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Title Page 2

Contents 5

ABSTRACT 10

Ⅰ. Review of literature 14

Ⅱ. Role of the methyltransferase SETD7, a key regulator controlling endothelial-to-mesenchymal transition, in pulmonary arterial hypertension 20

2.1. Introduction 20

2.2. Materials and Methods 26

2.3. Results 32

2.3.1. Expression of methyltransferase SETD7 is upregulated by EndMT-induced PAECs 32

2.3.2. Methyltransferase SETD7 is a key regulator in EndMT 35

2.3.3. Upregulation of SETD7 is associated with EndMT in animal models with pulmonary hypertension 40

2.3.4. SETD7 inhibitor, (R)-PFI-2, recovers SuHx-induced PH by regulating EndMT in rats 44

2.3.5. SETD7 inhibitor, (R)-PFI-2, has an inhibitory effect on EndMT in vitro 49

2.3.6. SETD7 modulates smad2/3 phosphorylation activated by TGF-β2 and IL-1β co-treatment 52

2.3.7. Little evidence of SETD7 acting as an H3K4mel histone methyltransferase in PAECs during EndMT 56

2.3.8. SETD7 regulates protein interaction and downstream pathway of TGF-βR1 through lysine methylation 61

2.4. Discussion 66

Ⅲ. Ginsenoside Rg3 alleviates pulmonary fibrosis by suppressing endothelial-to-mesenchymal transition 70

3.1. Introduction 70

3.2. Materials and Methods 74

3.3. Results 79

3.3.1. Rg3 attenuates BLM-induced PF mice 79

3.3.2. EndMT occurs in the lung of BLM-induced PF mice 81

3.3.3. Rg3 inhibits EndMT in BLM-induced PF 84

3.3.4. Rg3 inhibits EndMT induced by co-treatment with TGF-β2 and IL-1β in HPAECs 87

3.3.5. Rg3 inhibits EndMT by regulating smad signaling 91

3.4. Discussion 93

Ⅳ. Niclosamide formulation attenuates pulmonary vascular remodeling by regulating endothelial-to-mesenchymal transition 98

4.1. Introduction 98

4.2. Materials and Methods 102

4.3. Results 106

4.3.1. Niclosamide formulation ameliorates pulmonary hypertension through vascular muscularization inhibition 106

4.3.2. Niclosamide ameliorates EndMT in PAECs co-treated with TGF-β2 and IL-1β 112

4.3.3. NOTCH2 upregulation is associated with EndMT in animal models with pulmonary hypertension 117

4.3.4. NOTCH2 knockdown ameliorates mesenchymal and PH-related gene expression in PAECs treated with TGF-β2 and IL-1β 119

4.4. Discussion 121

Ⅴ. Summary and Conclusion 125

REFERENCES 129

ABSTRACT IN KOREAN 148

List of Figures 8

Figure 1. Overview of EndMT involved in lung disease 18

Figure 2. SETD7 expression is upregulated in PAECs treated with TGF-β2 and IL-1β 34

Figure 3. Methyltransferase SETD7 plays a key role in EndMT pathogenesis 39

Figure 4. SETD7 is elevated in the endothelium of the SuHx-induced PH rat model 43

Figure 5. SETD7 inhibitors ameliorate pulmonary hypertension through EndMT inhibition 48

Figure 6. (R)-PFI-2 attenuates EndMT and EC dysfunction 51

Figure 7. SETD7 suppresses smad2/3 phosphorylation induced by TGF-β2 and IL-1β 55

Figure 8. SETD7 barely functions as a histone methyltransferase in PAECs treated with TGF-β2 and IL-1β 60

Figure 9. SETD7 methylates TGF-βR1 and regulates TAK1-mediated smad2/3 signaling 65

Figure 10. Rg3 mitigates BLM-induced PF 80

Figure 11. EndMT observed in the lung of BLM-induced PF mice 83

Figure 12. Rg3 restores EndMT in the lung of BLM-induced PF mice 86

Figure 13. Rg3 ameliorates EndMT induced by co-stimulation with TGF-β2 and IL-1β in HPAECs 90

Figure 14. Rg3 suppresses EndMT through smad signaling 92

Figure 15. Niclosamide formulation improves vascular muscularization in lung vessels of SuHx-induced PH rat models 111

Figure 16. Niclosamide inhibits EndMT through NOTCH2 signaling regulation 116

Figure 17. NOTCH2 upregulation during EndMT process in SuHx-induced PH rats 118

Figure 18. NOTCH2 regulates EndMT- and PH-related genes expression 120

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