Abstract
OBJECTIVES: The secretin-stimulated endoscopic pancreatic function test (ePFT) allows for the safe collection of gastroduodenal and pancreatic fluid from the duodenum. We test the hypothesis that these endoscopically collected fluids have different proteomes. As such, we aim to show that the ePFT method can be used to collect fluid enriched in pancreatic proteins to test for pancreatic function. METHODS: Gastroduodenal and pancreatic fluid were collected sequentially from chronic pancreatitis patients undergoing an ePFT. Proteins from each fluid type were extracted using previously published optimized methods and subjected to GeLC-MS/MS analysis for protein identification and bioinformatics analysis. RESULTS: Mass spectrometry analysis identified proteins that were exclusive in either gastroduodenal (46) or pancreatic fluid (234). Subsequent quantitative analysis revealed proteins that were differentially abundant with statistical significance. As expected, proteolytic enzymes and protease inhibitors were among the differentially detected proteins. The proteases pepsinogens and gastrin were enriched in gastroduodenal fluid, while common pancreatic enzymes (e.g., aminopeptidase N, chymotrypsin C, elastase-3A, trypsin, and carboxypeptidase A1, and elastase 2B) were found in greater abundance in pancreatic fluid. Similarly for protease inhibitors, members of the cystatin family were exclusive to gastroduodenal fluid, while serpins A11, B4, and D1 were exclusive to pancreatic fluid. CONCLUSIONS: We have shown that ePFT collection coupled with mass spectrometry can be used to identify differentially detected proteins in gastroduodenal and pancreatic fluids. The data obtained using GeLC-MS/MS techniques provide further evidence supporting the feasibility of using ePFT-collected fluid to study specific diseases of the upper gastrointestinal tract, such as chronic pancreatitis.
| Original language | English |
|---|---|
| Journal | Clinical and Translational Gastroenterology |
| Volume | 3 |
| DOIs | |
| State | Published - 2012 |
Bibliographical note
Funding Information:Acknowledgements. Funds were provided by the following NIH grants: 1 F32 DK085835-01A1) (JP), 1 R21 DK081703-01A2 (DC) and 5 P30 DK034854-24 (Harvard Digestive Diseases Center; DC). In addition, we would like to thank the Burrill family for their generous support through the Burrill Research Grant. We would also like to thank members of the Steen Laboratory at Children’s Hospital Boston, in particular John FK Sauld, Ali Ghoulidi, Aleksander Gaun, and Dominic Winter for their technical assistance and critical reading of the manuscript. In addition, we thank members of the Center for Pancreatic Disease at Brigham and Women’s Hospital, particularly Jessica Rosenblum for her technical assistance.
Funding Information:
Guarantor of the article: Joao A. Paulo, PhD. Specific author contributions: J.P. and V.K. carried out the experiments and drafted the original manuscript. J.P., J.B., H.S., and D.C. conceived of the study, and participated in its design and coordination. All authors helped to draft the manuscript and approved the final manuscript. Financial support: Funds were provided by the following NIH grants: 1 F32 DK085835-01A2 (J.P.), 1 R21 DK081703-01A2 (D.C.) and 5 P30 DK034854-24 (Harvard Digestive Diseases Center; D.C.). Potential competing interests: None.
ASJC Scopus subject areas
- Gastroenterology