{
  "abstract": "Background Despite recent advances, hepatocellular carcinoma (HCC) remains a leading cause of cancer-related mortality, particularly in advanced stages where curative options are limited. The complement system, a key component of immune response, has emerged as a dual modulator in cancer, capable of promoting or suppressing tumor growth through its effect on the tumor microenvironment (TME). Aberrant complement activation, especially via C3a and C5a signaling, contributes to inflammation, immune evasion, and tumor progression. However, its specific role in HCC remains poorly defined. Elucidating complement dysregulation and interaction may uncover novel prognostic markers and therapeutic targets to improve HCC treatment outcomes.Methods In human HCC tissues, complement mRNA and protein levels were analyzed using nanoString and mass spectrometry respectively. Proteomic findings were validated by immunoblotting and quantitative multiplex immunofluorescence (qmIF) on matched frozen and FFPE samples. The functional impact of C3a/C5a signaling was assessed using C3aR/C5aR antagonists (C3aR-AT/C5aR-AT) in liver cancer cells, followed by xenograft in zebrafish models to evaluate the mechanistic effects on tumor progression.Results Complement mRNA expression was significantly reduced in HCC tissue compared to adjacent non-tumor tissue in both FFPE (n=24) and frozen (n=29) samples. In contrast, proteomic analysis (n=13) and immunoblotting (n=42) revealed increased complement protein levels in tumors, except for MBL2, which was higher in non-tumor tissues. These findings were corroborated by qmIF (n=6). The MBL2 lectin array revealed elevated mannose glycans in non-tumor samples, with reduced signals observed in tumor tissues. Functional studies using C3aR-AT/C5aR-AT in liver cancer cell lines showed reduced proliferation, migration, and invasion, alongside downregulation of EMT markers (N-cadherin, vimentin) and signaling proteins (Akt, MAPK, β-catenin) ( figure 1). In vivo, zebrafish xenografts of SNU-449 human liver cancer cells treated with the C3aR antagonist (C3aR-AT) exhibited a significant reduction in tumor burden by day 4, with a ~7.3-fold decrease in tumor mass compared to vehicle-treated controls. We also observed significantly increased apoptosis and cell proliferation, suggesting a functional role of C3aR signaling in sustaining tumor growth in this model. Additionally, C3aR-AT treatment led to a ~23.9-fold increase in neutrophil infiltration within the TME, indicating enhanced myeloid cell recruitment (figure 2).Conclusions Integrated mRNA, proteomic, and functional analyses reveal complement dysregulation in HCC, with downregulated gene expression but elevated protein levels. C3a/C5a receptor signaling emerges as a key driver of progression. Pharmacological blockade of these receptors suppresses tumor growth and EMT signaling, offering a promising therapeutic approach.Abstract 49 Figure 1Pharmacologic inhibition of C3aR/C5aR impairs liver cancer cell viabilityAbstract 49 Figure 2Therapeutic role of C3aR antagonist in zebrafish xenograft model of HCC",
  "authors": [
    {
      "affiliations": [
        "Montefiore Einstein Comprehensive Cancer Center, Bronx, NY, USA"
      ],
      "name": "Divya B Kenchappa"
    },
    {
      "affiliations": [
        "Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Ajay K Singh"
    },
    {
      "affiliations": [
        "Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Yadriel Bracero"
    },
    {
      "affiliations": [
        "Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Juliana MM Gomes"
    },
    {
      "affiliations": [
        "Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Lydia Bioh"
    },
    {
      "affiliations": [
        "Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Gerardo Espinoza"
    },
    {
      "affiliations": [
        "Montefiore Medical Center/Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Xing Li"
    },
    {
      "affiliations": [
        "Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Joaquin C Sandoval"
    },
    {
      "affiliations": [
        "Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Tianyun Jiang"
    },
    {
      "affiliations": [
        "Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Lawrence Leung"
    },
    {
      "affiliations": [
        "Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Praveen Agarwal"
    },
    {
      "affiliations": [
        "Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Sofia D Oliveira"
    },
    {
      "affiliations": [
        "Albert Einstein College of Medicine, Bronx, NY, USA"
      ],
      "name": "Yvonne M Saenger"
    }
  ],
  "title": "49 Complement system alterations in hepatocellular carcinoma (HCC): insights from mRNA-proteome integration and receptor blockade studies",
  "uid": "6fd4ff45-34b3-50a1-a4e0-6b3ba38bae75"
}