Intratumoral combination therapy with poly(I:C) and resiquimod synergistically triggers tumor-associated macrophages for effective systemic antitumoral immunity.
Intratumoral combination therapy with poly(I:C) and resiquimod synergistically triggers tumor-associated macrophages for effective systemic antitumoral immunity.
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DOI：
10.1136/jitc-2021-002408
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发表时间：
2021-09
影响因子：
10.9
通讯作者：
Andón FT
中科院分区：
文献类型：
--
作者：
Anfray C;Mainini F;Digifico E;Maeda A;Sironi M;Erreni M;Anselmo A;Ummarino A;Gandoy S;Expósito F;Redrado M;Serrano D;Calvo A;Martens M;Bravo S;Mantovani A;Allavena P;Andón FT
肿瘤相关巨噬细胞（tam）发挥着关键的免疫抑制作用，限制了免疫系统对抗癌症的能力，并阻碍了目前临床上应用的大多数治疗方法的抗肿瘤效果。先前的研究已经评估了（TLR）激动剂引发的抗肿瘤免疫反应，如poly(I:C)，咪喹莫特（R837）或瑞喹莫特（R848）作为单一疗法；然而，它们的结合治疗癌症还没有被探索过。本研究考察了poly（I:C）与R848或R837联合治疗与单独治疗的抗肿瘤效果和巨噬细胞重编程。采用Alamar Blue、ELISA和流式细胞术对TLR激动剂的体外毒性和免疫刺激活性进行了评估，实验对象为原代人和小鼠m - csf分化的巨噬细胞。用体外功能流式细胞术评价tlr处理的巨噬细胞对癌细胞的细胞毒活性。体内实验采用CMT167肺癌模型和MN/MCA1肺转移纤维肉瘤模型；通过流式细胞术、RT-qPCR、多光谱免疫分型、定量蛋白质组学实验和蛋白相互作用分析评估肿瘤浸润性白细胞。结果表明，poly（I:C）联合R848比单独或联合R837在体外使巨噬细胞向m1样抗肿瘤效应物极化的效果更高。在体内实验中，poly(I:C)+R848在肿瘤内的协同组合可显著阻止肺癌和纤维肉瘤免疫活性小鼠模型的肿瘤生长和转移。退行性肿瘤表现为M1:M2比升高的巨噬细胞浸润增加，CD4+和CD8+ T细胞募集，伴免疫抑制性CD206+ tam和FOXP3+/CD4+ T细胞减少。CD4+和CD8+ T细胞的消耗导致治疗效果完全丧失。通过增强巨噬细胞细胞毒活性和t细胞增殖，治疗小鼠获得了全身抗肿瘤反应和对肿瘤再攻击的抵抗。蛋白质组学实验验证了poly(I:C)+R848组合比单一处理或poly(I:C)+R837更能激活先天免疫，蛋白-蛋白相互作用网络分析揭示了STAT1通路的关键激活。这些发现证实了poly(I:C)+R848联合局部给药后巨噬细胞激活介导的抗肿瘤免疫应答，支持了该疗法在实体瘤患者瘤内的临床应用。
Tumor-associated macrophages (TAMs) play a key immunosuppressive role that limits the ability of the immune system to fight cancer and hinder the antitumoral efficacy of most treatments currently applied in the clinic. Previous studies have evaluated the antitumoral immune response triggered by (TLR) agonists, such as poly(I:C), imiquimod (R837) or resiquimod (R848) as monotherapies; however, their combination for the treatment of cancer has not been explored. This study investigates the antitumoral efficacy and the macrophage reprogramming triggered by poly(I:C) combined with R848 or with R837, versus single treatments. TLR agonist treatments were evaluated in vitro for toxicity and immunostimulatory activity by Alamar Blue, ELISA and flow cytometry using primary human and murine M-CSF-differentiated macrophages. Cytotoxic activity of TLR-treated macrophages toward cancer cells was evaluated with an in vitro functional assay by flow cytometry. For in vivo experiments, the CMT167 lung cancer model and the MN/MCA1 fibrosarcoma model metastasizing to lungs were used; tumor-infiltrating leukocytes were evaluated by flow cytometry, RT-qPCR, multispectral immunophenotyping, quantitative proteomic experiments, and protein–protein interaction analysis. Results demonstrated the higher efficacy of poly(I:C) combined with R848 versus single treatments or combined with R837 to polarize macrophages toward M1-like antitumor effectors in vitro. In vivo, the intratumoral synergistic combination of poly(I:C)+R848 significantly prevented tumor growth and metastasis in lung cancer and fibrosarcoma immunocompetent murine models. Regressing tumors showed increased infiltration of macrophages with a higher M1:M2 ratio, recruitment of CD4+ and CD8+ T cells, accompanied by a reduction of immunosuppressive CD206+ TAMs and FOXP3+/CD4+ T cells. The depletion of both CD4+ and CD8+ T cells resulted in complete loss of treatment efficacy. Treated mice acquired systemic antitumoral response and resistance to tumor rechallenge mediated by boosted macrophage cytotoxic activity and T-cell proliferation. Proteomic experiments validate the superior activation of innate immunity by poly(I:C)+R848 combination versus single treatments or poly(I:C)+R837, and protein–protein-interaction network analysis reveal the key activation of the STAT1 pathway. These findings demonstrate the antitumor immune responses mediated by macrophage activation on local administration of poly(I:C)+R848 combination and support the intratumoral application of this therapy to patients with solid tumors in the clinic.
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DOI：
10.1038/nrclinonc.2016.217
发表时间：
2017-07
期刊：
Nature reviews. Clinical oncology
影响因子：
--
作者：
Mantovani A;Marchesi F;Malesci A;Laghi L;Allavena P
通讯作者：
Allavena P
影响因子：
10.9
作者：
Angela Aznar, M.;Planelles, Lourdes;Melero, Ignacio
通讯作者：
Melero, Ignacio
影响因子：
8
作者：
Huffman, Austin P.;Lin, Jeffrey H.;Vonderheide, Robert H.
通讯作者：
Vonderheide, Robert H.
影响因子：
3.9
作者：
Lu, Ermei;Hu, Xiaoli;Zhu, Xueqiong
通讯作者：
Zhu, Xueqiong
影响因子：
82.9
作者：
Cerezo, Michael;Guemiri, Ramdane;Robert, Caroline
通讯作者：
Robert, Caroline