Pre-clinical study of gene therapy against refractory diseases in animals.

动物难治性疾病基因治疗的临床前研究。

• 批准号: 13460140
• 负责人: OHNO Koichi
• 金额: $ 11.2万
• 依托单位: THE UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13460140/
• 关键词: dogs; cats; gene therapy; Hepatocyte growth factor; tumor; adenovirus; feline immunodeficiency virus; アデノウイルスベクター; p53; MDM2; 肝細胞増殖因子(HGF)

The object of this study is the development of gene therapy against tumors and immune-abnormalities in veterinary field. In this research period, the study of 1) basic research of gene therapy by hepatocyte growth factor (HGF), 2) construction of adenovirus vector containing canine p53 and its effects for canine tumor cell lines, 3) gene therapy for and by feline immunodeficiency virus (FIV), have been conducted.1) HGF-gene therapy for liver and renal insufficiency in dogs and cats : Feline and canine HGF cDNA were cloned and their expressions were examined in various liver diseases in dogs together with tumor transforming growth factor (TGF). Furthermore, feline and canine HGF cDNA was inserted into expression plasmid and their efficient gene delivery method was examined.2) Gene therapy for tumors in dogs and cats : Aberrations of the p53 tumor suppressor gene in various tumors in dogs was investigated together with the abnormalities of centrosome. We next constructed adenovirus vector expressing canine p53 which was cloned in our laboratory and examined its cytocidal effects on canine tumor cell lines. Furthermore, feline IL-18 cDNA was cloned and expression plasmid for canine CTLA-4 fusion protein to enhance anti-tumor immunity.3) Gene therapy for FIV infection : We developed the novel method to quantify FIV RNA in plasma and an association of plasma viral RNA load with prognosis in cats naturally infected with FIV was investigated. We also investigated the mechanism of apoptosis of FIV-infected cells and cloned feline TNF receptors. Furthermore, feline vascular endotherial growth factor (VEGF) cDNA was cloned to develop new gene therapy against FIV infection.

本研究的目的是在兽医领域开发针对肿瘤和免疫异常的基因疗法。本研究期的研究内容包括1）肝细胞生长因子（HGF）基因治疗的基础研究，2）犬p53腺病毒载体的构建及其对犬肿瘤细胞系的影响，3）猫免疫缺陷的基因治疗病毒（FIV），已进行。1）针对狗和猫的肝肾功能不全的HGF基因治疗：克隆了猫科动物和犬科动物的HGF cDNA，并在不同的环境中检查了它们的表达狗的肝脏疾病与肿瘤转化生长因子（TGF）一起。此外，将猫科动物和犬科动物的HGF cDNA插入表达质粒中，并检查了其有效的基因递送方法。2)狗和猫肿瘤的基因治疗：研究了狗的各种肿瘤中p53肿瘤抑制基因的畸变以及异常。的中心体。接下来，我们构建了表达犬p53的腺病毒载体，并在我们实验室进行了克隆，并检查了其对犬肿瘤细胞系的杀细胞作用。此外，还克隆了猫IL-18 cDNA和犬CTLA-4融合蛋白的表达质粒，以增强抗肿瘤免疫力。3) FIV感染的基因治疗：我们开发了定量血浆中FIV RNA和血浆关联的新方法。研究了自然感染 FIV 的猫的病毒 RNA 载量与预后的关系。我们还研究了 FIV 感染细胞的凋亡机制和克隆的猫 TNF 受体。此外，还克隆了猫血管内皮生长因子 (VEGF) cDNA，以开发针对 FIV 感染的新基因疗法。

项目成果

Telomere length and telomerase activity in canine mammary gland tumors.

犬乳腺肿瘤中的端粒长度和端粒酶活性。

• 发表时间: 2001
• 期刊: American Journal of Veterinary Research 62
• 作者: Yazawa;M.;et al.
• 通讯作者: et al.

Effect of an adenoviral vector that expresses the canine p53 gene on cell growth of canine osteosarcoma and mammary adenocarcinoma cell lines.

表达犬p53基因的腺病毒载体对犬骨肉瘤和乳​​腺癌细胞系细胞生长的影响。

• 发表时间: 2003
• 期刊: American Journal of Veterinary Research (印刷中)
• 作者: Yazawa;M.;et al.
• 通讯作者: et al.

Kanaya, N.: "Detection of the anti-P53 antibodies in a dog with tumor"J. Vet. Med. Sci.. 64. 973-979 (2002)

Kanaya，N.：“检测患有肿瘤的狗中的抗 P53 抗体”J。

Setoguchi, A.: "Results of hyperamplification of centrosomes in naturally developing tumors of dogs"Am. J. Vet. Res.. 62. 1134-1141 (2001)

Setoguchi, A.：“狗自然发育肿瘤中中心体过度扩增的结果”Am。

Molecular cloning of feline tumour necrosis factor receptor type I (TNFR I) and expression of TNFR I and TNFR II in lymphoid cells in cats.

猫肿瘤坏死因子受体 I 型 (TNFR I) 的分子克隆以及猫淋巴细胞中 TNFR I 和 TNFR II 的表达。

• 发表时间: 2003
• 期刊: European Journal of Immunogenetics 30
• 作者: Mizuno;T.;et al.
• 通讯作者: et al.