Analysis of aberrant EGFR in malignant gliomas, and the development of immunochemotherapy using the antibody against the receptor

恶性胶质瘤中异常 EGFR 的分析以及使用针对该受体的抗体进行免疫化疗的进展

• 批准号: 04454364
• 负责人: UEDA Satoshi
• 金额: $ 4.22万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454364/
• 关键词: malignant gliomas; aberrant EGFR; survival time; antisense oligonucleotide; Lipofectin^R; gene therapy; monoclonal antibody; abenant EGFR; antiscnse oligonucleotidc; EGFR; グリオーマ(glioma)

Aberrant EGFR found in malignant gliomas, have abnormalities in the extracellular domain and intracellular domain. Aberrant EGFR was found in 14 (35%)/40 glioblastomas. Eight aberrant EGFR had an abnormality in the extracellular domain, 4 had abrromality in the intracellular domain and only 2 had abnormalities in both domains. Overepression of normal EGFR eas found in 7 glioblastomas (17.5%). In 17 anaplastic gliomas, aberrant EGFR was expressed in 3 (17.6%), but aberrant EGFR was not expressed in low grade gliomas, IN the statistical analysis of the survival time, the average survival time was 440 days in 18 patients with glioblastomas showing moderate expression of normal EGFR, 354 days in 6 patients with glioblastomas showing overexpression of normal EGFR, 318 days in 11 patients with glioblastomas demonstrating aberrant EGFR.these abnormalities may have a relationship to malignancy of gliomas whth tyrosine kinase activity. We tried an antisence EGFR oligonucleotide (D-oligonucleotide) enveloped with Lipofectin^R against three malignant glioma cell lines. The antisense EGFR oligonucleotide enveloped with Lipofectin^R inhibited the proliferation in three malignant glioma cell lines from 30 to 10% after three days incubation compared as a control incubation. The DNA synthesis activity in the supressed tumor cells dropped out about 70%. This oligonucleotide also induced accumulation of tumor cells in S and G2 + M phase. An antisense EGFR oligonucleotide enveloped with Lipofectin^R was expected to become one of the best gene therapies against malignant gliomas.We are developing the monoclonal antibody to the aberrant EGFR in order to do a diagnosis of malignant gliomas and do a immunochemotheapy against malignant gliomas.

异常EGFR存在于恶性胶质瘤中，其胞外结构域和胞内结构域均存在异常。在 40 例胶质母细胞瘤中有 14 例 (35%) 发现了异常 EGFR。 8 个异常 EGFR 在胞外结构域有异常，4 个在胞内结构域有异常，只有 2 个在两个结构域都有异常。在 7 例胶质母细胞瘤 (17.5%) 中发现了正常 EGFR 的过度抑制。 17例间变性胶质瘤中，3例（17.6%）表达异常EGFR，但低级别胶质瘤中不表达异常EGFR。在生存时间统计分析中，18例正常EGFR中度表达的胶质母细胞瘤患者平均生存时间为440天，6例正常EGFR过度表达的胶质母细胞瘤患者平均生存时间为354天，11例患者平均生存时间为318天。 表现出异常 EGFR 的胶质母细胞瘤患者。这些异常可能与具有酪氨酸激酶活性的胶质瘤恶性肿瘤有关。我们尝试用 Lipofectin^R 包裹的反义 EGFR 寡核苷酸（D-寡核苷酸）对抗三种恶性神经胶质瘤细胞系。与对照孵育相比，在三天孵育后，用Lipofectin^R包膜的反义EGFR寡核苷酸将三种恶性神经胶质瘤细胞系的增殖抑制从30%至10%。受抑制的肿瘤细胞中的 DNA 合成活性下降了约 70%。该寡核苷酸还诱导 S 期和 G2+M 期肿瘤细胞的积累。用Lipofectin^R包裹的反义EGFR寡核苷酸有望成为针对恶性胶质瘤的最佳基因疗法之一。我们正在开发针对异常EGFR的单克隆抗体，以便对恶性胶质瘤进行诊断并对恶性胶质瘤进行免疫化疗。

项目成果

須川典亮,上田聖: "Antisense DNAを用いた遺伝子治療" Clinical Neuroscience. 12(6) in press. (1994)

Noriaki Sukawa、Kiyoshi Ueda：“使用反义 DNA 进行基因治疗”《临床神经科学》12(6) 出版（1994 年）。

須川典亮: "RFLP法" 脳腫瘍の免疫と分子生物学(金芳堂). 107-121 (1992)

Noriaki Sukawa：“RFLP方法”脑肿瘤的免疫学和分子生物学（Kinhodo）107-121（1992）。

A.Janas Ekstrand, Noriaki Sugawa, C, David James, V.Peter Collins: "Amplified and rearranged epidermal growth factor receptor genes portions of the N-and/or C-terminal tails." Proc.Natl.Acad.Sci.USA. 89. 4309-4313 (1992)

A.Janas Ekstrand、Noriaki Sukawa、C、David James、V.Peter Collins：“N 端和/或 C 端尾部的表皮生长因子受体基因部分进行了扩增和重排。”

須川典亮、上田聖: "Amtisense DNAを用いた遺伝子治療" Clinical Meuroscience. 12(6)(in press). (1994)

Noriaki Sukawa、Kiyoshi Ueda：“使用 Amtisense DNA 进行基因治疗”Clinical Meuroscience 12(6)（出版中）。

Noriaki Sugawa, Satoshi Ueda, Naoya Hashimoto, Norihiro Ibayashi, Yoshio Nakagawa, Hoyoku Nihino, Kazuto Nosaka, Masanori Kurimoto: "An antisense EGFR oligonucleotide enveloped in liposome induces Growth Inhibition in human malignant gliomas" Cancer Res.

Noriaki Sukawa、Satoshi Ueda、Naoya Hashimoto、Norihiro Ibayashi、Yoshio Nakakawa、Hoyoku Nihino、Kazuto Nosaka、Masanori Kurimoto：“包裹在脂质体中的反义 EGFR 寡核苷酸可诱导人类恶性神经胶质瘤的生长抑制”Cancer Res。