Title Page
Abstract
Contents
1. Introduction 11
1.1. Cause and characteristics of CADASIL 11
1.2. Biomarkers of CADASIL and its application in clinics 13
1.3. Proteomics research of CADASIL 16
2. Materials and Methods 17
2.1. Information of clinical samples 17
2.2. Sample Preparation 22
2.2.1. Plasma samples depletion 22
2.2.2. Peptidization for LC-MS analysis 22
2.3. Nano-LC-ESI-MS/MS Analysis 24
2.4. Protein Identification by Database Search 25
3. Results 26
3.1. Plasma protein expression between CADASIL, Stroke and Healthy groups 26
3.1.1. Commonalities and differences in protein expression among three groups 26
3.1.2. Downregulation of TCOF1 expression in CADASIL patients; however, the expression of SGTA increases. 36
3.2. The CADASIL patient group had distinct differences from other groups. 48
3.3. Discussion 54
References 63
국문요약 67
Table 1. Potential biomarkers detected in the blood of CADASIL provided by Jeju National University 14
Table 2. Potential biomarkers detected in the brain vessels of CADASIL provided by Jeju National University 15
Table 3. CADASIL patient group's patient information provided by Jeju National University 18
Table 4. Age information by group provided by Jeju National University 20
Table 5. 112 proteins specific to the CADASIL patient group shown by Venn diagram 34
Table 6. 7 proteins specific to the stroke patient group shown by Venn diagram 35
Figure 1. Box plot showing the spectrum abundance of each sample by Proteome Discoverer 27
Figure 2. Comparison of proteins from a CADASIL patient group with a healthy control group (A) Venn diagram showing the number of common and specific proteins in each group (B) Volcano plot showing... 29
Figure 3. Comparison of proteins from a CADASIL patient group with a stroke patient group (A) Venn diagram showing the number of common and specific proteins in each group (B) Volcano plot showing... 30
Figure 4. Comparison of proteins from a CADASIL patient group with a stroke patient group and a healthy control group (A) Venn diagram showing the number of common and specific proteins in each... 32
Figure 5. Bar graph showing the relative protein abundance of TCOF1 per sample 37
Figure 6. ROC curves and interactive plots of TCOF1 in CADASIL patient group (A) ROC curve and interactive plot of TCOF1 in CADASIL patient group versus two groups (B) ROC curve and interactive... 38
Figure 7. Bar graph showing the relative protein abundance of Small glutamine-rich tetratricopeptide repeat-containing protein alpha (SGTA) per sample 40
Figure 8. ROC curves and interactive plots of SGTA in CADASIL patient group (A) ROC curves and interactive plots of SGTA in CADASIL patient group versus two groups (B) ROC curves and interactive... 41
Figure 9. ROC curves and interactive plots of TCOF1 and SGTA in CADASIL patient group (A) ROC curves and interactive plots of TCOF1 and SGTA in CADASIL patient group versus two groups (B)... 43
Figure 10. The biological general repository for interaction datasets (BioGRID) analysis suggests potential substrates of TCOF1 (https://thebiogrid.org/ (accessed on 19 April 2023)) (A) The overall... 45
Figure 11. RNA and protein expression of TCOF1 (https://www.proteinatlas.org/ (accessed on 19 April 2023)) (A) RNA and protein expression summary of TCOF1. The bar on the left represents RNA... 46
Figure 12. The biological general repository for interaction datasets (BioGRID) analysis suggests potential substrates of SGTA (https://thebiogrid.org/ (accessed on 19 April 2023)) (A) The overall... 47
Figure 13. Besides TCOF1 and SGTA, some proteins were identified in the CADASIL patient group, while others were only expressed in other groups (A) Proteins that showed increased expression in the... 49
Figure 14. Gene ontology (GO) analysis of the CADASIL patient group and stroke patient group The CADASIL patient group and the stroke patient group were compared with the healthy control group,... 53
Figure 15. Potential biomarkers detected in the blood of CADASIL mentioned in Table 1 and their related proteins were compared with the results of this study 57
Figure 16. Based on the results in Figure 14, each group's proteins showing significant differences are shown (A) Comparison of HtrA Serine Peptidase 1 (HTRA1) expression levels in the three groups (B)... 61