Basic research of malignant glioma-specific gene therapy by using very high titer retrovirus vectors

超高滴度逆转录病毒载体恶性胶质瘤特异性基因治疗的基础研究

• 批准号: 12470290
• 负责人: SHIMIZU Keiji
• 金额: $ 9.15万
• 依托单位: Kochi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470290/
• 关键词: Glioma; Retrovirus Vector; Promoter; MBP; Gene Therapy; SAGE; MAGE-E1

Gene therapy for malignant glioma was done for the first time in the United States in 1992. They implanted PA317 (mouse) packaging cells, which produced the retrovirus vectors transduced the non-specific SV40 promoter, into the tumor beds of malignant glioma patients (Ram Z, et al : Nature Med, 1997). This procedure may induce not only the immunological xeno-transplantation reactions but also the serious side-effects into the normal divided cells. Therefore, we have repeated some basic research on brain-specific gene therapy controlled suicide gene (Herpes Simplex Virus thymidine kinase ; HSVtk) with promoter expressed Myelin basic protein (MBP) gene (J Neurosci Res 36, 1993). We transduced the Polyoma ori early region into the packaging cells produced retrovirus in order to obtain the high-titer retroviral vectors (Hum Gene Ther 9, 1998). And mouse brain tumor models were completely cured by these high-titer vectors (Gene Ther 5, 1998 ; Gene Ther 8, 2001 ; Human Cell 14, 2001). The sa … More fety test these vectors by using a common marmoset (primates) was approved as a confirmation experiment of the Minister of Education, Culture, Sports, Science and Technology on December 2, 2002. Now, we perform several in vitro safety studies of these high-titer brain-specific retroviral vector-producing (packaging) cells (master cells) and cryopreserve these master cells, and then take a plan of in vivo safety examinations by using common marmosets. Finally, we are planning clinical phase I studies of gene therapy for brain tumor patients in 2004.In addition, as these malignant gliomas showed high and low MBP-expressed gene, we identified MAGE-E1 gene, as one of the candidates for glioma-related genes, by using the SAGE(serial analysis of gene expression) method (Cancer Res 61, 2001 ; Gene 277, 2001). Now, we search the promoter expressing MAGE-E1 gene, and then will try a new glioma-specific gene therapy, which vectors were transduced this new promoter. Moreover, we will research the correlation between this MAGE-E1 gene appearance and major histocompatibility complexes (MHC) of glioma patients under agreement of the patient himself, according to "Ethics indication concerning human genome and gene analysis research". Less

恶性胶质瘤的基因治疗于1992年首次在美国进行。他们将产生转导非特异性SV 40启动子的逆转录病毒载体的PA 317（小鼠）包装细胞植入恶性神经胶质瘤患者的肿瘤床中（Ram Z等：Nature Med，1997）。这种方法不仅会引起异种移植的免疫反应，而且会对正常的分裂细胞产生严重的副作用。因此，我们重复了一些关于脑特异性基因治疗控制的自杀基因（单纯疱疹病毒胸苷激酶; HSVtk）与启动子表达的髓鞘碱性蛋白（MBP）基因的基础研究（J Neurosci Res 36，1993）。为了获得高滴度的逆转录病毒载体，我们将多瘤病毒ori早期区域转导到产生逆转录病毒的包装细胞中（GeneticGene Ther 9，1998）。并且小鼠脑肿瘤模型被这些高滴度载体完全治愈（Gene Ther 5，1998 ; Gene Ther 8，2001 ; Human Cell 14，2001）。的sa ...更多信息 2002年12月2日，文部科学大臣批准了用普通的绒猴（灵长类动物）对这些载体进行的确认试验。现在，我们对这些高滴度脑特异性逆转录病毒载体生产（包装）细胞（主细胞）进行了几项体外安全性研究，并将这些主细胞冷冻保存，然后采用普通绒猴进行体内安全性检查。最后，我们计划在2004年对脑肿瘤患者进行基因治疗的临床I期研究。此外，由于这些恶性胶质瘤显示出高和低MBP表达的基因，我们通过使用SAGE（基因表达的系列分析）方法（Cancer Res 61，2001 ; Gene 277，2001）鉴定了MAGE-E1基因，作为胶质瘤相关基因的候选基因之一。目前，我们正在寻找MAGE-E1基因的启动子，并将此启动子导入载体中，尝试一种新的胶质瘤特异性基因治疗方法。此外，我们将根据“人类基因组和基因分析研究的伦理学指示”，在患者本人同意的情况下，研究该MAGE-E1基因的出现与胶质瘤患者的主要组织相容性复合体（MHC）之间的相关性。少

项目成果

Tanaka F, et al.: "Expression of polysialic acid and STX, a human polysialyltransferase, is correlated with tumor progression in non-small cell lung cancer"Cancer Res. 60. 3072-3080 (2000)

Tanaka F 等人：“聚唾液酸和 STX（一种人聚唾液酸转移酶）的表达与非小细胞肺癌的肿瘤进展相关”Cancer Res。

Tanaka F, et al.: "Expression of polysialic acid and STX, a human polysialyltransferase, is correlated with tumor progression in non-small cell lung cancer"Cancer Res.. 60. 3072-3080 (2000)

Tanaka F 等人：“聚唾液酸和 STX（一种人聚唾液酸转移酶）的表达与非小细胞肺癌的肿瘤进展相关”Cancer Res.. 60. 3072-3080 (2000)

Sasaki M, et al.: "MAGE-E1, a new member of Melanoma-associated antigen gene family and its expression in human glioma"Cancer Res. 61. 4809-4814 (2001)

Sasaki M等人：“MAGE-E1，黑色素瘤相关抗原基因家族的新成员及其在人神经胶质瘤中的表达”Cancer Res。

Ikenaka K, et al.: "Treatment of glioblastoma by direct inoculation of concentrated high titerrecombinant retrovirus carrying the herpes simplex virus thymidine kinase gene"Human cell. 14(1). 49-58 (2001)

Ikenaka K 等人：“通过直接接种携带单纯疱疹病毒胸苷激酶基因的浓缩高滴度重组逆转录病毒来治疗胶质母细胞瘤”人类细胞。

Tamura K, et al.: "Eradication of murine brain tumors by direct inoculation of concentrated high titer-recombinant retrovirus harboring herpes simplex virus thymidine kinase gene"Gene Ther. 8(3). 215-222 (2001)

Tamura K 等人：“通过直接接种含有单纯疱疹病毒胸苷激酶基因的浓缩高效价重组逆转录病毒来根除小鼠脑肿瘤”Gene Ther。