Shapiro John P, Komar Hannah M, Hancioglu Baris, Yu Lianbo, Jin Ming, Ogata Yuko, Hart Phil A, Cruz-Monserrate Zobeida, Lesinski Gregory B, Conwell Darwin L
Department of Internal Medicine, Division of Gastroenterology, Hepatology, and Nutrition, Wexner Medical Center, The Ohio State University, Columbus, Ohio, USA.
Comprehensive Cancer Center, The Arthur G. James Cancer Hospital and Richard J. Solove Research Institute, Columbus, Ohio, USA.
Clin Transl Gastroenterol. 2017 Apr 13;8(4):e89. doi: 10.1038/ctg.2017.15.
Chronic pancreatitis (CP) is characterized by inflammation and fibrosis of the pancreas, leading to pain, parenchymal damage, and loss of exocrine and endocrine function. There are currently no curative therapies; diagnosis remains difficult and aspects of pathogenesis remain unclear. Thus, there is a need to identify novel biomarkers to improve diagnosis and understand pathophysiology. We hypothesize that pancreatic acinar regions contain proteomic signatures relevant to disease processes, including secreted proteins that could be detected in biofluids.
Acini from pancreata of mice injected with or without caerulein were collected using laser capture microdissection followed by mass spectrometry analysis. This protocol enabled high-throughput analysis that captured altered protein expression throughout the stages of CP.
Over 2,900 proteins were identified, whereas 331 were significantly changed ≥2-fold by mass spectrometry spectral count analysis. Consistent with pathogenesis, we observed increases in proteins related to fibrosis (e.g., collagen, P<0.001), several proteases (e.g., trypsin 1, P<0.001), and altered expression of proteins associated with diminished pancreas function (e.g., lipase, amylase, P<0.05). In comparison with proteomic data from a public data set of CP patients, a significant correlation was observed between proteomic changes in tissue from both the caerulein model and CP patients (r=0.725, P<0.001).
This study illustrates the ability to characterize proteome changes of acinar cells isolated from pancreata of caerulein-treated mice and demonstrates a relationship between signatures from murine and human CP.
慢性胰腺炎（CP）的特征是胰腺发生炎症和纤维化，导致疼痛、实质损伤以及外分泌和内分泌功能丧失。目前尚无治愈性疗法；诊断仍然困难，发病机制的某些方面仍不清楚。因此，需要识别新的生物标志物以改善诊断并了解病理生理学。我们假设胰腺腺泡区域含有与疾病过程相关的蛋白质组特征，包括可在生物流体中检测到的分泌蛋白。
使用激光捕获显微切割技术从注射或未注射雨蛙素的小鼠胰腺中收集腺泡，随后进行质谱分析。该方案能够进行高通量分析，捕获CP各个阶段蛋白质表达的变化。
鉴定出超过2900种蛋白质，而通过质谱光谱计数分析有331种蛋白质发生了≥2倍的显著变化。与发病机制一致，我们观察到与纤维化相关的蛋白质（如胶原蛋白，P<0.001）、几种蛋白酶（如胰蛋白酶1，P<0.001）增加，以及与胰腺功能减退相关的蛋白质（如脂肪酶、淀粉酶，P<0.05）表达改变。与CP患者公共数据集的蛋白质组数据相比，在雨蛙素模型组织和CP患者的蛋白质组变化之间观察到显著相关性（r=0.725，P<0.001）。
本研究阐明了对从雨蛙素处理的小鼠胰腺中分离的腺泡细胞蛋白质组变化进行表征的能力，并证明了小鼠和人类CP特征之间的关系。
