Metabolites Profiling of Melanoma Interstitial Fluids Reveals Uridine Diphosphate as Potent Immune Modulator Capable of Limiting Tumor Growth

Eleonora Vecchio; Carmen Caiazza; Selena Mimmi; Angelica Avagliano; Enrico Iaccino; Teresa Brusco; Nancy Nisticò; Domenico Maisano; Annamaria Aloisio; Ileana Quinto; Maurizio Renna; Giuseppina Divisato; Simona Romano; Martina Tufano; Massimo D'Agostino; Elena Vigliar; Antonino Iaccarino; Chiara Mignogna; Francesco Andreozzi; Gaia Chiara Mannino; Rosangela Spiga; Mariano Stornaiuolo; Alessandro Arcucci; Massimo Mallardo; Giuseppe Fiume

doi:10.3389/fcell.2021.730726

Metabolites Profiling of Melanoma Interstitial Fluids Reveals Uridine Diphosphate as Potent Immune Modulator Capable of Limiting Tumor Growth

Front Cell Dev Biol. 2021 Sep 17:9:730726. doi: 10.3389/fcell.2021.730726. eCollection 2021.

Authors

Eleonora Vecchio¹, Carmen Caiazza², Selena Mimmi¹, Angelica Avagliano³, Enrico Iaccino¹, Teresa Brusco², Nancy Nisticò¹, Domenico Maisano¹, Annamaria Aloisio¹, Ileana Quinto¹, Maurizio Renna², Giuseppina Divisato², Simona Romano², Martina Tufano², Massimo D'Agostino², Elena Vigliar³, Antonino Iaccarino³, Chiara Mignogna⁴, Francesco Andreozzi⁵, Gaia Chiara Mannino⁵, Rosangela Spiga⁵, Mariano Stornaiuolo⁶, Alessandro Arcucci³, Massimo Mallardo², Giuseppe Fiume¹

Affiliations

¹ Department of Experimental and Clinical Medicine, University of Catanzaro "Magna Graecia", Catanzaro, Italy.
² Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
³ Department of Public Health, University of Naples Federico II, Naples, Italy.
⁴ Department of Health Sciences, Magna Graecia University, Catanzaro, Italy.
⁵ Department of Medical and Surgical Sciences, University "Magna Graecia" of Catanzaro, Catanzaro, Italy.
⁶ Department of Pharmacy, University of Napoli Federico II, Naples, Italy.

Abstract

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 CD4⁺CD25⁺FoxP3^- 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^hi tumor-associated macrophage (TAM) and of CD3⁺CD8⁺ and CD3⁺CD4⁺ 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.

Keywords: B16 engraftment; TIL (tumor infiltrating lymphocytes); UDP; anti-tumor immunity; metabolite profiling.

Copyright © 2021 Vecchio, Caiazza, Mimmi, Avagliano, Iaccino, Brusco, Nisticò, Maisano, Aloisio, Quinto, Renna, Divisato, Romano, Tufano, D’Agostino, Vigliar, Iaccarino, Mignogna, Andreozzi, Mannino, Spiga, Stornaiuolo, Arcucci, Mallardo and Fiume.