Reactive oxygen species reprogram macrophages to suppress antitumor immune response through the exosomal miR-155-5p/PD-L1 pathway.
Reactive oxygen species reprogram macrophages to suppress antitumor immune response through the exosomal miR-155-5p/PD-L1 pathway.
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DOI：
10.1186/s13046-022-02244-1
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发表时间：
2022-01-27
期刊：
影响因子：
--
通讯作者：
He J
中科院分区：
文献类型：
--
作者：
Li X;Wang S;Mu W;Barry J;Han A;Carpenter RL;Jiang BH;Peiper SC;Mahoney MG;Aplin AE;Ren H;He J
癌细胞的氧化还原(氧化还原)动态平衡失衡。对氧化还原微环境改变对肿瘤免疫反应的确切机制和影响的了解是有限的。我们通过超速离心法从卵巢癌细胞中分离出外切体，并用Western-blotts和纳米颗粒示踪分析对其进行了表征。采用二维、三维共培养肿瘤模型和三维活体细胞成像技术研究肿瘤细胞、巨噬细胞和CD3T细胞之间的相互作用。在裸鼠移植模型和免疫活性小鼠模型中评价外体miR-155-5p在肿瘤生长中的作用。采用流式细胞术和流式细胞术检测腹水和脾巨噬细胞中miR-155-5p和PD-L1的表达水平及CD3T细胞亚群的百分比。活性氧簇(ROS)的升高大大下调了肿瘤细胞外体miR-155-5p的表达。用N-乙酰-L-半胱氨酸(NAC)中和ROS后，巨噬细胞摄取的肿瘤外体miR-155-5p水平增加，导致巨噬细胞迁移减少，肿瘤球体浸润减少。我们进一步发现，程序性死亡配体1(PD-L1)是miR-155-5p的一个功能靶点。经NAC来源的肿瘤外切体或外切体miR-155-5p处理的巨噬细胞与T淋巴细胞共培养后，CD8+T淋巴细胞比例增加，而CD3+T细胞的凋亡率通过PD-L1下调而减少。NAC来源的肿瘤外切体能延缓裸鼠的肿瘤生长。将外源miR-155-5p导入免疫未受损的小鼠体内，可抑制卵巢癌的进展和巨噬细胞的侵袭，并激活CD8+T细胞功能。值得注意的是，exo-miR-155-5p比PD-L1抗体更有效地抑制肿瘤生长，这表明除了PD-L1外，其他途径也可能是该方法的靶点。我们的发现证明了一种新的机制，即ROS诱导miR-155-5p的下调，通过这种机制，肿瘤调节有利于肿瘤生长的微环境。了解ROS对肿瘤免疫反应的负面影响将改进目前的治疗策略。靶向miR-155-5p可以通过下调PD-L1和其他免疫抑制因子来预防免疫抑制TME的形成。网上版载有补充材料，可在10.1186/s13046-022-02244-1查阅。
Cancer cells have an imbalance in oxidation-reduction (redox) homeostasis. Understanding the precise mechanisms and the impact of the altered redox microenvironment on the immunologic reaction to tumors is limited. We isolated exosomes from ovarian cancer cells through ultracentrifuge and characterized by Western-blots and Nanoparticle Tracking Analysis. 2D, 3D-coculture tumor model, and 3D live cell imaging were used to study the interactions between tumor cells, macrophages and CD3 T cells in vitro. The role of exosomal miR-155-5p in tumor growth was evaluated in xenograft nude mice models and immune-competent mice models. Flow cytometry and flow sorting were used to determine the expression levels of miR-155-5p and PD-L1 in ascites and splenic macrophages, and the percentages of CD3 T cells subpopulations. The elevation of reactive oxygen species (ROS) greatly downregulated exosomal miR-155-5p expression in tumor cells. Neutralization of ROS with N-acetyl-L-cysteine (NAC) increased the levels of miR-155-5p in tumor exosomes that were taken up by macrophages, leading to reduction of macrophage migration and tumor spheroid infiltration. We further found that programmed death ligand 1 (PD-L1) is a functional target of miR-155-5p. Co-culture of macrophages pre-treated with NAC-derived tumor exosomes or exosomal miR-155-5p with T-lymphocytes leading to an increased percentage of CD8+ T-lymphocyte and a decreased CD3+ T cell apoptosis through PD-L1 downregulation. Tumor growth in nude mice was delayed by treatment with NAC-derived tumor exosomes. Delivery of tumor exo-miR-155-5p in immune-intact mice suppressed ovarian cancer progression and macrophage infiltration, and activated CD8+ T cell function. It is of note that exo-miR-155-5p inhibited tumor growth more potently than the PD-L1 antibody, suggesting that in addition to PD-L1, other pathways may also be targeted by this approach. Our findings demonstrate a novel mechanism, ROS-induced down-regulation of miR-155-5p, by which tumors modulate the microenvironment that favors tumor growth. Understanding of the negative impact of ROS on the tumor immune response will improve current therapeutic strategies. Targeting miR-155-5p can be an alternative approach to prevent formation of an immunosuppressive TME through downregulation of PD-L1 and other immunosuppressive factors. The online version contains supplementary material available at 10.1186/s13046-022-02244-1.
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影响因子：
6.6
作者：
Lin, Xuzhu;Zheng, Wei;Shen, Pingping
通讯作者：
Shen, Pingping
影响因子：
10.1
作者：
Martinez-Usatorre, Amaia;Sempere, Lorenzo F.;Romero, Pedro
通讯作者：
Romero, Pedro
外泌体在氧化应激期间传达保护性信息；外泌体穿梭RNA的可能作用。
影响因子：
3.7
作者：
Eldh M;Ekström K;Valadi H;Sjöstrand M;Olsson B;Jernås M;Lötvall J
通讯作者：
Lötvall J
影响因子：
6.4
作者：
Coburn SB;Bray F;Sherman ME;Trabert B
通讯作者：
Trabert B
影响因子：
32.4
作者：
Dudda JC;Salaun B;Ji Y;Palmer DC;Monnot GC;Merck E;Boudousquie C;Utzschneider DT;Escobar TM;Perret R;Muljo SA;Hebeisen M;Rufer N;Zehn D;Donda A;Restifo NP;Held W;Gattinoni L;Romero P
通讯作者：
Romero P