Connective Tissue Growth Factor in Hepatic Fibrosis

肝纤维化中的结缔组织生长因子

• 批准号: 6509398
• 负责人: DAVID R BRIGSTOCK
• 金额: $ 34.31万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6509398
• 关键词: acetaldehyde; adeno associated virus group; alcoholic liver cirrhosis; cell adhesion; connective tissue; extracellular matrix; fibrogenesis; fibrosis; gene delivery system; laboratory mouse; laboratory rat; liver cirrhosis; northern blottings; polymerase chain reaction; protein biosynthesis; protein structure; tissue /cell culture; transfection /expression vector; transforming growth factors; tumor necrosis factor alpha; western blottings

The broad long-term objectives are to establish the role of connective tissue growth factor (CTGF) in causing fibrotic disease. CTGF is a highly pro-fibrogenic molecule which is over- expressed in all fibrotic lesions examined to date. It is transcriptionally activated by transforming growth factor-beta (TGF-beta) and mediates many of the matrix-inducing properties that have previously been attributed to TGF-beta. The studies described in this proposal focus on the role of CTGF in liver fibrosis, including that related to alcohol abuse. Preliminary data show that CTGF is over-expressed in fibrotic livers and is produced by hepatic stellate cells (HSCs), the principal fibrogenic cell type, both in response to TGF-beta and as a function of activation. HSCs show enhanced adhesion and levels of alpha smooth muscle actin in response to CTGF. In addition, ethanol and its fibrogenic metabolite, acetaldehyde, stimulate CTGF transcription in fibroblasts. Our hypothesis is that local up-regulation of CTGF in the liver drives the fibrogenic response, including that initiated by alcohol. Our Specific Aims are (1) To determine mechanisms of CTGF regulation in HSCs, including the role played by TGF-beta and acetaldehyde (which stimulate HSCs and CTGF production) as well as retinoic acid and TNF-alpha (which inhibit HSC function and CTGF production); (2) To determine the effects of CTGF on HSC function by examining HSC DNA synthesis, division, matrix metabolism, vitamin A content, and adhesion in HSCs treated with or over-expressing various mass forms (1OkDa, 16-20kDa, 38kDa) of CTGF which occur naturally in vivo and which are a product of HSCs maintained in vitro, and to determine the role of CTGF-stimulated kinases in these processes; and (3) To produce recombinant adeno-associated viruses for the delivery of the CTGF gene into the liver in vivo to directly establish the ability of 10 kDa and 38kDa CTGF to stimulate liver fibrosis. These studies will define the fibrogenic properties of CTGF in terms of its regulation, biological properties, signaling mechanisms and protein structure. In addition, these studies will help establish whether CTGF is a therapeutic target for treating fibrosis, which is a contributing factor in 45 percent of deaths in the USA.

广泛的长期目标是确定结缔组织生长因子（CTGF）在引起纤维化疾病中的作用。 CTGF是一种高度促纤维化的分子，其在迄今为止检查的所有纤维化病变中过度表达。 它通过转化生长因子-β (TGF-β) 进行转录激活，并介导许多先前归因于 TGF-β 的基质诱导特性。 该提案中描述的研究重点关注 CTGF 在肝纤维化中的作用，包括与酗酒相关的肝纤维化。 初步数据显示，CTGF 在纤维化肝脏中过度表达，由主要的纤维化细胞类型肝星状细胞 (HSC) 产生，既响应 TGF-β，又作为激活功能。 HSC 对 CTGF 的反应显示出粘附力和 α 平滑肌肌动蛋白水平增强。 此外，乙醇及其纤维生成代谢物乙醛可刺激成纤维细胞中的 CTGF 转录。 我们的假设是，肝脏中 CTGF 的局部上调会驱动纤维化反应，包括酒精引发的纤维化反应。 我们的具体目标是 (1) 确定 CTGF 在 HSC 中的调节机制，包括 TGF-β 和乙醛（刺激 HSC 和 CTGF 产生）以及视黄酸和 TNF-α（抑制 HSC 功能和 CTGF 产生）所起的作用； (2)通过检查用体内天然存在的和体外维持的HSC产物的各种CTGF质量形式(10kDa、16-20kDa、38kDa)处理或过表达的HSC中的HSC DNA合成、分裂、基质代谢、维生素A含量和粘附来确定CTGF对HSC功能的影响，并确定CTGF对HSC功能的影响。 这些过程中 CTGF 刺激的激酶； (3)产生重组腺相关病毒，用于将CTGF基因递送到体内肝脏中，以直接建立10kDa和38kDa CTGF刺激肝纤维化的能力。 这些研究将从调节、生物学特性、信号传导机制和蛋白质结构方面定义 CTGF 的纤维化特性。 此外，这些研究将有助于确定 CTGF 是否是治疗纤维化的治疗靶点，纤维化是美国 45% 死亡的一个促成因素。