Duhamel Marie, Rodet Franck, Delhem Nadira, Vanden Abeele Fabien, Kobeissy Firas, Nataf Serge, Pays Laurent, Desjardins Roxanne, Gagnon Hugo, Wisztorski Maxence, Fournier Isabelle, Day Robert, Salzet Michel
From the ‡Inserm U-1192, Laboratoire de Protéomique, Réponse Inflammatoire, Spectrométrie de Masse (PRISM), Université Lille 1, Cité Scientifique, 59655 Villeneuve D'Ascq, France;
§Institut de Biologie de Lille, UMR 8161 CNRS, Institut Pasteur de Lille, Université Lille 1, Lille, France;
Mol Cell Proteomics. 2015 Nov;14(11):2857-77. doi: 10.1074/mcp.M115.052480. Epub 2015 Sep 1.
Macrophages provide the first line of host immune defense. Their activation triggers the secretion of pro-inflammatory cytokines and chemokines recruiting other immune cells. In cancer, macrophages present an M2 anti-inflammatory phenotype promoting tumor growth. In this way, strategies need to be develop to reactivate macrophages. Previously thought to be expressed only in cells with a neural/neuroendocrine phenotype, the proprotein convertase 1/3 has been shown to also be expressed in macrophages and regulated as a function of the Toll-like receptor immune response. Here, we investigated the intracellular impact of the down-regulation of the proprotein convertase 1/3 in NR8383 macrophages and confirmed the results on macrophages from PC1/3 deficient mice. A complete proteomic study of secretomes and intracellular proteins was undertaken and revealed that inhibition of proprotein convertase 1/3 orient macrophages toward an M1 activated phenotype. This phenotype is characterized by filopodial extensions, Toll-like receptor 4 MyD88-dependent signaling, calcium entry augmentation and the secretion of pro-inflammatory factors. In response to endotoxin/lipopolysaccharide, these intracellular modifications increased, and the secreted factors attracted naïve T helper lymphocytes to promote the cytotoxic response. Importantly, the application of these factors onto breast and ovarian cancer cells resulted in a decrease viability or resistance. Under inhibitory conditions using interleukin 10, PC1/3-knockdown macrophages continued to secrete inflammatory factors. These data indicate that targeted inhibition of proprotein convertase 1/3 could represent a novel type of immune therapy to reactivate intra-tumoral macrophages.
巨噬细胞构成宿主免疫防御的第一道防线。它们的激活会触发促炎细胞因子和趋化因子的分泌，从而招募其他免疫细胞。在癌症中，巨噬细胞呈现出促进肿瘤生长的M2抗炎表型。因此，需要制定策略来重新激活巨噬细胞。前蛋白转化酶1/3以前被认为仅在具有神经/神经内分泌表型的细胞中表达，现已证明它也在巨噬细胞中表达，并作为Toll样受体免疫反应的一个功能受到调节。在此，我们研究了前蛋白转化酶1/3在NR8383巨噬细胞中下调的细胞内影响，并在来自PC1/3缺陷小鼠的巨噬细胞上证实了结果。我们对分泌蛋白组和细胞内蛋白进行了完整的蛋白质组学研究，结果显示，抑制前蛋白转化酶1/3可使巨噬细胞向M1激活表型转变。这种表型的特征是丝状伪足延伸、Toll样受体4 MyD88依赖性信号传导、钙内流增加以及促炎因子的分泌。对内毒素/脂多糖的反应中，这些细胞内修饰增加，分泌的因子吸引幼稚T辅助淋巴细胞以促进细胞毒性反应。重要的是，将这些因子应用于乳腺癌和卵巢癌细胞会导致细胞活力降低或耐药性降低。在使用白细胞介素10的抑制条件下，PC1/3基因敲低的巨噬细胞继续分泌炎症因子。这些数据表明，靶向抑制前蛋白转化酶1/3可能代表一种重新激活肿瘤内巨噬细胞的新型免疫疗法。
