{
  "abstract": "Background Innate immune regulators NLRC3, NLRC4 and NLRC5 are emerging as modulators of anti-tumor immunity and may influence immunotherapy sensitivity in melanoma. However, their clinical relevance remains unclear. This study integrates bulk and single-cell transcriptomic analyses to characterize their expression patterns and associations with immune infiltration, checkpoint signaling and clinical outcomes.Methods Transcriptomic and clinical data from 448 melanoma patients were obtained from The Cancer Genome Atlas. We analyzed expression and prognostic impact of NLRC3, NLRC4, and NLRC5. The GSE65904 from Gene Expression Omnibus served as external validation. Correlation between gene expression and survival outcomes was assessed using KM Plotter, which includes 423 immunotherapy-treated melanoma patients. Using CIBERSORT and xCell algorithms, we explored tumor microenvironment (TME) differences between high- and low-expression groups. We analyzed GSE120575, a single-cell RNA (scRNA) sequencing from immunotherapy-treated patients downloaded from TISCH2.0 to identify cell-specific expression of NLRC genes.Results Analysis of the TCGA dataset demonstrated that higher expression levels of NLRC3, NLRC4, and NLRC5 were associated with improved survival outcomes (p<0.05). Additionally, these genes showed a significant positive correlation with the expression of PD-1 and CTLA-4 (p < 0.05). In the validation cohort, NLRC3 and NLRC5 remained significantly associated with better survival (p<0.05), while NLRC4 showed a trend toward improved survival (p=0.096). Consistent findings were observed in immunotherapy-treated cohorts with significantly improved survival outcomes correlating with higher NLRC3 and NLRC5 expression measured by OS and PFS, irrespective of anti-PD-1 or anti-CTLA4 therapy. Notably, NLRC4 was significantly associated with favorable outcomes only in patients treated with anti-CTLA4 (p<0.05). ScRNA analysis revealed elevated expression of NLRC5 and NLRC3 in cytotoxic T cells (CD8+), regulatory T cells (Tregs), CD4+ T cells, proliferating T cells, and natural killer (NK) cell clusters. In contrast, NLRC4 exhibited higher expression in monocyte/macrophages and dendritic cell clusters. TME bulk deconvolution analysis revealed that high expression of NLRC5, NLRC4, and NLRC3 showed significantly higher infiltration levels of CD8+ T cells, memory activated CD4+ T cells, and M1 macrophages. Higher infiltration of M0 macrophages and eosinophils was common in low-expressors (P<0.05). Immune, microenvironment, and stromal scores were elevated in the high-expression groups (p<0.05).Conclusions Higher expression of NLRC gene expression was associated with improved survival and enhanced immune infiltration in melanoma. Their expression predicts response to checkpoint inhibitors. Future studies are needed to validate these findings and apply them in clinical practice. These genes may serve as prognostic biomarkers, identify immunotherapy responders and guide personalized strategies in future trials.",
  "authors": [
    {
      "affiliations": [
        "Southeast Health, Dothan, AL, USA"
      ],
      "name": "Rowan Bandaranaike"
    },
    {
      "affiliations": [
        "Jordan Universities of Science and Technology, Irbid, Jordan"
      ],
      "name": "Mohammad Khaled Alhushki"
    },
    {
      "affiliations": [
        "University of Alabama at Birmingham, Birmingham, AL, USA"
      ],
      "name": "Sanad Alhushki"
    },
    {
      "affiliations": [
        "Jordan Universities of Science and Technology, Irbid, Jordan"
      ],
      "name": "Nour Al-Bzour"
    },
    {
      "affiliations": [
        "University of Alabama at Birmingham, Birmingham, AL, USA"
      ],
      "name": "Sahanna Shiggaon"
    },
    {
      "affiliations": [
        "University of Alabama at Birmingham, Birmingham, AL, USA"
      ],
      "name": "Aakash Desai"
    },
    {
      "affiliations": [
        "University of Alabama at Birmingham, Birmingham, AL, USA"
      ],
      "name": "Maya Khalil"
    }
  ],
  "title": "130 Single-cell and bulk RNA sequencing analysis of NLRC3, NLRC4 and NLRC5 in cutaneous melanoma",
  "uid": "5ec57541-d82d-5374-99b8-73d4b7d59349"
}