Targeted and regulable expression of transgenes in hepatic stellate cells using an adenoviral Cre-loxP system

使用腺病毒 Cre-loxP 系统在肝星状细胞中进行转基因的靶向和可调节表达

• 批准号: 16590150
• 负责人: IKEDA Kazuo
• 金额: $ 2.18万
• 依托单位: Osaka City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590150/
• 关键词: Stellate cells; liver fibrosis; adenoviral vector; collagen; Cre; loxP; 臓器線維症; TGF-beta; BMP-7; Id2; Id3

Hepatic stellate cells (HSCs) are responsible for extracellular matrix synthesis accumulation, leading to liver fibrosis. The aim of this project was to develop a regulable adenoviral transduction system in order to modulate HSC activation and ultimately to either abrogate their generation of extracellular matrix or promote apoptosis. For this purpose, we constructed two recombinant adenoviral systems ; one expressing the Cre gene under the control of specific promoters and the other containing a potent expression unit that was activated by Cre recombinase-mediated recombination to remove an upstream LoxP-flanked ‘stuffier' sequence, thereby transcribing the downstream transgene of interest (Cre-loxP system). This Cre-loxP system was analyzed as a potential method to achieve the regulated expression of specific genes in HSCs. When promoter for the collagen 1A2 gene drove Cre recombinase expression in primary quiescent rat HSC, expression of reporter gene was observed. However, in activated HSC, the collagen promoter effectively drove Cre recombinase activity, as assessed by the expression. In contrast, the expression was barely observed when the collagen promoter was expressed in hepatocytes. HSC-specific expression of Smad7 significantly reduced the expression of type I collagen in culture and decreased fibrosis in two liver fibrosis models. These results demonstrate the potential utility of transcriptionally controlled gene therapy using a Cre/loxP system to ameliorate hepatic fibrosis in vivo.

肝星细胞（HSC）负责肝脏外肿块的肿瘤，o开发可调节的腺苷酸。通过CRE重组介导的ER序列激活的下游转基因（CRE-LoxP系统）在HSC启动子中的特定基因的典型表达但是，在激活的HSC INASE活性中观察到报告基因的表达，而在肝脏中促进的胶原蛋白几乎没有观察到SMAD7的胶原蛋白。这些结果证明了使用CRE/LOXP系统到肝的转录控制基因疗法的实用性。

项目成果

Expression of Smad7 in mouse eyes accelerates healing of corneal tissue after exposure to alkali

• DOI: 10.1016/s0002-9440(10)62358-9
• 发表时间: 2005-05-01
• 期刊: AMERICAN JOURNAL OF PATHOLOGY
• 影响因子: 6
• 作者: Saika, S;Ikeda, K;Roberts, AB
• 通讯作者: Roberts, AB

Adenoviral gene transfer of BMP-7, Id2, or Id3 suppresses injury-induced epithelial-to-mesenchymal transition of lens epithelium in mice

• DOI: 10.1152/ajpcell.00306.2005
• 发表时间: 2006-01-01
• 期刊: AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY
• 影响因子: 5.5
• 作者: Saika, S;Ikeda, K;Ooshima, A
• 通讯作者: Ooshima, A

Therapeutic effect of topical administration of SN50,an inhibitor of nuclear factor-kB, in treatment of corneal alkali burns in mice

核因子kB抑制剂SN50局部给药对小鼠角膜碱烧伤的治疗作用

• 发表时间: 2005
• 期刊: Am J Pathol (In press)
• 作者: Wei M;Fujiki K;Ando E;Zhang S;Ozaki T;Ishiguro H;Kondo T;Nokihara K;Wray V;Naruse S;Saika et al.
• 通讯作者: Saika et al.

Induction of initial cardiomyocyte alpha-actin--smooth muscle alpha-actin--in cultured avian pregastrula epiblast : a role for nodal and BMP antagonist

初始心肌细胞α-肌动蛋白--平滑肌α-肌动蛋白--在培养的禽类前原肠胚外胚层中的诱导：节点和BMP拮抗剂的作用

• 发表时间: 2005
• 期刊: Dev Dyn 233・4
• 作者: 秋元 義弘;川上 速人;Matsui H
• 通讯作者: Matsui H

Inhibition of p38MAP kinase suppresses fibrotic reaction of retinal pigment epithelial cells

• DOI: 10.1038/labinvest.3700294
• 发表时间: 2005-07-01
• 期刊: LABORATORY INVESTIGATION
• 影响因子: 5
• 作者: Saika, S;Yamanaka, O;Ooshima, A
• 通讯作者: Ooshima, A