, so that therapies can be targeted effectively. A new family of platinum compounds, founded by the trinuclear BBR3464, is much more potent than established platinum agents in the killing of glioma cells in vitro. The most potent compound, BBR3610 has an IC90 in a glioma cell clonogenic assay of 8 nM, as compared to 2 mM for cisplatin. We are attempting to establish molecular profiles of responding tumors, and investigating the mechanism of action of these promising drugs by various means, and in this proposal seek support for the newest approach, which is based on displaying the proteome using two-dimensional liquid chromatography. Proteins are separated in the first dimension by a chromatofocusing column and in the second dimension on a reverse phase HPLC column. Absorbance data at 214 nM is then displayed using specialized software allowing the rapid identification of expression differences. In addition to being turnkey and suited to high throughput, this approach leaves proteins entirely in the liquid phase making recovery for downstream analysis, such as mass spectrometry, easy. We propose to apply this technology to two model systems with the aim of testing the hypothesis that cell death in response to platinum compounds is associated with characteristic changes in the proteome and that the proteins involved can be used to profile patients. In the first model, we will treat glioma cell lines with isodoses of cisplatin and BBR compounds in vitro and in vivo and assess the impact on their proteomes. Sensitive and resistant isolates of the same cell lines will be compared. In the second model, we will compare the impact of p53 mutation, which can sensitize or protect cells, on the response of cells to platinum compounds at the protein level, by using transformed p53-null astrocytes engineered to express human p53 mutants. We hope to identify changes in the proteome that are characteristic of effective glioma cell killing by platinum compounds, as such information may facilitate identifying patients likely to respond to a given drug, as well as aid in understanding their mechanism of action.

描述（由申请人提供）：神经胶质瘤仍然是一个严重的公共卫生问题，没有有效的治疗方法，因此确定新的化疗药物成为当务之急。同样重要的是分子工具的及时开发可以将患者分为可能的反应者和无反应者，以便有效地进行治疗。由三核 BBR3464 建立的一个新的铂化合物家族，在体外杀死神经胶质瘤细胞方面比现有的铂制剂更有效。 BBR3610 是最有效的化合物，在神经胶质瘤细胞克隆形成试验中的 IC90 为 8 nM，而顺铂的 IC90 为 2 mM。我们正在尝试建立响应肿瘤的分子谱，并通过各种方式研究这些有前途的药物的作用机制，并在本提案中寻求对最新方法的支持，该方法基于使用二维液相色谱显示蛋白质组。蛋白质通过色谱聚焦柱在第一维上分离，并在反相 HPLC 柱上在第二维上分离。然后使用专用软件显示 214 nM 的吸光度数据，以便快速识别表达差异。除了交钥匙且适合高通量之外，这种方法还使蛋白质完全处于液相，使得下游分析（例如质谱分析）的回收变得容易。我们建议将该技术应用于两个模型系统，目的是测试以下假设：铂化合物引起的细胞死亡与蛋白质组的特征变化相关，并且所涉及的蛋白质可用于分析患者。在第一个模型中，我们将在体外和体内用等剂量的顺铂和 BBR 化合物处理神经胶质瘤细胞系，并评估对其蛋白质组的影响。将比较相同细胞系的敏感和耐药分离株。在第二个模型中，我们将通过使用经过工程设计表达人类 p53 突变体的转化 p53 缺失星形胶质细胞，比较 p53 突变（可以使细胞敏感或保护细胞）对细胞在蛋白质水平上对铂化合物的反应的影响。我们希望确定蛋白质组的变化，这些变化是铂化合物有效杀死神经胶质瘤细胞的特征，因为这些信息可能有助于识别可能对给定药物产生反应的患者，并有助于了解其作用机制。

项目成果

Analysis of glioma cell platinum response by metacomparison of two-dimensional chromatographic proteome profiles.

通过二维色谱蛋白质组图谱的元比较分析神经胶质瘤细胞铂反应。

• DOI: 10.1074/mcp.m500124-mcp200
• 发表时间: 2006
• 期刊: Molecular & cellular proteomics : MCP
• 作者: Billecke,Christine;Malik,Imran;Movsisyan,Ashley;Sulghani,Syed;Sharif,Azita;Mikkelsen,Tom;Farrell,NicholasP;Bögler,Oliver
• 通讯作者: Bögler,Oliver