Title Page
Abstract
Contents
Chapter 1. Introduction 8
1.1. Glioblastoma 8
1.2. Origin of glioblastoma 8
1.3. Neural stem cells in SVZ as origin of glioblastoma recurrence 10
1.4. Purpose of research 11
Chapter 2. Materials and methods 12
2.1. Mouse modeling 12
2.1.1. Cre-expressing constructs to model p53, Pten mutations 12
2.1.2. Mouse care and information 12
2.1.3. In vivo electroporation 13
2.1.4. Surgical resection 14
2.1.5. Stereotaxic injection of virus into cortex after surgical resection 14
2.1.6. In vivo treatment with AMD3100 15
2.1.7. Sphere culture 15
2.1.8. Orthotopic implantation and surgical removal primary tumors 16
2.2. Immunostaining and histological analysis of mouse brain 16
2.3. MRI imaging of mouse brain 17
2.4. In vitro differentiation analysis treated with AMD3100 17
2.5. RNA sequencing 18
2.5.1. RNA sequencing sampling from mice tissue 18
2.5.2. RNA sequencing sampling from human tissue 18
2.5.3. RNA sequencing analysis 18
2.6. Statistical analysis 19
Chapter 3. Results 21
3.1. Mouse modeling of SVZ-driven glioma at RC after surgical resection 21
3.1.1. Glioma reconstruction at the RC 22
3.1.2. Glioma reconstruction at RC after primary tumor removal 23
3.1.3. Surgical resection and viral transduction of cancer mutation in the cortex 25
3.1.4. Mutant NSCs migrating specifically to RC through OPC lineage 27
3.2. Increased CXCR4 expression at RC after surgical resection 28
3.2.1. RNA sequencing analysis of RC in mice after surgical resection 28
3.2.2. Immunostaining of CXCR4 and CXCL12 expression in mice 29
3.3. Analysis of primary and recurrent tumor tissues in mice and human 30
3.3.1. RNA sequencing analysis of primary and recurrent tumor in mice 30
3.3.2. RNA sequencing analysis of primary and recurrent tumor in human 33
3.3.3. Analysis of CXCR4 expression from public database 37
3.4. Effects of AMD3100 treatment on mutant NSCs in vivo and in vitro 38
Chapter 4. Discussion 41
Chapter 5. Conclusion 45
Reference 46
국문초록 53