Development of targeted gene therapy to incurable sarcoma and malignant mesothelioma

开发针对无法治愈的肉瘤和恶性间皮瘤的靶向基因治疗

• 批准号: 18591651
• 负责人: YAMAMURA Hisako
• 金额: $ 2.43万
• 依托单位: Research Institute, Osaka Medical Center for Cancer and Cardiovascular Disaeses
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591651/
• 关键词: Sarcoma; Malignant mesothelioma; Herpes virus; Gene therapy; Hypoxia

We added 57 amino acids polypeptide within the ODD sequence of HIFlalpha, including proline residue at 564, to the amino terminal of HSV-1 ICP4 ptotein. Further, we added nuclear localization signal (NLS) comprising 23 amino acids to the amino terminus of ODD-ICP4.The CMV-NLS-ODD-ICP4-IRES-LacZ-polyA fragment was inserted into the Stu I site of pKpX2 (J. Virol.62,196-205,1988) (kindly provided by Dr. Weller in Coneticut University), and was linealized by digestion with XhoI. The resulting 11.3kb targeted recombination vector, UL39-CMV-NLS-ODD-ICP4-IRES-LacZ-polyA-UL39 was co-transfected with a mutant HSV-1 DNA, d120 into Vero cells (2.5x105cells/well). We isolated a single plaque of the recombinant HSV-1.The recombinant HSV-1 preferentially replicated in the hypoxic condition (O2 1%) and showed more potent anti-tumor activity. Expression of ICP4 was also enhanced in the hypoxic condition. The recombinant HSV-1 showed prominent anti-tumor activity against human malignant mesothelioma, leiomyosarcoma and breast cancer by direct injection into the tumors.

我们在HSV-1 ICP-4蛋白的氨基末端添加了HIFl α ODD序列中57个氨基酸的多肽，包括564位的脯氨酸残基。将CMV-NLS-ODD-ICP 4-IRES-LacZ-polyA片段插入pKpX 2的Stu I位点（J.Virol.62，196 - 205，1988）（由Conevolent大学的Weller博士友情提供），并用Xho I消化使其线性化。将得到的11.3kb靶向重组载体UL 39-CMV-NLS-ODD-ICP 4-IRES-LacZ-polyA-UL 39与突变体HSV-1 DNA，d120共转染到Vero细胞中（2.5 × 105个细胞/孔）。重组HSV-1在低氧条件下（O2 1%）优先复制，并显示出更强的抗肿瘤活性。ICP 4的表达在缺氧条件下也增强。重组HSV-1经瘤内注射对人恶性间皮瘤、平滑肌肉瘤和乳腺癌均有显著的抗肿瘤活性。

项目成果

Robust CD133 expression and stem cell features of gastrointestinal stromal tumors (GIST)

胃肠道间质瘤 (GIST) 的强 CD133 表达和干细胞特征

• 发表时间: 2007
• 作者: Katsuhito Takahashi;Hisako Yamamura
• 通讯作者: Hisako Yamamura

A novel variant of calponin-targeting oncolytic HSV-1 as a potent therapeutic agent for malignant mesothelioma

钙调蛋白靶向溶瘤 HSV-1 的新变体作为恶性间皮瘤的有效治疗剂

• 发表时间: 2007
• 作者: Hisako Yamamura;Katsuhito Takahashi
• 通讯作者: Katsuhito Takahashi

Loss of smooth muscle calponin results in impaired blood vessel maturation in the tumor–host microenvironment

• DOI: 10.1111/j.1349-7006.2007.00452.x
• 发表时间: 2007-05
• 期刊: Cancer Science
• 影响因子: 5.7
• 作者: H. Yamamura;N. Hirano;H. Koyama;Y. Nishizawà;Katsuhito Takahashi

ウイルス工学を応用したがん細胞標的医薬の開発 ; 肉腫・中皮腫・がん幹細胞の標的化

利用病毒工程开发针对肉瘤、间皮瘤和癌症干细胞的靶向药物；

• 发表时间: 2007
• 作者: 高橋克仁;山村倫子
• 通讯作者: 山村倫子

ウイルス工学を応用したがん細胞標的医薬の開発;肉腫・中皮腫・がん幹細胞の標的化

利用病毒工程开发针对肉瘤、间皮瘤和癌症干细胞的靶向药物；

• 发表时间: 2007
• 作者: 高橋克仁;山村倫子
• 通讯作者: 山村倫子