Vecchio Eleonora, Caiazza Carmen, Mimmi Selena, Avagliano Angelica, Iaccino Enrico, Brusco Teresa, Nisticò Nancy, Maisano Domenico, Aloisio Annamaria, Quinto Ileana, Renna Maurizio, Divisato Giuseppina, Romano Simona, Tufano Martina, D'Agostino Massimo, Vigliar Elena, Iaccarino Antonino, Mignogna Chiara, Andreozzi Francesco, Mannino Gaia Chiara, Spiga Rosangela, Stornaiuolo Mariano, Arcucci Alessandro, Mallardo Massimo, Fiume Giuseppe
Department of Experimental and Clinical Medicine, University of Catanzaro "Magna Graecia", Catanzaro, Italy.
Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
Front Cell Dev Biol. 2021 Sep 17;9:730726. doi: 10.3389/fcell.2021.730726. eCollection 2021.
Tumor interstitial fluid (TIF) surrounds and perfuses tumors and collects ions, metabolites, proteins, and extracellular vesicles secreted by tumor and stromal cells. Specific metabolites, accumulated within the TIF, could induce metabolic alterations of immune cells and shape the tumor microenvironment. We deployed a metabolomic approach to analyze the composition of melanoma TIF and compared it to the plasma of C57BL6 mice, engrafted or not with B16-melanoma cells. Among the classes of metabolites analyzed, monophosphate and diphosphate nucleotides resulted enriched in TIF compared to plasma samples. The analysis of the effects exerted by guanosine diphosphate (GDP) and uridine diphosphate (UDP) on immune response revealed that GDP and UDP increased the percentage of CD4CD25FoxP3 and, on isolated CD4 T-cells, induced the phosphorylation of ERK, STAT1, and STAT3; increased the activity of NF-κB subunits p65, p50, RelB, and p52; increased the expression of Th1/Th17 markers including IFNγ, IL17, T-bet, and RORγt; and reduced the expression of IL13, a Th2 marker. Finally, we observed that local administrations of UDP in B16-engrafted C57BL6 mice reduced tumor growth and necrotic areas. In addition, UDP-treated tumors showed a higher presence of MHCII tumor-associated macrophage (TAM) and of CD3CD8 and CD3CD4 tumor-infiltrating T-lymphocytes (TILs), both markers of anti-tumor immune response. Consistent with this, intra-tumoral gene expression analysis revealed in UDP-treated tumors an increase in the expression of genes functionally linked to anti-tumor immune response. Our analysis revealed an important metabolite acting as mediator of immune response, which could potentially represent an additional tool to be used as an adjuvant in cancer immunotherapy.
肿瘤间质液（TIF）包围并灌注肿瘤，收集肿瘤细胞和基质细胞分泌的离子、代谢物、蛋白质及细胞外囊泡。TIF中积累的特定代谢物可诱导免疫细胞的代谢改变并塑造肿瘤微环境。我们采用代谢组学方法分析黑色素瘤TIF的成分，并将其与接种或未接种B16黑色素瘤细胞的C57BL6小鼠的血浆进行比较。在所分析的代谢物类别中，与血浆样本相比，单磷酸和二磷酸核苷酸在TIF中含量丰富。对鸟苷二磷酸（GDP）和尿苷二磷酸（UDP）对免疫反应的影响分析表明，GDP和UDP增加了CD4CD25FoxP3的百分比，并且在分离的CD4 T细胞上，诱导了细胞外信号调节激酶（ERK）、信号转导和转录激活因子1（STAT1）及信号转导和转录激活因子3（STAT3）的磷酸化；增加了核因子κB（NF-κB）亚基p65、p50、RelB和p52的活性；增加了包括γ干扰素（IFNγ）、白细胞介素17（IL17）、T盒转录因子（T-bet）和维甲酸相关孤儿受体γt（RORγt）在内的辅助性T细胞1（Th1）/辅助性T细胞17（Th17）标志物的表达；并降低了辅助性T细胞2（Th2）标志物白细胞介素13（IL13）的表达。最后，我们观察到在接种B16的C57BL6小鼠中局部给予UDP可减少肿瘤生长和坏死区域。此外，经UDP处理的肿瘤显示出更高比例的主要组织相容性复合体II类（MHCII）肿瘤相关巨噬细胞（TAM）以及CD3CD8和CD3CD4肿瘤浸润性T淋巴细胞（TIL），这两种都是抗肿瘤免疫反应的标志物。与此一致的是，肿瘤内基因表达分析显示，经UDP处理的肿瘤中与抗肿瘤免疫反应功能相关的基因表达增加。我们的分析揭示了一种作为免疫反应介质的重要代谢物，它可能代表了一种可作为癌症免疫治疗佐剂的额外工具。
