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研究生： 黃晟嘉
Huang, Cheng-Chia
論文名稱： M2型巨噬細胞來源之可溶性TREM2促進免疫抑制性正向回饋並與口腔鱗狀細胞癌之不良預後相關
M2 Macrophage-Derived Soluble TREM2 Promotes an Immunosuppressive Feedback Loop and Predicts Poor Prognosis in Oral Squamous Cell Carcinoma
指導教授： 謝達斌
Shieh, Dar-Bin
學位類別： 碩士
Master
系所名稱： 醫學院 - 口腔醫學研究所
Institute of Oral Medicine
論文出版年： 2026
畢業學年度： 114
語文別： 英文
論文頁數： 81
中文關鍵詞： 口腔癌 、TREM2 、sTREM2 、免疫 、治療
外文關鍵詞： OSCC, TREM2, sTREM2, macrophage, nanobody
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    • 口腔鱗狀細胞癌（OSCC）為最常見之口腔癌。在台灣與嚼食檳榔、飲酒及吸菸等生活型態密切相關。儘管手術切除、放射治療及化學治療持續進步，OSCC患者五年存活率一直維持在約50%，晚期或轉移性患者預後尤其不理想。尋找新生物標記與治療標的為其重要研究課題。TREM2隸屬於免疫球蛋白超家族之跨膜受體，主要表現於小膠質細胞及巨噬細胞。過去研究顯示其參與多種癌症之抗腫瘤免疫調控，但其在OSCC與其腫瘤微環境（TME）的角色仍不明。TREM2可經ADAM10/ADAM17切割產生可溶型TREM2（sTREM2），並已知與阿茲海默症免疫調控相關。但其對癌症腫瘤微環境巨噬細胞極化之角色尚不清楚。巨噬細胞具高度可塑性，可極化為促發炎之M1型或抗發炎之M2型。兩者對腫瘤進展作用相反：M1促進抗腫瘤免疫，M2則促進腫瘤生長、轉移、血管新生及免疫抑制。然而sTREM2對巨噬細胞極化之影響目前仍不清楚。本研究探討TREM2/sTREM2於OSCC檢體中之臨床病理意義，並分析sTREM2於體外對巨噬細胞極化之影響。結果顯示，OSCC患者血漿sTREM2濃度顯著上升，且與腫瘤晚期及存活率不佳相關；TREM2陽性巨噬細胞亦於晚期腫瘤組織中顯著增加。於所測試之各細胞類型中，M2之TREM2表現量最高，並為胞外sTREM2之主要來源。M2條件培養液及重組sTREM2均能以濃度依賴方式促進巨噬細胞朝M2型極化。上述結果顯示，M2來源之sTREM2可能於OSCC腫瘤微環境中形成自我強化之免疫抑制迴路，並與疾病進展及不良預後相關，顯示其作為生物標記與治療標的潛力。

    Oral Squamous Cell Carcinoma (OSCC) is the most common oral cancer and remains associated with poor prognosis despite advances in surgery, radiotherapy, and chemotherapy. TREM2, an immunoglobulin superfamily receptor expressed in microglia and macrophages, has been implicated in tumor progression and immune regulation in several cancers, but its role in OSCC and the tumor microenvironment (TME) remains unclear. Soluble TREM2 (sTREM2), generated by ADAM10/ADAM17 cleavage, regulates immune responses in neurodegenerative diseases, yet its role in macrophage polarization within cancer TME is unclear. Macrophages polarize into pro-inflammatory M1 or anti-inflammatory M2 phenotypes with opposing effects on tumor progression: M1 macrophages promote anti-tumor immunity, whereas M2 macrophages facilitate tumor growth, metastasis, angiogenesis, and immune suppression. The effects of sTREM2 on macrophage polarization remain largely unknown. This study investigated the clinicopathological significance of TREM2/sTREM2 in OSCC and evaluated the effects of sTREM2 on macrophage polarization in vitro. Plasma sTREM2 was significantly elevated in patients with OSCC and associated with advanced stage and poor overall survival, and TREM2-positive macrophages were enriched in advanced-stage tumor tissues. Among all cell types examined, M2 macrophages showed the highest TREM2 expression and were the predominant source of extracellular TREM2. Both M2-conditioned medium and recombinant sTREM2 promoted M2-like polarization in a concentration-dependent manner, whereas neutralization of extracellular sTREM2 with an anti-TREM2 nanobody effectively attenuated sTREM2-induced M2-like polarization. These findings indicate that M2-derived sTREM2 sustains a self-reinforcing immunosuppressive loop in the OSCC tumor microenvironment and is associated with disease progression and poor prognosis, supporting its potential as a biomarker and therapeutic target in OSCC.

    Abstract 3 中文摘要 4 Acknowledgement 5 Contents 6 1. Introduction 9 1-1 Oral cancer 9 1-2 Triggering receptor expressed on myeloid cells 2 (TREM2) 9 1-3 Soluble form of TREM2 (sTREM2) : structure, function and mechanism 11 1-4 Macrophages : M1 and M2 phenotypes 12 1-5 Rationale 13 2. Materials & Methods 15 2-1 Institutional Review Board (IRB) approval 15 2-2 Measurement of plasma sTREM2 levels by ELISA 15 2-3 Immunohistochemistry (IHC) double staining analysis 16 2-4 TissueFAXS analysis 17 2-5 Tissue Microarray analysis 18 2-6 Cell Culture and Maintenance of hNOK, DOK, THP-1, and Oral Cancer Cell Lines 20 2-7 THP-1 Differentiation and Macrophage Polarization 22 2-8 Tim4-beads exosome isolation protocol 23 2-9 Western Blot analysis 24 2-10 Macrophage Polarization Induced by M2 Macrophage-Conditioned Medium and Recombinant sTREM2 26 2-11 Surface Staining and Cell Fixation for Flow Cytometry 28 2-12 Flow Cytometry analysis 28 3. Results 30 3-1 Elevated plasma sTREM2 levels in patients with OSCC 30 3-2 ROC analysis of plasma sTREM2 for discriminating OSCC patients from healthy donors 30 3-3 Association between plasma sTREM2 levels and clinicopathological characteristics of OSCC 31 3-4 Association between plasma sTREM2 levels and anatomical locations of OSCC lesions 31 3-5 Association of plasma sTREM2 levels with age and sex 32 3-6 Elevated plasma sTREM2 levels are associated with poor overall survival in patients with OSCC 32 3-7 TREM2-positive macrophages are enriched in advanced-stage OSCC tissues 33 3-8 Increased TREM2 expression in OSCC and HNSCC cell lines 34 3-9 Elevated cellular TREM2 expression in M2 macrophages compared with OSCC cell lines 34 3-10 TREM2 is not secreted through exosomes in OSCC cells and macrophages 35 3-11 M2 macrophages predominantly release soluble TREM2 into the culture supernatant 35 3-12 M2 macrophage-derived conditioned medium promotes M2-like macrophage polarization 36 3-13 Recombinant sTREM2 directly induces M2-like macrophage polarization 36 3-14 High concentrations of sTREM2 significantly enhance M2 macrophage polarization 37 3-15 Anti-TREM2 nanobody attenuates sTREM2-induced M2-like macrophage polarization 37 Discussion 39 Summary 47 References 49 Figures 53 Tables 76 Appendix 80

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