Title Page

Contents

Abstract 11

Chapter 1. Background 13

Chapter 2. Investigating the potential effect of adding statins to TKI therapy on molecular response rates in CML treatment 15

2.1. Introduction 15

2.2. Materials and Methods 18

2.2.1. Analysis of DMR rates in patients with CML who were treated with imatinib alone or in combination with a statin 18

2.2.2. Propensity score matching (PSM) analysis 20

2.2.3. Statistical analysis 21

2.2.4. Evaluation of in vitro cytotoxicity against CML cell lines 21

2.2.5. Determining the phosphor-CrkL/CrkL ratio 25

2.2.6. Synergy calculations 25

2.2.7. Colony-formation assay 25

2.2.8. Isolation of hematopoietic progenitor cells from patients with CML/healthy individual 27

2.3. Results 28

2.3.1. Clinical benefits from the use of statins in CML treatment with imatinib therapy 28

2.3.2. Statins synergistically potentiate the cytotoxic activity of TKIs against the BCR-ABL1⁺ cell lines 33

2.3.3. Statins suppress the colony-forming capacity of murine CML-KLS⁺ cells in vitro 42

2.3.4. Combination of rosuvastatin and imatinib exert growth-inhibitory effects against CML CD34⁺ cells 43

2.4. Discussion 46

Chapter 3. Investigating the mechanism underlying the synergistic effect of statin and imatinib combination 48

3.1. Introduction 48

3.2. Materials and Methods 49

3.2.1. RNA sequencing 49

3.2.2. Bioinformatics analysis 49

3.2.3. Gene expression analysis 50

3.2.4. Targeted RNA sequencing assay 51

3.2.5. Pathway enrichment analysis 51

3.3. Results 53

3.3.1. Statins target the c-Myc and hematopoietic stem cell differentiation pathways in CML 53

3.4. Discussion 57

Chapter 4. Conclusion 59

References 61

Appendix 72

논문요약 88

Table 2.1. Drug administration 24

Table 2.2. Demographics and disease characteristics of patients at the time of imatinib therapy initiation 29

Table 3.1. Pathway enrichment analysis of differentially expressed genes between the control and rosuvastatin-treated groups 54

Table 3.2. List of candidate genes that the overlap with those determined in the pathway enrichment analysis using DAVID 56

Figure 2.1. Overall study design and workflow. 17

Figure 2.2. Statins enhanced the rate of deep molecular response in CML patients undergoing imatinib therapy. 32

Figure 2.3. Effect of imatinib and rosuvastatin alone or in combination on the viability of K562/WT, K562/T315Imut, and BaF3/T315Imut cells.[이미지참조] 34

Figure 2.4. Effect of statins and/or TKIs on K562 cell viability. 37

Figure 2.5. Growth-inhibitory effects of the combination of rosuvastatin and tyrosine kinase inhibitors against various BaF3/mutant cells. 39

Figure 2.6. Heatmap and synergy plot of K562/WT cells after rosuvastatin/imatinib treatment. 41

Figure 2.7. Effect of statins on murine CML-KLS⁺ cells and human-derived cells in vitro. 44

Figure 3.1. RNA sequencing analysis reveals that the combination of a statin and tyrosine kinase inhibitor downregulates the c-Myc and hematopoietic stem cell... 55