Proteomics of Pancreatic Fluid and Urine in Chronic Pancreatitis

描述（由申请人提供）：慢性胰腺炎（CP）是一种进行性衰弱疾病，只能进行中晚期诊断。此外，在怀疑CP的情况下对慢性腹痛的评估导致每年数百万美元的医疗费用和工资损失。CP的特点是慢性炎症和进行性瘢痕形成，导致胰腺不可逆损伤，导致外分泌和内分泌功能丧失，在某些情况下还会导致胰腺癌。由于我们对该病的了解不足，以及早期和轻度疾病的影像学研究中形态学变化很少，因此在“治疗窗口”内诊断CP仍然难以捉摸。了解CP发病机制对早期诊断至关重要。此外，以炎症为基础的病理生理学治疗可以改变或延缓疾病进展。我在本研究计划中讨论的与健康相关的主题是：(1)目前对CP的发病机制知之甚少；(2)CP的临床诊断只能检测到中度至晚期疾病。我假设可以通过对胰液（PF）进行全面、定量的蛋白质组学分析来鉴定cp特异性蛋白质，并且胰腺蛋白分泌组的这种扰动反映在排泄的尿液中。在特异性目标1 （SA1）中，我将确定PF中的炎症蛋白是否具有病因特异性。使用定量质谱法，我们将评估来自6种不同病因的晚期CP患者的PF，以及一个未患病的对照组，并比较炎症蛋白谱，以生成一个纳入清单，用于特异性目标2 （SA2）。该蛋白列表将在SA2中使用两个大队列进行验证(1)非胰腺疾病和(2)不同程度胰腺缺陷的CP。这些结果对于评估在SA1中发现的蛋白的诊断应用价值具有重要意义。在Specific Aim 3 （SA3）中，我将使用定向定量质谱法研究CP受试者尿液中的炎症相关蛋白。我假设在SA1中发现并在SA2中证实的蛋白质也可以在SA3的尿液中发现。使用本文概述的策略确定的CP特异性炎症相关蛋白将来自：(1)胰液-近端体液，是研究胰腺功能障碍生理途径的理想选择；(2)尿液-一种非侵入性收集的液体，非常适合临床诊断。虽然有许多种类的蛋白质可以用蛋白质组学研究PF，但我选择专注于炎症蛋白，因为它们的表达特征可能具有诊断和治疗意义。为了符合NIDDK的使命，我的目标是确定不同的CP特异性炎症蛋白的身份、相对数量、细胞起源和分子功能，其长期目标是(1)了解CP的病因炎性蛋白反应，(2)确定候选诊断标志物，(3)发现定向治疗的靶点。

DESCRIPTION (provided by applicant): Chronic pancreatitis (CP) is a progressively debilitating disease for which only moderate to advanced diagnosis is possible. In addition, the evaluation of chronic abdominal pain under the suspicion of CP results in millions of dollars in healthcare costs and lost wages annually. CP is characterized by chronic inflammation and progressive scarring leading to irreversible damage to the pancreas, resulting in loss of exocrine and endocrine function and in some cases pancreas carcinoma. The diagnosis of CP within a "therapeutic window" has remained elusive due to our poor understanding of the disease and scarce morphological changes on imaging studies in early and mild disease. The understanding of CP pathogenesis is paramount in early diagnosis. Furthermore, pathophysiology based therapy focused on inflammation could modify or retard disease progression. The health-related topics that I address in this research proposal are: (1) CP pathogenesis is currently poorly understood and (2) clinical diagnosis of CP detects only moderate to advanced disease. I hypothesize that CP-specific proteins can be identified by comprehensive, quantitative proteomic profiling of pancreatic fluid (PF) and that such perturbations of the pancreatic protein secretome is reflected in excreted urine. In Specific Aim 1 (SA1), I will determine if inflammatory proteins in PF are etiology-specific. Using quantitative mass spectrometry, we will evaluate the PF from 6 different etiologies of advanced CP, plus a non-diseased control group and compare the inflammatory protein profile to generate an inclusion list to be used in Specific Aim 2 (SA2). This list of proteins will be validated in SA2 using two large cohorts (1) non-pancreatic disease and (2) CP with varying degrees of pancreatic deficiency. These results will be important in assessing the value of the proteins discovered in SA1 for diagnostic applications. In Specific Aim 3 (SA3), I will investigate inflammation-related proteins in the urine of CP subjects using directed quantitative mass spectrometry. I hypothesize that proteins discovered in SA1 and validated in SA2 can also be identified in urine in SA3. CP- specific, inflammatory-related proteins identified using the strategy outlined herein will originate from: (1) Pancreatic fluid-a proximal body fluid which is ideal for investigation of physiological pathways of pancreatic dysfunction and (2) Urine-a noninvasively collected fluid that is well-suited for clinical diagnostics. While there are many classes of proteins that can be investigated with proteomics in PF, I chose to focus on inflammatory proteins because characterization of their expression may have both diagnostic and therapeutic implications. In conforming with the mission of the NIDDK, I aim to determine the identity, relative quantity, cellular origin, and molecular function of distinct sets of CP-specific, inflammatory proteins with the long-term goals of (1) understanding the inflammatory protein response of CP with respect to etiology, (2) identifying candidate diagnostic markers, and (3) discovering targets for directed therapy. PUBLIC HEALTH RELEVANCE: Chronic pancreatitis pathogenesis is currently poorly understood and its clinical diagnosis is limited to moderate to advanced disease. I hypothesize that etiology-specific inflammatory proteins are present in the pancreatic fluid and urine of chronic pancreatitis patients and that these proteins can be used in a mass spectrometry-based assay with diagnostic and therapeutic potential. I aim to comprehensively investigate and comparatively analyze the inflammatory proteome of non-pancreatitis and CP cohorts of different etiologies using cutting-edge and innovative mass spectrometric methodologies.