文献标题：Targeting mesothelin-CD24 axis repolarizes tumor-associated macrophages to potentiate PD-1 blockade therapy in high-grade serous ovarian cancer

DOI：10.1136/jitc-2024-011230

发表期刊：Journal for ImmunoTherapy of Cancer

影响因子：10.6

发表时间：2025年2月25日

作者单位：复旦大学附属妇产科医院

引用产品：三标多重免疫荧光试剂盒

高级别浆液性卵巢癌（High-grade serous ovarian cancer, HGSOC）的免疫抑制微环境导致免疫检查点阻断（ICB）疗法疗效有限。该研究揭示间皮素（mesothelin，MSLN）作为关键免疫抑制介质，其高表达与患者不良预后和抗PD-1耐药显著相关。首次阐明MSLN通过激活Wnt/β-catenin信号通路上调CD24表达，促进肿瘤相关巨噬细胞（TAMs）向促肿瘤表型极化，并抑制CD8+ T细胞功能。另外该研究提出靶向MSLN联合抗PD-1治疗可逆转TAMs极化、增强T细胞浸润，并显著抑制肿瘤生长和延长生存期，为HGSOC的精准免疫治疗提供了新策略。

本研究中使用攸碧艾三标多重免疫荧光试剂盒（货号：YB006）进行多重免疫荧光分析，精准可视化肿瘤微环境中癌细胞、TAMs和T细胞的空间分布与表型。该试剂盒的高分辨率多重标记能力为MSLN-CD24轴的机制研究提供了关键空间生物学证据。

Figure 1.MSLN exhibits an inverse correlation with response to anti-PD-1 treatment in HGSOC organoids. (A) Heatmap clustering based on the relative abundance of cleaved caspase-3+ CD45− cells, multiple subsets of CD8+ T cells, TH1 cytokines (IL-2, IFN-γ) and TH1 chemokines (CXCL9, CXCL10) demonstrated anti-PD-1 responses in HGSOC organoids via flow cytometry (n=28). Color codes represent response subgroups, MSLN subgroups, and Z-score (log2 intensity). Complete linkage and Euclidean distance were used for clustering. (B) MSLN expression levels, as indicated by the H-score, in anti-PD-1 responders (n=6) and non-responders (n=22) from HGSOC organoids. (C–D) Proportion of Ki-67 (C) and TNF-α, IFN-γ, TNF-α/IFN-γ, granzyme B/IFN-γ expressing CD8+ T cells (D) in HGSOC organoids stratified by the median H-score of MSLN with isotype or anti-PD-1 treatment (n=28). (E) Heatmap clustering based on the relative abundance of cleaved caspase-3+ CD45− cells and multiple subsets of CD8+ T cells illustrating the response to the combination of anti-PD-1 and LMB-100 versus anti-PD-1 alone in HGSOC organoids, as determined by flow cytometry (n=22). Color codes represent response subgroups, MSLN subgroups, and Z-score (log2 intensity). Complete linkage and Euclidean distance were used for clustering. (F) The proportion of CD8+ and CD68+ cells among CD45+ cells in HGSOC organoids between responders (n=12) and non-responders (n=10) to the combination of anti-PD-1 and LMB-100 treatment. (G–H) Representative immunofluorescence images (G) and contour maps (H) illustrating CD68, CD8a, and Pan keratin staining in responders and non-responders to the combination of anti-PD-1 and LMB-100 treatment. (I) Quantification of the distance from CD8+ T cells (left) and CD68+ macrophages (right) to the nearest Pan keratin+ cell in responders (n=12) and non-responders (n=10) to anti-PD-1+LMB-100 treatment. (J) Proportion of arginase-1, CD206, HLA-DR, CD86, CXCL9 expressing CD68+ macrophages in HGSOC organoids between responders (n=12) and non-responders (n=10) to anti-PD-1+LMB-100 treatment. (K–L) Proportion of TNF-α, IFN-γ, granzyme B, TNF-α/IFN-γ, TNF-α/granzyme B, and IFN-γ/granzyme B expressing CD8+ T cells (K) and arginase-1, CD206, HLA-DR, CD86, CXCL9 expressing CD68+ macrophages (L) in HGSOC organoids under the treatment with isotype IgG, anti-PD-1, LMB-100 alone, or the combination of anti-PD-1 and LMB-100 for 48 hours (n=12), as assessed by flow cytometry. Data are presented as mean±SD. Statistical significance was determined using a two-tailed unpaired Student’s t-test or Mann-Whitney U test (B, F, I–J), two-sided paired Student’s t-test (C–D), and two-way analysis of variance, followed by Tukey’s post hoc test for multiple comparisons (K–L). Con, control; CXCL9, C-X-C motif chemokine ligand 9; CXCL10, C-X-C motif chemokine ligand 10; HGSOC, high-grade serous ovarian cancer; IFN-γ, interferon-γ; IL-2, interleukin-2; MSLN, mesothelin; NS, no significant difference; Pan-CK, pan cytokeratin; PD-1, programmed cell death protein-1; TNF-α, tumor necrosis factor-α.