Analysis of colon tissue sample from Korean patients with CD(Crohn’s Disease)using liquid chromatography and tandem mass spectrometry : 액체크로마토그래피와 직렬 질량분석기를 이용한 한국인 크론병 환자 샘플의 분석

다국어 초록 (Multilingual Abstract)

Crohn’s disease (CD) is a major form of inflammatory bowel disease (IBD). Recently, diagnosis of IBD is very difficult to correctly diagnose. Although there are several diagnostic criteria, there is no single gold standard marker. Also, no definitive diagnostic test exists for CD. Primary purpose of this research is finding biomarker candidates for diagnosis of CD from Korean CD patient's tissue sample. Therefore if we found several proteins which already found in other studies or found first in our study, we can make diagnostic protein chip throughout combination of already-known proteins and novel proteins. Here, we performed the protein expression of Korean CD patient’s tissue samples using label free quantification, especially Corra v.3.1, a computational framework and tools for discovery-based LC-MS/MS proteomics. The comparison between tissue samples of three groups revealed a lot of proteins with at least two-fold changes in their expression level. The protein profiles of colonic epithelium were compared with (i) normal colon, (ii) activated colonic tissue from CD patient, (iii) inactivated colonic tissue from CD patient. Considering the biomarker from a different point-view, it is the protein expressed in patient's guts differently from normal guts. Comparing between protein expression of active CD and that of inactive CD may clue of pathology of disease. Six novel proteins which have higher fold change than 2 in up-regulated in activated CD compared to normal sample are found: PRG2, ATP51, LCP1, PSME1, HNRNPM (isoform 1 of heterogeneous nuclear rib-nucleoprotein M), and PFN1. These proteins are biomarker candidate for diagnosis Crohn’s disease and new therapeutic target of Crohn’s disease.

목차 (Table of Contents)

• I. INTRODUCTION
• 1. Epidemiology
• 2. Clinical menifestation
• A. Ulcerative colitis
• B. Crohn's disease
• 3. Etiology and pathogenesis
• A. Genetics
• B. Immunobiology
• 4. Diagnostic methods up to recently
• 5. Biomarkers identified up to recently
• A. Serological markers
• B. Fecal markers
• 6. Difference between IBD in Korea and IBD in Western conturies
• 7. Introduction of proteomics techniques used in the experiment
• A. Indroduction of proteomics
• B. Mass spectrometry based proteomics
• C. History of mass spectrometry
• D. High-throughput work (properties of MS data) E. Proteomic bioinformatics
• II. PURPOSE
• III. MATERIALS AND METHODS
• 1. Work flow
• 2. Patients
• 3. Reagent
• 4. Protein extraction
• 5. Tryptic digestion
• 6. Liquid chromatography mass spectrometry analysis
• 7. Software tool
• IV. RESULTS
• 1. Total proteins to be identifeid
• 2. Corra data
• 3. The list of proteins that are up- and down- regulated in each group compared to another group
• V. DISCUSSION
• 1. Up-regulated proteins in Activated CD compared to normal sample.
• 2. Down-regulated proteins in Activated CD compared to normal samples.
• 3. Up-regulated proteins in Activated CD compared to Inactivated CD.
• 4. Down-regulated proteins in Activated CD compared to Inactivated CD.
• VI. CONCLUSION AND LIMITATION