Identification of Serum Markers of Liver Fibrogenesis/ Fibrolysis by Proteomics

通过蛋白质组学鉴定肝纤维发生/纤维溶解的血清标志物

• 批准号: 7313389
• 负责人: DETLEF SCHUPPAN
• 金额: $ 21.25万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-06 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7313389
• 关键词: Accounting; Alcoholic Liver Cirrhosis; Anastomosis - action; Animals; Biliary; Biliary cirrhosis; Cessation of life; Characteristics; Cirrhosis; Collagen; Collection; Data; Deposition; Digestion; Disease regression; Enzyme-Linked Immunosorbent Assay; Evolution; Excision; Fibrosis; Future; Gene Expression; Generations; Goals; Halofuginone; Hepatic; Hepatic Fibrogenesis; Hospitals; Human; Individual; Intoxication; Label; Ligation; Liquid Chromatography; Liver; Liver Fibrosis; Liver diseases; Lobular; Mass Spectrum Analysis; Measures; Methodology; Modeling; Molecular Profiling; Monitor; Obstructive Liver Cirrhosis; Patients; Pattern; Peptides; Pharmaceutical Preparations; Prevalence; Process; Proteins; Proteome; Proteomics; Rattus; Recovery; Risk; Running; Sampling; Sampling Errors; Serum; Serum Markers; Serum Proteins; Staging; Standards of Weights and Measures; Testing; Thioacetamide; Time; Transcript; Trypsin; Validation; Week; Work; Wound Healing; base; bile duct; drug development; fibrogenesis; liver biopsy; nano; prospective; response; size; time interval

DESCRIPTION (provided by applicant): Liver fibrosis often progresses to cirrhosis. The evolution of cirrhosis is slow (decades) and monitoring of progression by frequent liver biopsies is both unethical and subject to sampling error. Fibrosis results from a dysbalance of the dynamic processes of fibrolysis (removal of matrix) in favor of fibrogenesis (deposition of matrix). There exist no noninvasive markers to measure hepatic fibrogenesis and fibrolysis. Due to the lack of such markers it has been impossible to quantify the individual risk of liver patients to progress to cirrhosis, or to develop proven antifibrotic drugs that can inhibit progression or induce fibrosis reversal. In our preliminary work we established models of progression in rats with secondary biliary cirrhosis and with panlobular cirrhosis due to thioacetamide intoxication. Cirrhosis in these animals reverses after biliodigestive anastomosis and the antifibrotic agent halofuginone, respectively. We defined specific liver gene expression profiles associated with fibrosis progression and reversal. By applying advanced serum proteomics we found first serum proteins associated with fibrogenesis and fibrolysis. We hypothesize that by using homogeneous groups of rats with progression or reversal of liver fibrosis, we can 1. relate differential serum proteomic patterns to the activity of hepatic fibrogenesis or fibrolysis as verified in the paired liver samples, and 2. identify the differentially expressed proteins. To reach these goals we pursue the following aims: 1. to thoroughly characterize the dynamics of our rat models of biliary and panlobular fibrosis progression and reversal, 2. to use quantitative proteomics with isobaric protein tags to identify serum markers of hepatic fibrogenesis and fibrolysis. To achieve these aims, we will attach 4 (8) different isobaric peptide labels (iTRAQ) to trypsin digests of 4 pools of fractionated sera from groups representing the evolution of hepatic fibrogenesis and fibrolysis after removal of abundant serum proteins. Differentially expressed proteins will be identified by 2D Nano-liquid chromatography and MALDI-TOF/TOF mass spectrometry. Based on the findings of this proposal, ELISAs for serum markers of portal vs. lobular fibrogenesis and fibrolysis will be developed in a future application. Adaptation to the human proteome and prospective validation shall allow noninvasive monitoring of fibrosis progression and regression in patients with liver diseases.

描述（由申请人提供）：肝纤维化经常发展为肝硬化。肝硬化的进化很慢（数十年），频繁的肝活检监测进展既不道德，也是遭受采样误差的影响。纤维化是由于纤维解析动态过程（去除基质）而产生的，有利于纤维化（基质的沉积）。没有用于测量肝纤维发生和纤维化的无创标记。由于缺乏这样的标记，不可能量化肝脏患者发展为肝硬化的个体风险，或开发出可以抑制进展或诱导纤维化逆转的经过验证的抗纤维化药物。 在我们的初步工作中，我们在患有次生胆道肝硬化的大鼠中建立了进展模型，并且由于硫乙酰氨酰胺中毒而引起的泛囊肝硬化。这些动物的肝硬化分别在双碘化吻合术和抗纤维化剂Halofuginone后逆转。我们定义了与纤维化进展和逆转相关的特定肝基因表达谱。通过应用晚期血清蛋白质组学，我们发现了与纤维发生和纤维分解相关的首先血清蛋白。 我们假设通过使用肝纤维化的进展或逆转大鼠的均质组，我们可以1。将差异性血清蛋白质组学模式与在成对的肝样品中验证的肝纤维发生或纤维分析的活性相关联，并确定差异表达的蛋白质。 。为了实现这些目标，我们追求以下目的：1。彻底表征我们的大鼠模型的动力学和泛细胞纤维化的进展和逆转，2。将定量蛋白质组学带有同等蛋白质标签，以识别肝纤维发生和纤维分析的血清标记。 为了实现这些目标，我们将将4（8）个不同的同醇肽标签（ITRAQ）附加到代表肝纤维发生和去除丰富的血清蛋白后的肝纤维发生和纤维化进化的4个分数血清的胰蛋白酶消化。差异表达的蛋白质将通过2D纳米液色谱和MALDI-TOF/TOF质谱法鉴定。根据该提案的发现，将在未来的应用中开发门户网站与小叶纤维发生和纤维化的血清标志物的ELISA。适应人类蛋白质组和前瞻性验证，应允许对肝病患者的纤维化进展和消退进行非侵袭性监测。