{
  "abstract": "Background Mucosal melanomas (MM) arise from mucosal melanocytes at various anatomical sites. These tumors are rare, highly aggressive, and often associated with poor outcomes. Current treatments, including immune checkpoint inhibitors, show limited efficacy in advanced disease. Compared with cutaneous melanomas, there is a lack of data on the immunogenicity and interferon (IFN)-γ sensitivity of MM. In this study, we examined these features in sino-nasal melanomas (SN-MM) cell lines and clinical samples using microscopy and functional genomics.Methods The immune contexture of SN-MM was analyzed by immunohistochemistry on 48 tumor biopsies. RNA sequencing and mass spectrometry-based proteomic approaches were used to study the IFN-γ receptor (IFNGR) pathways in five patient-derived SN-MM cell lines. Moreover, their IFN-γ sensitivity, in terms of cell viability, IFNGR/JAK/STAT signaling pathway and IFN-γ inducible proteins, was evaluated by flow cytometry and immunoblots. Neoantigen prediction was performed through integrated whole exome sequencing and RNA-sequencing analysis using pVAC-Seq. Immune effector functions were evaluated in co-culture in vitro assays.Results SN-MM tumors are mainly immune “desert” with few tumor-infiltrating lymphocytes and contain immunosuppressive macrophages, features linked to poor prognosis; moreover, tumor cells are largely CD274/programmed death-ligand 1 negative. SN-MM cell lines express transcripts for melanocytic and cancer testis antigens; moreover, sequencing analysis identified a repertoire of high-confidence neoantigens, including candidates derived from recurrently mutated oncogenic drivers. Functional assays revealed that SN-MM cells are susceptible to NK cell-mediated killing. In terms of IFN-γ sensitivity, SN-MM cells show normal surface expression of IFNGR and maintain the integrity of the IFNGR/JAK/STAT signaling pathway. Transcriptomic and proteomic analyses demonstrate that SN-MM cell lines, as a group, respond to IFN-γ by upregulating genes involved in immune recognition and antigen presentation. In 60% of SN-MM lines, IFN-γ also induces cytotoxic and anti-proliferative effects, the release of CXCL10 and upregulation of CD274/PD-L1. The remaining SN-MM cell lines, characterized by poor differentiation, show refractoriness to these effects.Conclusions SN-MM displays an immune-desert phenotype yet retains intrinsic immunogenicity. Most tumors preserve functional IFN-γ signaling, while poorly differentiated cells show resistance to IFN-γ-mediated effects. These findings underscore heterogeneity in immune responsiveness and support functional immune profiling to refine immunotherapy strategies in MM.",
  "authors": [
    {
      "affiliations": [
        "Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy"
      ],
      "name": "Matilde Monti"
    },
    {
      "affiliations": [
        "Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy"
      ],
      "name": "Sara Picinoli"
    },
    {
      "affiliations": [
        "Unit of Pathology, ASST Spedali Civili di Brescia, Brescia, Italy"
      ],
      "name": "Anna Bozzola"
    },
    {
      "affiliations": [
        "Department of Neuroscience, University of Padua, Padua, Italy",
        "Unit of Otorhinolaryngology – Head and Neck Surgery, Azienda Ospedale Universita Padova, Padua, Italy"
      ],
      "name": "Marco Ferrari"
    },
    {
      "affiliations": [
        "Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy",
        "Unit of Pathology, ASST Spedali Civili di Brescia, Brescia, Italy"
      ],
      "name": "Mattia Bugatti"
    },
    {
      "affiliations": [
        "Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy"
      ],
      "name": "Irene Pezzali"
    },
    {
      "affiliations": [
        "Department of Biotechnology, University of Verona, Verona, Italy"
      ],
      "name": "Marco Carlomagno"
    },
    {
      "affiliations": [
        "Department of Biotechnology, University of Verona, Verona, Italy"
      ],
      "name": "Giada Carta"
    },
    {
      "affiliations": [
        "Department of Biotechnology, University of Verona, Verona, Italy"
      ],
      "name": "Matteo Orlandi"
    },
    {
      "affiliations": [
        "Department of Biotechnology, University of Verona, Verona, Italy"
      ],
      "name": "Giacomo Lora"
    },
    {
      "affiliations": [
        "Department of Transfusion Medicine, ASST Spedali Civili di Brescia, Brescia, Italy"
      ],
      "name": "Luisa Benerini Gatta"
    },
    {
      "affiliations": [
        "Department of Otorhinolaryngology, Head and Neck Surgery, Maastricht University Medical Centre+, Maastricht, The Netherlands"
      ],
      "name": "Francesco Missale"
    },
    {
      "affiliations": [
        "Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy"
      ],
      "name": "Giorgia Ferrari"
    },
    {
      "affiliations": [
        "Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy"
      ],
      "name": "Valentina Baldazzi"
    },
    {
      "affiliations": [
        "Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy"
      ],
      "name": "Sara Rezzola"
    },
    {
      "affiliations": [
        "Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy"
      ],
      "name": "Giovanna Tabellini"
    },
    {
      "affiliations": [
        "Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy",
        "National Center for Gene Therapy and Drugs based on RNA Technology - CN3, Padova, Italy",
        "Consorzio Interuniversitario per le Biotecnologie (CIB), Trieste, Italy"
      ],
      "name": "Silvia Parolini"
    },
    {
      "affiliations": [
        "Department of Translational Medicine, University of Eastern Piedmont Amedeo Avogadro School of Medicine, Novara, Italy",
        "Center for Autoimmune and Allergic Diseases, University of Eastern Piedmont Amedeo Avogadro School of Medicine, Novara, Italy"
      ],
      "name": "Marcello Manfredi"
    },
    {
      "affiliations": [
        "Department of Translational Medicine, University of Eastern Piedmont Amedeo Avogadro School of Medicine, Novara, Italy",
        "Center for Autoimmune and Allergic Diseases, University of Eastern Piedmont Amedeo Avogadro School of Medicine, Novara, Italy"
      ],
      "name": "Veronica De Giorgis"
    },
    {
      "affiliations": [
        "Department of Sciences and Technological Innovation, University of Eastern Piedmont Amedeo Avogadro - Alessandria Campus, Alessandria, Italy"
      ],
      "name": "Emilio Marengo"
    },
    {
      "affiliations": [
        "Unit of Otorhinolaryngology and Head and Neck Surgery, University of Insubria Department of Biotechnology and Life Sciences, Varese, Italy"
      ],
      "name": "Mario Turri-Zanoni"
    },
    {
      "affiliations": [
        "Department of Neuroscience, University of Padua, Padua, Italy",
        "Unit of Otorhinolaryngology – Head and Neck Surgery, Azienda Ospedale Universita Padova, Padua, Italy"
      ],
      "name": "Piero Nicolai"
    },
    {
      "affiliations": [
        "Unit of Oncology, ASST Spedali Civili di Brescia, Brescia, Italy"
      ],
      "name": "Francesca Consoli"
    },
    {
      "affiliations": [
        "Department of Medical and Surgical Specialties, Università degli Studi di Brescia, Brescia, Italy"
      ],
      "name": "Davide Lombardi"
    },
    {
      "affiliations": [
        "Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy"
      ],
      "name": "Paolo Martini"
    },
    {
      "affiliations": [
        "Medicine, Washington University School of Medicine in Saint Louis, St. Louis, Missouri, USA"
      ],
      "name": "Jinglun Li"
    },
    {
      "affiliations": [
        "Washington University in St Louis School of Medicine, St Louis, Missouri, USA"
      ],
      "name": "Obi Griffith"
    },
    {
      "affiliations": [
        "Washington University in St Louis School of Medicine, St Louis, Missouri, USA"
      ],
      "name": "Malachi Griffith"
    },
    {
      "affiliations": [
        "Department of Biotechnology, University of Verona, Verona, Italy"
      ],
      "name": "Marzia Rossato"
    },
    {
      "affiliations": [
        "Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy",
        "Pathology and Immunology, Washington University School of Medicine in Saint Louis, St. Louis, Missouri, USA"
      ],
      "name": "William Vermi"
    }
  ],
  "title": "Integrating interferon gamma receptor pathways, antigenicity, and immune contexture as predictors of immunotherapeutic strategies for mucosal melanomas",
  "uid": "c6cc29e4-bafb-51f2-8703-1aabdad15e2d"
}