Analysis of cellular signal transduction via Neurofibromatosis tumor suppressor gene products and development of their clinical targets

通过神经纤维瘤病肿瘤抑制基因产物进行细胞信号转导的分析及其临床靶点的开发

基本信息

批准号：
19390382
负责人：
ARAKI Norie
金额：
$ 11.9万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2007
资助国家：
日本
起止时间：
2007 至 2009
项目状态：
已结题

项目摘要

Neurofibromatosis (NF) tumor suppressor gene products, neurofibromin : NF1 and merlin : NF2, are though to be important regulators of cellular growth, differentiation, and apoptosis, and implicated in the neural abnormality of NF patients. However, the precise cellular function of NF proteins has yet to be clarified. In this study, we utilized proteomic strategies, functional annotation with a proprietary gene ontology (MANGO), and standard biochemical methods to identify proteins related to neuronal differentiation in PC12 cells, which serve as a representative model system for studying NF related neuronal biological processes. Of 1,600 non-redundant proteins quantitatively identified, 72 were novel nerve growth factor-responsive PC12 proteins mostly related to the biological processes of cell morphogenesis, apoptosis/survival, and cell differentiation. Using these proteomic strategies and database, we identified a set of NF-associating cellular proteins, including axon regulator CRMP-2. These associations were crucial for the axon formation in neuron and were regulated by a series of kinase activations by CDK5, GSK-3b, and Rho kinase being controlled by neurofibromin. Our study demonstrates that the functional association of NF proteins and their associating proteins are essential for neuronal cell differentiation and that lack of expression or abnormal regulation of NF proteins can result in impaired function of neural cells, which is likely a factor in NF-related pathogenesis.

神经纤维瘤病 (NF) 肿瘤抑制基因产物，神经纤维蛋白：NF1 和 merlin：NF2，被认为是细胞生长、分化和凋亡的重要调节因子，并与 NF 患者的神经异常有关。然而，NF 蛋白的精确细胞功能尚未阐明。在这项研究中，我们利用蛋白质组学策略、专有基因本体论 (MANGO) 的功能注释和标准生化方法来鉴定 PC12 细胞中与神经元分化相关的蛋白质，作为研究 NF 相关神经元生物过程的代表性模型系统。在定量鉴定的 1,600 个非冗余蛋白中，72 个是新型神经生长因子响应性 PC12 蛋白，主要与细胞形态发生、凋亡/存活和细胞分化的生物过程相关。利用这些蛋白质组策略和数据库，我们鉴定了一组 NF 相关细胞蛋白，包括轴突调节因子 CRMP-2。这些关联对于神经元中轴突的形成至关重要，并受到 CDK5、GSK-3b 和神经纤维蛋白控制的 Rho 激酶等一系列激酶激活的调节。我们的研究表明，NF 蛋白及其相关蛋白的功能关联对于神经元细胞分化至关重要，NF 蛋白表达缺乏或异常调节可导致神经细胞功能受损，这可能是 NF 相关发病机制的一个因素。