Pancreatic Fluid Proteomics in Alcohol Induced Chronic Pancreatitis

描述(申请人提供)：慢性胰腺炎是一种痛苦的疾病，其特征是纤维化和正常胰腺结构的破坏。酒精滥用是慢性胰腺炎最常见的病因，占慢性胰腺炎病例的70%-80%以上。需要对酒精所致胰腺损伤的发病机制有更深入的了解：1)评估旨在改变或延缓疾病进展的治疗的益处，2)识别有长期并发症风险的患者，如吸收不良和糖尿病，以及3)协助安全和准确地评估腹痛综合征，这些症状经常被误认为是慢性胰腺炎。本研究的目的是研究酒精对慢性酒精性胰腺炎不同阶段胰液蛋白质组分泌谱的影响。对任何复杂体液的全球蛋白质组的分析都是具有挑战性的，但新兴的蛋白质组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至关重要的作用。胰液是鉴定新蛋白的极佳标本，因为与血清相比，胰液的复杂性较低，并且可能是慢性胰腺炎中可能上调或下调的低丰度蛋白的丰富来源。我们计划检验以下总体假设：酒精导致胰腺液体蛋白图谱的变化，反映了逐渐形成的疤痕、炎症和纤维化，从而导致最终的器官损害，如吸收不良、糖尿病和癌症。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窥镜采集程序，建立了一种稳健的制备方法和一种改进的样本处理技术，以在胰液中产生可重复的蛋白质图谱。具体目的：(1)建立酒精性慢性胰腺炎蛋白质组学研究的队列，收集胰液。(2)测定正常对照组、酒精性急性胰腺炎和慢性酒精性胰腺炎患者胰液蛋白质谱。从每个队列收集的胰液的完整蛋白质组将使用最先进的蛋白质组技术来确定。从定性和定量的差异、分子功能和细胞来源等方面进行比较，以探讨酒精对胰液蛋白质组分泌谱的影响。意义：确定胰液中与慢性胰腺炎疾病严重程度/功能障碍相关的独特蛋白质模式可能有助于阐明酒精导致胰腺损伤的机制。 与公共卫生相关：我们假设酒精会导致胰液蛋白图谱的改变，这些改变反映了导致最终器官损害的进行性瘢痕形成、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窥镜采集程序，建立了一种稳健的制备方法和一种改进的样本处理技术，以在胰液中产生可重复的蛋白质图谱。在胰液中识别与慢性酒精所致胰腺疾病严重程度相关的独特蛋白质模式可能有助于阐明酒精所致胰腺损伤的机制。

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.