臺灣博碩士論文加值系統

在 M1/M2 範圍內，CSF-1 (M-CSF) 對於支持巨噬細胞分化、極化和激活有很大的貢獻。有趣的是，腫瘤細胞也能夠為腫瘤微環境產生 M-CSF，並通過促進 M2 樣巨噬細胞極化來支持腫瘤的進展。如今，通過阻斷 M-CSF 信號通路來對抗癌症相關疾病的策略已在臨床研究中廣泛採用，以防止 M2 樣分化。然而，根據一些文獻，阻斷 M-CSF 是自相矛盾的，因為它既可以縮小腫瘤，也可以使腫瘤增大。我們之前的研究表明，雖然 M-CSF 被撤回，但 CSF-1 驅動的骨髓來源的 M0 巨噬細胞 (BMDM) 被引導通過內源性 CSF-1 激活重新編程為 M2 樣狀態。其他人也證明，在小鼠 CSF-1R 的阻斷下，巨噬細胞可以通過 CSF-1 獨立增殖程序指導線蟲感染引起的 Th2 反應的生存能力和擴張。為了進一步了解巨噬細胞中 Th2 反應在 CSF-1 依賴到獨立轉變之間的反應，進行了全面的轉錄組學分析。在 CSF-1 依賴性向 CSF-1 非依賴性轉變後，我們在具有 IL-4 刺激的巨噬細胞中發現了 332 個 DEG。上調的 DEGs 主要涵蓋細胞外基質組織、傷口癒合和脂質衍生物生物合成，而下調的 DEGs 主要代表炎症、免疫反應和趨化因子相互作用。此外，蛋白質-蛋白質相互作用網絡還顯示出 ECM-受體相互作用的上調和炎症相關通路的下調，表明傷口癒合過程得到加強。預計，通過整合我們以前的元數據，可以突出顯示在過渡期間負責 Th2 響應的基因。IL-4 的存在與 CSF-1 非依賴性途徑相結合可以將巨噬細胞的表型改變為載脂巨噬細胞。這是通過其促進脂質衍生物相關活性、促進抗炎反應、免疫耐受和吞噬能力而實現的。

Across the M1/M2 spectrum, CSF-1 (M-CSF) contribute greatly to support macrophages differentiation, polarization, and activation. Intriguingly, tumor cells are also capable of producing M-CSF to the tumor microenvironment and support tumor progression by promoting M2-like macrophages polarization. Nowadays, the strategy to combat cancer-related disease by blocking M-CSF signaling pathways has been adopted widely in the clinical study to prevent M2-like differentiation. However, according to several literatures, poor monotherapy efficacy of anti-CSF1R has been observed and therefore, is not sufficient enough to reduce tumor progression. Our previous study stated that while M-CSF was withdrawn, CSF-1 driven bone marrow derived M0 macrophages (BMDM) were conducted to reprogram into M2-like state via endogenous CSF-1 activation. Others has also demonstrated that under the blockade of CSF-1R in mice, macrophages could direct the survivability and expansion via CSF-1-independent program of proliferation upon Th2 response provoked by nematode infection. To further understand the reaction of Th2 response in macrophages between CSF-1-dependent to independent transition, a comprehensive transcriptomic analysis was proceeded. We found 332 DEGs in macrophages with IL-4 stimulation following CSF-1-dependent to CSF-1-independent transition. The upregulated DEGs mainly covered extracellular matrix organization, wound healing, and lipid-derivatives biosynthesis while the downregulated DEGs mostly represented inflammation, immune response, and chemokine interaction. Furthermore, protein-protein interaction network also displayed an upregulation in ECM-receptor interaction and a downregulation in inflammation-related pathways, suggesting an enhancement in wound healing processes. Expectedly, by integrating our previous metadata, the genes responsible for Th2 response during the transition could be highlighted. The presence of IL-4 in combination with CSF-1-independent pathway could alter the phenotype of macrophages into lipid-laden macrophage, proven by its capability on promoting lipid derivatives-related activity, promoting anti-inflammation response, immune-tolerance, and phagocytosis ability.

摘要 i
ACKNOWLEDEMENT v
TABLE OF CONTENTS vi
LIST OF FIGURES ix
APPENDIX OF CONTENTS xi
I. INTRODUCTION 1
1.1 Background 1
1.2 The Aim of This Study 4
II. LITERATURE REVIEW 5
2.1 Immune System 5
2.1.1 Innate Immune System 6
2.1.2 Adaptive Immune System 7
2.2 Macrophage 8
2.2.1 Lipid-laden Macrophage 10
2.3 Cytokines 11
2.3.1 Th1-type Cytokines 13
2.3.2 Th2-type Cytokines 13

2.4 Colony Stimulating Factor 15
2.4.1 Macrophages-Colony Stimulating Factor 16
2. 5 Transcriptomic Profile 19
III.MATERIALS AND METHODS 22
3.1 Animals 22
3.2 Differentiation of Bone Marrow-Derived Macropahages (BMDMs) 22
3.3 CSF-1 Independent Macrophages Polarization Under IL-4 Stimulation 22
3.4 Library Preparation and RNA-Sequencing 23
3.5 Transcriptome Analysis 24
3.6 Protein-Protein Interaction Analysis 24
3.7 Comprehensive and systematic transcriptomic analysis comparison 24
IV. RESULTS 26
4.1 CSF-1-independent Macrophages Exhibit Higher Capability of Wound Healling and Immune tolerance under IL-4 Stimulation 26
4.2 Protein-Protein Interaction Network Revealed ECM Organization & Remodelling as The Most Significant Network in CSF-1-independent Macrophages Under IL-4 Stimulus 30
4.3 Systematic and Comprehensive Transcriptomic Profiling Analysis Unveiled Distinct Phenotypes in CSF-1-independent Macrophages Before and After IL-4 Administration 32
4.4 Non-overlapped Area Revealed Degs Expressed Exclusively in The Response to The Absence/Presence of IL-4 36
4.5 Overlapped Area Revealed Steroid and Cholesterol Biosynthesis as The Most Unique Phenotypes After Stimulation of IL-4 in The CSF-1-independent Macrophages 39
4.6 Lipid-laden-associated Genes were Upregulated in CSF-1-independent Macrophages After IL-4 Induction 44
V. DISCUSSION 48
VI. CONCLUSION 54
VII. REFERENCES 55
VII. APPENDIX 77

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