Study on gene therapy for liver regeneration. -Effect of gene transduction of soluble HB-EGF and soluble TGF-β type II receptor on liver regeneration-

肝再生基因治疗研究-可溶性HB-EGF和可溶性TGF-βII型受体基因转导对肝再生的影响-

• 批准号: 13670517
• 负责人: TAMURA Shinji
• 金额: $ 1.92万
• 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: 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/grant/KAKENHI-PROJECT-13670517/
• 关键词: liver regeneration; soluble receptor; soluble HB-EGF; TGF-β; gene therapy; 可溶性TGF-βII型受容体; 可溶型HB-EGF; HVJ-リポソーム

Acceleration of liver regeneration is important for the treatment of hepatic failure. We have reported that HB-EGF is a potent mitogen of hepatocyte and plays an important role in liver regeneratian. On the other hands, TGF-β, a growth inhibitory factor stimulates apoptosis of hepatocyte and inhibits liver regeneration.In this study, we demonstrated that NF-kB is activated in hepatic non-parenchymal cells after partial hepatectomy and stimulates the expression of HB-EGF gene. We also indicated that soluble form of HB-EGF is essential for liver regeneration using HB-EGF transgenic mice.To develop the new methods for acceleration the liver regeneration, dual introduction of soluble HB-EGF gene and soluble TGF-βtype II receptor that blocks the action of TGF-β is investigated. We developed the expression plasmid containing soluble TGF-βtype II receptor cDNA attached with human immunoglobulin Fc region or soluble HB-EGF cDNA. Both genes were introduced into mice skeletal muscle using in vivo electroporation methods. The plasma concentration of soluble HB-EGF and soluble TGF-β type II receptor were measured with newly developed ELISA systems. Pretreatment with hyaluronidase increased the expression levels and obtains 6-fold higher concentrations of soluble HB-EGF or soluble TGF-βtype II receptor.Our results give a basis for development of the gene therapy to accelerate the liver regeneration.

加速肝再生对于肝衰竭的治疗很重要。我们报道HB-EGF是一种有效的肝细胞有丝分裂原，在肝脏再生中发挥重要作用。另一方面，生长抑制因子TGF-β刺激肝细胞凋亡并抑制肝再生。在本研究中，我们证明部分肝切除术后肝非实质细胞中NF-kB被激活并刺激HB-EGF基因的表达。我们还表明，可溶性形式的 HB-EGF 对于 HB-EGF 转基因小鼠的肝再生至关重要。为了开发加速肝脏再生的新方法，研究了可溶性 HB-EGF 基因和阻断 TGF-β 作用的可溶性 TGF-β II 型受体的双重引入。我们开发了含有可溶性TGF-βII型受体cDNA并附有人免疫球蛋白Fc区或可溶性HB-EGF cDNA的表达质粒。使用体内电穿孔方法将这两种基因引入小鼠骨骼肌中。使用新开发的 ELISA 系统测量可溶性 HB-EGF 和可溶性 TGF-β II 型受体的血浆浓度。透明质酸酶预处理增加了表达水平，并获得了6倍高浓度的可溶性HB-EGF或可溶性TGF-βII型受体。我们的结果为开发加速肝脏再生的基因疗法奠定了基础。

项目成果

S. Sakuda, S. Tamura, A. Yamada, J. Miyagawa, K. Yamamoto, S. Kiso, N. Ito, K. Imanaka, A. Wada, T. Naka, T. Kishimoto, S. Kawata, Y. Matsuzawa: "Activation of signal transducer and activator transcription 3 and expression of suppressor of cytokine signal

S. Sakuda、S. Tamura、A. Yamada、J. Miyakawa、K. Yamamoto、S. Kiso、N. Ito、K. Imanaka、A. Wada、T. Naka、T. Kishimoto、S. Kawata、Y.

Sakuta S, Tamura S et al.: "NF-kB activation in non-parenchymal liver cells after partial hepatectomy in rats : possible involvement in expression of heparin-binding epidermal growth factor-like growth factor"J. Hepatology. (in press). (2001)

Sakuta S、Tamura S 等人：“大鼠部分肝切除术后非实质肝细胞中 NF-kB 的激活：可能参与肝素结合表皮生长因子样生长因子的表达”J.

Sakada S, Tamura S, et al.: "NF-kappaB activation an non-parenchymal liver cells after partial hepatectomy in rats : possible involvement in expression of heparin-binding epidermal growth factor-like growth factor"J Hepatol. 36・4. 527-533 (2001)

Sakada S、Tamura S 等人：“大鼠部分肝切除术后非实质肝细胞的 NF-kappaB 激活：可能参与肝素结合表皮生长因子样生长因子的表达”J Hepatol 36・4。 527-533 (2001)

S.Sakuda, S.Tamura, et al.: "Activation of signal transducer and activator of transcription 3 and expression of suppressor of cytokine signal 1 during liver regeneration in rats"J. Hepatol.. 36. 378-384 (2002)

S.Sakuda、S.Tamura 等人：“大鼠肝再生过程中信号转导子和转录激活子 3 的激活以及细胞因子信号 1 抑制子的表达”J.

Sakuta S, Tamura S et al.: "Activation of signal transducer and activator transcription 3 and expression of suppressor of cytokine signal 1 during liver regeneration in rats"J. Hepatology. (in press). (2001)

Sakuta S、Tamura S 等人：“大鼠肝再生过程中信号转导子和激活子转录 3 的激活以及细胞因子信号 1 抑制子的表达”J.