MIYAO Yasuyoshi Osaka Univ Hospital, Medical Stuff, 医学部・附属病院, 医員
OHNISHI Takanori Osaka Univ Med Sch, Dept of Neurosurg, Assistant Prof, 医学部, 助手 (70233210)
YOSHIMINE Toshiki Osaka Univ Med Sch, Dept of Neurosurg, Associate Prof, 医学部, 講師 (00201046)
SHIMIZU Keiji Osaka Univ Med Sch, Dept of Neurosurg, Associate Prof, 医学部, 助教授 (50162699)
|Budget Amount *help
¥6,600,000 (Direct Cost : ¥6,600,000)
Fiscal Year 1994 : ¥3,000,000 (Direct Cost : ¥3,000,000)
Fiscal Year 1993 : ¥3,600,000 (Direct Cost : ¥3,600,000)
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 diagnose 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.
We publishied 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 t
herapy 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). Then, if we could elucidate the glioma-associated antigens accordig to our plans, we will analyze the promoters controlling their manifestation and will apply them in gene therapy with tissuespecificity in the future.
On the other hand, to understand the cellular mechanism of brain invasion by glioma cells, two molecular species of glioma-derived motility factor (GMF), GMF-I and GMF-II,have been purified to homogeneity from the serum-free conditioned medium of a highly invasive human glioma cell line, T98G,by gelatin affinity chromatography and heparin affinity-, DEAE-, hydroxyapatite-, gel permeation- and sulfopropyl high performance liquid chromatography. GMF-I and GMF-II both stimulated the migration of T98G cells in a concentration-dependent manner, and the activity of GMF-I was about 5 times as strong as that of GMF-II.C_6 glioma cells, both of which showed high invasiveness in an in vitro invasion assay with reconstituted basement membrane, Matrigel, migrated to the GMFs with great intensity, while A172 and 9L glioma cells and normal glial cells, all of which weakly infiltrated the Matrigel barrier, migrated to the GMFs with much less intensity. These results indicate that migratory response of glioma cells to the GMFs correlates well with invasiveness, suggesting an important roles of the GMFs in the process of glioma cell invasion (Biochem Biophys Res Commun 193 : 518-525,1993). Less