Detection and regulation of hepatic diseases targeting TGF-β activation reaction.

靶向TGF-β激活反应的肝病检测与调控。

• 批准号: 16390215
• 负责人: KOJIMA Soichi
• 金额: $ 9.02万
• 依托单位: RIKEN
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390215/
• 关键词: Fulminant hepatitis; TGF-β activation; Plasma kallikrein; Plasmin; Cutting edge recognizing antibody; Protease inhibitor; Hepatic stellate cells; Smad

Transforming growth factor (TGF)-β, a 25 kD dimeric polypeptide playing a pivotal role in the pathogenesis of liver diseases, is produced as a high molecular weight latent form, and activated by plasmin (PLN) and plasma kallikrein (PLK) during pathogenesis of liver diseases. Blockage of the activation reaction with low molecular weight protease inhibitors prevented the development of the diseases in animal models. Based upon these findings, here, we found that(1) PLN and PLK cleaved between K56-L57 and R58-L59 in LAP portion of human latent TGF-β1, respectively. We produced antibodies that specifically recognize the neo-epitopes formed by protease degradation, namely the cut ends of each cleavage site. The anti-R58 antibodies strongly stained liver sections from patients with fulminant hepatitis.(2) The anti-L59 antibodies were successively used to establish ELISA to detect LAP degradation fragments in serum of mouse impaired liver regeneration model as well as rat CCl4 and bile duct ligation hepatic fibrosis models, showing a good correlation with serum transamidase levels and hepatic hydroxyproline levels.(3) Decoy peptides containing the cleavage sites blocked the activation of TGF-β in the hepatic stellate cells and improved hepatic regeneration in the mouse model.(4) In scleroderma TGF-β2 is the main isoform and MMP, but not PLN and PLK, appeared to be involved in its activation reaction.(5) CXZ, which has been shown to suppress phosphorylation of Smad, successfully improved hepatic regeneration in the mouse model.These results suggest that a PLN/PLK-dependent activation reaction occurs around hepatic stellate cells to produce active TGF-β, which may induce fibrosis and inhibit the proliferation of hepatocytes, thereby addressing a potential use for decoy peptides in hepatitis therapy. These results also suggest a potential usage of a LAP degradate as a novel biomarker for liver fibrosis.

转化生长因子-β是一种在肝病发病机制中起关键作用的25kD二聚体多肽，以高分子量潜伏形式产生，在肝病发病过程中被纤溶酶和激肽释放酶激活。在动物模型中，用低分子量的蛋白酶抑制剂阻断激活反应可以防止疾病的发展。基于这些发现，我们发现：(1)在人潜伏的转化生长因子-β1的LAP部分，PLN和PLK分别在K56-L57和R58-L59之间裂解。我们生产的抗体专门识别由蛋白酶降解形成的新表位，即每个裂解位点的切端。用抗R58抗体对重型肝炎患者的肝组织切片进行了染色。(2)先后用抗L59抗体建立了检测肝再生受损小鼠模型和大鼠肝纤维化模型血清中LAP降解片段的酶联免疫吸附试验，发现LAP降解片段与血清转氨酶水平和肝组织羟脯氨酸水平具有良好的相关性。(3)含有切割位点的诱饵多肽阻断了肝星状细胞转化生长因子-β的激活，促进了肝再生。(4)在硬皮病模型中，转化生长因子-β-2是主要的亚型，而不是磷脂酰肌醇和磷脂酰肌醇。(5)CXZ能抑制Smad的磷酸化，成功地促进了小鼠的肝再生。这些结果表明，肝星状细胞周围发生了依赖pln/plk的激活反应，产生了活性的转化生长因子-β，从而可能诱导肝纤维化，抑制肝细胞的增殖，从而为诱骗多肽在肝炎治疗中的潜在应用提供了可能。这些结果还表明LAP降解物作为肝纤维化的一种新的生物标志物的潜在用途。

项目成果

TGF-β情報伝達経路阻害剤

TGF-β信号通路抑制剂

• 发表时间: 2003

Synergistic growth inhibition by acyclic retinoid and vitamin K2 in human hepatocellular carcinoma cells

• DOI: 10.1111/j.1349-7006.2006.00384.x
• 发表时间: 2007-03-01
• 期刊: CANCER SCIENCE
• 影响因子: 5.7
• 作者: Kanamori, Toh;Shimizu, Masahito;Moriwaki, Hisataka
• 通讯作者: Moriwaki, Hisataka

Identification, evolution, and regulation of expression of guinea pig trappin with an unusually long transglutaminase substrate domain

• DOI: 10.1074/jbc.m501678200
• 发表时间: 2005-05-27
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Furutani, Y;Kato, A;Hirose, S
• 通讯作者: Hirose, S

Te involvement of polyamines as substrates of transglutaminase in zonal different hepatocyte proliferation after partial hepatectomy.

多胺作为转谷氨酰胺酶的底物参与部分肝切除术后不同区域的肝细胞增殖。

• 发表时间: 2005
• 期刊: Biol. Pharm. Bull. 28(2)
• 作者: Koike H;Hirayama M;Yamamoto M;Ito H;Hattori N;Umehara F;Arimura K;Ikeda S;Ando Y;Nakazato M;Kaji R;Hayasaka K;Nakagawa M;Sakoda S;Matsumura K;Onodera 0;Baba M;Yasuda H;Saito T;Kira J;Nakashima K;Oka N;Sobue G.;Omori K;Ohtake.Y.et al.
• 通讯作者: Ohtake.Y.et al.

TGF-β活性化制御領域の切断面を認識する抗体

识别 TGF-β 激活控制区裂解表面的抗体

• 发表时间: 2003