Moleculo-biological analysis for mechanism of proliferation and differentiation in glioma cells and its clinical application

我们已经针对胶质瘤和髓母细胞瘤共有的抗原制备了小鼠单抗(ONS-M21)(BR J癌症68：831-837,1993)。然后，我们与中外制药有限公司合作，成功地将他们的抗体(Mol免疫)和他们的单链抗体人源化，利用这些人源化抗体和他们的单链抗体，我们可能会开发出正确的肿瘤部位诊断，并将尝试应用于恶性胶质瘤患者的免疫治疗。目前已进入最后一步，即提取这些鼠源性单抗所识别的恶性胶质瘤相关抗原。在提取它们的抗原后，我们将确定它们的基因排列和表达机制。另一方面，我们正在研究恶性胶质瘤的基因治疗，与教授。来自冈崎国立研究所、国家生理科学研究所的IKenaka和弘。我们发表了两篇关于胶质细胞特异性启动子在胶质瘤细胞中选择性表达外源基因的论文(Jpn J Cancer Res 83：1244-1247,1992；J Neurosci Res 38：415-423,1994)。目前，我们在这些体外实验数据下，用小鼠髓鞘碱性蛋白(MBP)基因启动子来指导毒性基因的表达(J Neurosci Res 36：472-479,1993)，在小鼠胶质瘤模型中重复体内基因治疗。我们证实，如果我们能将HTK基因转导到大约25%的小鼠脑内移植的胶质瘤细胞中，所有小鼠都能通过腹腔注射GCV而康复。如果我们能按计划阐明胶质瘤相关抗原，我们将分析控制其表现的启动子，并将其应用于未来具有组织特异性的基因治疗。

We had already made mouse monoclonal antibodies (ONS-M21) for the antigens which were shared in both glioma and medulloblastoma (Br J Cancer 68 : 831-837,1993). Then we succeed in humanization of their antibodies (Mol Immunol, in press) and their single-chained antibodies in cooperation with Chugai Pharmaceutical Co.Ltd.Using these humanized antibodies and their single-chained antibodies, we might develop the correct diagnoses of tumor sites, and will try to apply to immunotherapy of malignant glioma patients. And now it is in the last steps to extract the malignant glioma-associated antigens which were recognized with these mouse monoclonal antibodies. After extraction of their antigens, we will identify their gene arrangement and mechanisms of their expression. On the other hand, we are researching about the gene therapy for malignant gliomas, with prof. Kazuhiro IKenaka who belong to Okazaki National Research Institutes, National Institute for Physiological Sciences. We published two papers about selective expression of foreign genes in glioma cells by glial-specific promoters (Jpn J Cancer Res 83 : 1244-1247,1992 ; J Neurosci Res 38 : 415-423,1994). At present, we repeats the in vivo gene therapy in mouse glioma models under these in vitro data with the mouse myelin basic protein (MBP) gene promoters to direct toxic gene expression (J Neurosci Res 36 : 472-479,1993). We confirmed that all mice recovered with intraperitoneal administration of GCV,if we could transduced HTK genes into approximately 25% of glioma cells implanted into the mouse brain.If we could elucidate the glioma-associated antigens according to our plans, we will analyze the promoters controlling their manifestation and will apply them in gene therapy with tissue-specificity in the future.