Treatment Strategy for Organ Fibrosis Targeting TGF-β and Smad Signaling

针对 TGF-β 和 Smad 信号传导的器官纤维化治疗策略

• 批准号: 16590636
• 负责人: INAGAKI Yutaka
• 金额: $ 2.24万
• 依托单位: Tokai 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-16590636/
• 关键词: Organ fibrosis; Hepatic fibrogenesis; Gene transfer; TGF-β; Smad; Collagen; Transcriptional regulation

Background & Aims : TGF-β and its intracellular mediators, Smad proteins, play important roles in stimulating collagen gene transcription, and thus could be the targets for treating organ fibrosis. However, intervention of the TGF-β/Smad signal affects physiological signal transduction as well, and may cause serious adverse effects upon clinical application. We have attempted to suppress liver fibrosis by expressing a TGF-β/Smad antagonist selectively in collagen-producing cells only in the fibrotic liver. Methods : Recombinant adenoviruses expressing either GFP or a TGF-β/Smad signal repressor, YB-1, were injected to the mice untreated or treated with carbon tetrachloride (CCl_4). GFP fluorescence was analyzed under a confocal laser-scanning microscopy. Anti-fibrotic effects of YB-1 overexpression were examined by luciferase assays and histological examination using transgenic reporter mice. Results : When using the CAG expression unit as a control, GFP was strongly expressed in a large number of hepatocytes in both normal and CCl_4-treated liver. In contrast, GFP expression driven by a tissue-specific enhancer of the mouse α2(I) collagen gene (COL1A2) was detected in activated hepatic stellate cells in CCl_4-induced fibrotic liver, but not in untreated normal liver. There was no GFP fluorescence observed in any other organs when using the COL1A2 enhancer. Adenovirus-mediated YB-1 expression under the control of the COL1A2 enhancer significantly decreased COL1A2 promoter activity following CCl_4 injection and subsequently suppressed the progression of liver fibrosis. Conclusions : These results validate a new concept of the therapy for hepatic fibrosis to achieve cell type-specific gene expression only in the fibrotic liver with little damage to other organs.

背景和目标：TGF-β及其胞内介质Smad蛋白在刺激胶原基因转录中发挥重要作用，可能成为治疗器官纤维化的靶点。然而，TGF-β/Smad信号的干预也会影响生理信号转导，并可能对临床应用造成严重的不良影响。我们已经尝试通过仅在纤维化肝脏中的胶原产生细胞中选择性地表达TGF-β/Smad拮抗剂来抑制肝纤维化。研究方法：将表达GFP或TGF-β/Smad信号抑制子YB-1的重组腺病毒注射到未经处理或经四氯化碳（CCl_4）处理的小鼠中。在共聚焦激光扫描显微镜下分析GFP荧光。使用转基因报告小鼠通过荧光素酶测定和组织学检查来检查YB-1过表达的抗纤维化作用。结果如下：当使用CAG表达单位作为对照时，GFP在正常和CCl_4处理的肝中的大量肝细胞中强烈表达。相反，在CCl_4诱导的肝纤维化中，在活化的肝星状细胞中检测到由小鼠α2（I）胶原基因的组织特异性增强子（COL 1A 2）驱动的GFP表达，而在未处理的正常肝中未检测到。当使用COL 1A 2增强子时，在任何其他器官中均未观察到GFP荧光。在COL 1A 2增强子的调控下，腺病毒介导的YB-1表达显著降低了CCl_4注射后COL 1A 2启动子的活性，随后抑制了肝纤维化的进展。结论：这些结果证实了肝纤维化治疗的新概念，即仅在纤维化肝脏中实现细胞类型特异性基因表达，而对其他器官的损害很小。

项目成果

Multiple Proteins are involved in the protein-DNA complex in the Proximal promoter of the human α1 (III) collagen gene (COL3A1)

人类 α1 (III) 胶原蛋白基因 (COL3A1) 近端启动子中的蛋白质-DNA 复合物涉及多种蛋白质

• 发表时间: 2005
• 期刊: Biochimica et Biophysica Acta. 1729
• 作者: Matsuoka K;et al.;Goto M et al.;Fugimoto N et al.;Laub F.et al.;Yamaguchi K et al.;Yoshino T. et al.
• 通讯作者: Yoshino T. et al.

Early response of α2(I) collagen to acetaldehyde in human hepatic stellate cells is TGF-β independent

• DOI: 10.1002/hep.20798
• 发表时间: 2005-08-01
• 期刊: HEPATOLOGY
• 影响因子: 13.5
• 作者: Svegliati-Baroni, G;Inagaki, Y;Rojkind, M
• 通讯作者: Rojkind, M

肝線維化とその制御.Annual Review消化器

肝纤维化及其控制。胃肠年度评论

• 发表时间: 2005
• 作者: Yukimoto Ishii;Tadatoshi Takayama;Satoshi Asai.;稲垣 豊
• 通讯作者: 稲垣 豊

日本消化器病学会総会 2005-モノグラフ-

日本胃肠病学会会员大会2005年-专着-

• 发表时间: 2005
• 作者: Matsui;T.et al.;稲垣 豊;Mine T;稲垣 豊
• 通讯作者: 稲垣 豊

Annual Review 2005消化器

2005 年胃肠病学年度回顾

• 发表时间: 2005
• 作者: Ohkusa T;Maekawa T;Arakawa T;Nakajima M;Fujimoto K;Hoshino E;Mitachi Y;Hamada S;Mine T.Kawahara Y;Nagai T;Aoyama N;Yoshida N;Tadokoro K;Chida N;Konda Y;Seno H;Shimatani T;Ino-ue M;Sato N;稲垣 豊
• 通讯作者: 稲垣 豊