IDH2 deficiency inhibits macrophage polarization by disturbed mitochondrial dynamics : IDH2의 결핍 대식세포가 암에 미치는 역할에 관한 연구

국문 초록 (Abstract)

최근 암의 치료적 기법으로 종양 미세환경을 표적으로 암을 치료하는 연구가 활발하게 진행되고있다. 대식세포는 암이 발병되는 종양미세환경에서 특이적으로 분화하여 병증을 악화 또는 완화 시키는 이중적인 기능을 가지고 있다. 본 연구에서 미토콘드리아 시트르산 회로의 한 부분을 담당하고 있는 이소시트르산 탈수소효소 2 (IDH2) 결손이 있는 마우스에서 종양의 성장이 감소하는 원인을 대식세포 분극과 관련성을 두어 알아보고자 한다. 암세포와 공동배양한 IDH2 결핍 대식세포는 미토콘드리아 장애를 유도하여 산화적인산화 반응이 감소하는 것을 확인하였으며, 미토콘드리아 분열을 유도하는 경향을 확인하였다. 또한, IDH2 결핍 대식세포에서 M2 분극화와 관련있는 항염증성 인자가 감소하였으며, M2 대식세포 마커인 Arg1, p-STAT3, p-AMPK도 감소하는 경향을 확인하였다. 우리는 대식세포에서 IDH2의 결핍이 M1와 유사한 대식세포 양극화를 유발하여 암 성장을 억제 할 수 있음을 확인하였다.

다국어 초록 (Multilingual Abstract)

Recently, as a therapeutic technique for cancer, research to treat cancer by targeting the tumor microenvironment is being actively conducted. Macrophages have a dual function of specifically differentiating in the tumor microenvironment where cancer develops, exacerbating or alleviating the disease. The purpose of this study is to investigate the cause of reduced tumor growth in mice with Isocitrate Dehydrogenase 2 (IDH2) deficiency, which is responsible for a part of the mitochondrial TCA cycle, in relation to macrophage polarization. It was confirmed that IDH2-deficient macrophages co-cultured with cancer cells induce mitochondrial dysfunction, thereby reducing oxidative oxidation, and confirming the tendency to induce mitochondrial division. In addition, anti-inflammatory factors related to M2 polarization decreased in IDH2-deficient macrophages, and it was confirmed that the M2 macrophage markers Arg1, p-STAT3, and p-AMPK also tend to decrease. We confirmed that IDH2 deficiency in macrophages could inhibit cancer growth by inducing M1-like macrophage polarization.

목차 (Table of Contents)

• LIST OF FIGURES iii
• Ⅰ. INTRODUCTION 1
• 1. Microenvironment of cancer Tumor associated macrophages 1
• 1.1 Tumor associated macrophages 1
• 1.2 Effect of macrophage polarization on cancer 3
• 2. Isocitrate Dehydrogenase 2 in cancer 7
• 2.1 Mitochondrial function of TCA cycle. 7
• 2.2 Macrophages in mitochondria with IDH2 dysfunction 10
• 2.3 Mitochondrial dynamics dysfunction. 14
• II. MATERIALS AND METHODS 16
• 1.Ragents 16
• 2.Cell culture and Co-culture 16
• 3.Animal preparation 16
• 4.Bone marrow primary culture 17
• 5.Western blot analysis 17
• 6.Real-time PCR assay 19
• 7.Immunohistochemistry 20
• 8.Immunocytochemistry 21
• 9.Determination of mitochondrial ROS levels 21
• 10.Alpha Ketoglutarate quantification assay 22
• 11.NO Detection 23
• 12.Image and measurement of mitochondrial length. 23
• 13.Confocal laser microscopy 24
• 14.Clonogenic assay 24
• 15.Migration assay 25
• 16.Magnetic Activated Cell Sorting (MACS) 25
• 17.Statistical analysis 26
• Ⅲ. RESULTS 27
• 1. IDH2 defect controls polarization of macrophages by mitochondrial dysfunction. 27
• 2. Mitochondrial dynamics in macrophages. 31
• 3. The effect of IDH2 deficiency on macrophage polarization. 34
• 4. The effect of IDH2 deficient macrophage on cancer cells. 39
• 5. The effect of IDH2 deficiency on cancer growth. 43
• IV. DISCUSSION 49
• V. REFERENCES 56
• ABSTRACT (in English) 60
• ABSTRACT (in Korean) 62