Pancreatic Fluid Proteomics in Alcohol Induced Chronic Pancreatitis

描述（由申请人提供）：慢性胰腺炎是一种疼痛性疾病，其特征是正常胰腺结构的纤维化和破坏。酒精滥用是慢性胰腺炎最常见的病因，导致超过70- 80%的病例。需要更好地了解酒精诱导的胰腺损伤的发病机制，以：1）评估旨在改善或延缓疾病进展的治疗的益处，2）识别有长期并发症（如吸收不良和糖尿病）风险的患者，3）协助安全准确地评估经常被误解为慢性胰腺炎的腹痛综合征。本研究的目的是探讨酒精对慢性酒精性胰腺炎不同阶段胰液蛋白质组分泌的影响。任何复杂体液的全局蛋白质组分析都具有挑战性，但新兴的蛋白质组学技术在机制研究和寻找有用的疾病生物标志物方面发挥着重要作用。由于与血清相比，胰液的复杂性较低，并且可能是在慢性胰腺炎中可能上调或下调的低丰度蛋白质的丰富来源，因此胰液是用于鉴定新蛋白质的极好标本。我们计划测试以下总体假设：酒精诱导胰腺液蛋白质谱的变化，反映了导致终末器官损伤（如吸收不良、糖尿病和癌症）的进行性瘢痕形成、炎症和纤维化。为了验证我们的假设并实现我们的目标，我们开发了一种几乎无风险的内镜采集程序，建立了一种稳健的制备方法和一种精细的样品处理技术，以在胰液中生成可重现的蛋白质谱。具体目的：（1）建立队列并收集胰腺液用于酒精诱导的慢性胰腺炎的蛋白质组学分析。(2)确定健康对照、酒精诱导的急性胰腺炎和酒精诱导的慢性胰腺炎患者的胰液蛋白质谱。将使用最先进的蛋白质组学技术测定每个队列采集的胰液的完整蛋白质组。将比较所鉴定蛋白质的定性和定量差异、分子功能和细胞来源，以研究酒精对胰液蛋白质组分泌谱的影响。重要性：确定与慢性胰腺炎疾病严重程度/胰腺液功能障碍相关的独特蛋白质模式可能会阐明酒精诱导胰腺损伤的机制。 公共卫生相关性：我们假设酒精诱导胰腺液蛋白质谱的变化，反映了导致终末器官损伤的进行性瘢痕形成、炎症和纤维化。为了验证我们的假设并实现我们的目标，我们开发了一种几乎无风险的内镜采集程序，建立了一种稳健的制备方法和一种精细的样品处理技术，以在胰液中生成可重现的蛋白质谱。在胰液中识别与慢性酒精诱导的胰腺疾病严重程度相关的独特蛋白质模式可能阐明酒精诱导的胰腺损伤的机制。

DESCRIPTION (provided by applicant): Chronic pancreatitis is a painful disorder characterized by fibrosis and destruction of the normal pancreatic architecture. Alcohol abuse is the most common etiologic factor of chronic pancreatitis, resulting in over 70- 80% of cases. An improved understanding of the pathogenesis of alcohol induced pancreatic injury are needed to: 1) assess the benefits of therapy directed at modifying or retarding disease progression, 2) identify patients at risk for long term complications such as malabsorption and diabetes and to 3) assist in the safe and accurate evaluation of abdominal pain syndromes often misinterpreted as chronic pancreatitis. The objective of this proposal is to study the effects of alcohol on the proteomic secretory profile of pancreatic fluid in various stages of chronic alcohol induced pancreatitis. The analysis of the global proteome of any complex body fluid is challenging, but emerging proteomic technologies are playing an essential role in mechanistic studies and in the search for useful biomarkers of disease. Pancreatic fluid is an excellent specimen for the identification of novel proteins since it has a low complexity compared to serum and may be a rich source of low-abundant proteins which are potentially up- or down regulated in chronic pancreatitis. We plan to test the following overall HYPOTHESIS: Alcohol induces changes in the pancreas fluid protein profile that reflect the progressive scarring, inflammation and fibrosis that leads to end organ damage such as malabsorption, diabetes and cancer. To test our hypothesis and achieve our objective we have developed a virtually risk-free endoscopic collection procedure, established a robust preparation method and a refined sample handling technique to generate reproducible protein profiles in pancreatic fluid. SPECIFIC AIMS: (1) To Develop Cohorts and Collect Pancreatic Fluid for Proteomic Profiling of Alcohol- induced Chronic Pancreatitis. (2) To Determine the Protein Profile of Pancreatic Fluid in Healthy Controls, Alcohol-induced Acute Pancreatitis, and Patients with Alcohol-induced Chronic Pancreatitis. The complete proteome of collected pancreatic fluid from each cohort will be determined using state-of-the-art proteomic technology. The qualitative and quantitative differences, molecular function and cellular origin of identified proteins will be compared to investigate the effect of alcohol on the proteomic secretory profile of pancreatic fluid. SIGNIFICANCE: Identifying the unique protein patterns associated with chronic pancreatitis disease severity / dysfunction within the pancreatic fluid may illuminate the mechanisms of alcohol induced pancreas injury. PUBLIC HEALTH RELEVANCE: We hypothesize that alcohol induces changes in the pancreas fluid protein profile that reflect the progressive scarring, inflammation and fibrosis that leads to end organ damage. To test our hypothesis and achieve our objective we have developed a virtually risk-free endoscopic collection procedure, established a robust preparation method and a refined sample handling technique to generate reproducible protein profiles in pancreatic fluid. Identifying the unique protein patterns associated with chronic alcohol induced pancreatic disease severity within the pancreatic fluid may illuminate the mechanisms of alcohol induced pancreas injury.