Study on the gene therapy of soluble-HB-EGF gene to accelerate liver regeneration.

可溶性HB-EGF基因加速肝再生的基因治疗研究

• 批准号: 11670500
• 负责人: TAMURA Shinji
• 金额: $ 2.24万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670500/
• 关键词: soluble-HB-EGF; liver regeneration; in vivo; HVJ-liposome; gene therapy; hepatocyte; growth factor; HB-EGF

We have reported that heparin-binding EGF-like growth factor (HB-EGF) is a potent mitogen of hepatocytes in vitro. The level of HB-EGF mRNA in regenerating rat liver creases rapidly after a partial hepatectomy. In vitro study showed that HB-EGF induces the expression of HGF and TGF-alpha in non-parenchymal cells and hepatocytes, respectively. To clarify the role of soluble-HB-EGF, we studied the effect of an inhibitor which inhibits the post-tanslational processing from a membrane-bind form of HB-EGF to a soluble form, using HB-EGF transgenic mouse. The inhibitor decreased the labeling index of hepatocyte, indicating that soluble-HB-EGF is important in liver regeneration. We also investigated the effect of administration of soluble-HB-EGF to mice on proliferation of hepatocyte in vivo. Recombinant soluble-HB-EGF was administered intravenously to the normal mice for 4 times at 2h-interval. The labeling index is 300-fold higher in HB-EGF administered mice than that of controls. In vivo administration of HB-EGF activated NF-kB and STAT-3 in the liver. The induction of HGF was observed in HB-EGF-administered mice liver. To develop gene therapy of soluble-HB-EGF using HVJ-liposome method, we administered the HVJ-liposome to mouse intravenously. No serious adverse effect was observed by the administration. Injection of expression plasmid containing soluble-HB-EGF gene in HVJ-liposome in the *uscle of mouse increased the plasma levels of HB-EGF.These results indicated that introduction of soluble HB-EGF gene using HVJ-liposome method could be a candidate for new approach to accelerate the liver regeneration

肝素结合EGF样生长因子（HB-EGF）是一种有效的肝细胞有丝分裂原。大鼠肝部分切除后再生肝HB-EGF mRNA水平迅速升高。体外研究表明，HB-EGF分别诱导非实质细胞和肝细胞中HGF和TGF-α的表达。为了阐明可溶性HB-EGF的作用，我们使用HB-EGF转基因小鼠研究了抑制剂对HB-EGF从膜结合形式到可溶性形式的翻译后加工的影响。抑制剂降低肝细胞的标记指数，表明可溶性HB-EGF在肝再生中是重要的。我们还研究了给予可溶性HB-EGF对小鼠体内肝细胞增殖的影响。正常小鼠静脉注射重组可溶性HB-EGF，间隔2 h，共4次。HB-EGF给药小鼠的标记指数比对照组高300倍。体内施用HB-EGF激活肝脏中的NF-kB和STAT-3。在给予HB-EGF的小鼠肝脏中观察到HGF的诱导。为了研究HVJ脂质体法介导的可溶性HB-EGF基因治疗，我们将HVJ脂质体经静脉注射给小鼠。给药期间未观察到严重不良反应。将含有可溶性HB-EGF基因的表达质粒HVJ-脂质体注射小鼠肌肉后，血浆中HB-EGF水平明显升高，提示HVJ-脂质体法导入可溶性HB-EGF基因可能是促进肝再生的一种新途径

项目成果

木曽真一 他: "肝再生および肝発癌におけるサイトカイン発現様式の変化-HB-EGFおよびTGF-βを中心として-。"肝臓. 41・1. 64-66 (2000)

Shinichi Kiso 等人：“肝脏再生和肝癌发生过程中细胞因子表达模式的变化 - 关注 HB-EGF 和 TGF-β 肝脏 64-66”。

Kiso S,Kawata S,Tamura S: "Gastroenterology & Hepatology : Millenium 2000"H. Asakura ed. Springer-Verlag Tokyo(in press). (2001)

Kiso S、Kawata S、Tamura S：“胃肠病学

Kiso S, et al.: "Expression of heparin-binding epidermal growth factor-like growth factor in the hepatocytes of fibrotic rat liver during hepatocarcinogenesis."J Gastroenterol Hepatol. 14・12. 1203-1209 (1999)

Kiso S等人：“肝癌发生期间纤维化大鼠肝脏的肝细胞中肝素结合表皮生长因子样生长因子的表达”。J Gastroenterol Hepatol 14·12（1999）。

木曽真一: "肝臓フォーラム′00"原田尚,谷川久一 編集 医事出版社(印刷中). (2001)

Shinichi Kiso：“Liver Forum 00”，由 Hisashi Harada 和 Hisaichi Tanikawa 编辑，Iji Publishing（目前正在印刷）（2001 年）。

Kiso S, et al.: "Effects of exogenous human heparin-binding epidermal growth factor-like growth factor on DNA synthesis of hepatocytes in normal mouse liver."Biochem Biophys Res Commun. 259・3. 683-687 (1999)

Kiso S 等人：“外源性人肝素结合表皮生长因子样生长因子对正常小鼠肝脏中肝细胞 DNA 合成的影响”。Biochem Biophys Res Commun. 259·3 (1999)。