{
  "abstract": "Background Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal cancers due to its aggressive biology and resistance to existing therapies. Oncofetal chondroitin sulfate (ofCS) is a tumour-restricted glycosaminoglycan broadly expressed across solid cancers but largely absent from normal adult tissues. We developed C9-based chimeric antigen receptor (CAR)-T cells targeting ofCS to overcome poor antigen specificity and the immunosuppressive tumour microenvironment (TME).Objective To optimise ofCS-targeted C9 CAR-T cell therapy for PDAC through integrated CAR design optimisation, metabolic enhancement and TME reprogramming.Design C9 and charge-optimised C9-66 CAR-T cells were engineered using a humanised ofCS-binding single-chain antibody fragment. Antitumour efficacy, functional durability and metabolic fitness were assessed in murine and patient-derived PDAC models. Enhancement strategies included inosine-mediated metabolic reprogramming, Nr5a2 overexpression and sequential TME remodelling using GLP-1R modulation, CSF-1R blockade and programmed cell death protein-1 inhibition.Results C9 CAR-T cells exhibited potent cytotoxicity, delayed tumour progression and extended survival in PDAC models. Compared with the parental construct, charge-optimised C9-66 CAR-T cells showed reduced exhaustion and more sustained activity in vivo. Inosine enhanced cytokine production, promoted central-memory differentiation and mitigated exhaustion, whereas Nr5a2 overexpression increased mitochondrial respiration and cytotoxicity. Sequential GLP-1R on–off modulation with macrophage and checkpoint blockade enhanced intratumoural CAR-T cell activity and prolonged survival. Human C9-66 CAR-T cells retained specific ofCS recognition and lysed patient-derived PDAC cells in vitro and in vivo.Conclusion C9-66 CAR-T cells with metabolic optimisation and TME reprogramming represent a tumour-specific and clinically translatable immunotherapeutic strategy for PDAC and other ofCS-expressing solid tumours.",
  "authors": [
    {
      "affiliations": [
        "Center for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai, China",
        "State Key Laboratory of Systems Medicine for Cancer, Shanghai Jiao Tong University School of Medicine, Shanghai, China"
      ],
      "name": "Ke Jiang"
    },
    {
      "affiliations": [
        "Precision Immunotherapy, Graduate Institute of Biomedical Sciences, China Medical University, Taichung, Taiwan"
      ],
      "name": "Chu-Hu Lai"
    },
    {
      "affiliations": [
        "Center for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai, China",
        "State Key Laboratory of Systems Medicine for Cancer, Shanghai Jiao Tong University School of Medicine, Shanghai, China"
      ],
      "name": "Shuang Dong"
    },
    {
      "affiliations": [
        "Precision Immunotherapy, Graduate Institute of Biomedical Sciences, China Medical University, Taichung, Taiwan",
        "Department of Medical Laboratory and Regenerative Medicine, MacKay Medical University, New Taipei, Taiwan"
      ],
      "name": "Chi-Pin Lee"
    },
    {
      "affiliations": [
        "Pancreatic Cancer Heterogeneity, INOC – Istituto Nazionale Oncologico Candiolo, IRCCS, Candiolo, Turin, Italy"
      ],
      "name": "Miriam Roberto"
    },
    {
      "affiliations": [
        "Pancreatic Cancer Heterogeneity, INOC – Istituto Nazionale Oncologico Candiolo, IRCCS, Candiolo, Turin, Italy"
      ],
      "name": "Mengli Yue"
    },
    {
      "affiliations": [
        "Pancreatic Cancer Heterogeneity, INOC – Istituto Nazionale Oncologico Candiolo, IRCCS, Candiolo, Turin, Italy"
      ],
      "name": "Serena Tondi"
    },
    {
      "affiliations": [
        "Precision Immunotherapy, Graduate Institute of Biomedical Sciences, China Medical University, Taichung, Taiwan"
      ],
      "name": "Ya-Ping Chen"
    },
    {
      "affiliations": [
        "Center for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai, China",
        "State Key Laboratory of Systems Medicine for Cancer, Shanghai Jiao Tong University School of Medicine, Shanghai, China"
      ],
      "name": "Yilin Wang"
    },
    {
      "affiliations": [
        "Pancreatic Cancer Heterogeneity, INOC – Istituto Nazionale Oncologico Candiolo, IRCCS, Candiolo, Turin, Italy"
      ],
      "name": "Preeta Ananthanarayanan"
    },
    {
      "affiliations": [
        "Center for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai, China",
        "State Key Laboratory of Systems Medicine for Cancer, Shanghai Jiao Tong University School of Medicine, Shanghai, China"
      ],
      "name": "Mei Wang"
    },
    {
      "affiliations": [
        "Department of Medicine, School of Medicine, China Medical University, Taichung, Taiwan",
        "Department of Obstetrics and Gynecology, China Medical University Hospital, Taichung, Taiwan"
      ],
      "name": "Cherry Yin-Yi Chang"
    },
    {
      "affiliations": [
        "Centre for Translational Medicine and Parasitology (CMP) at Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark",
        "VarCT Diagnostics, Frederiksberg, Denmark"
      ],
      "name": "Mette Ø Agerbaek"
    },
    {
      "affiliations": [
        "Centre for Translational Medicine and Parasitology (CMP) at Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark",
        "VAR2 Pharmaceuticals, Frederiksberg, Denmark"
      ],
      "name": "Elena E Vidal-Calvo"
    },
    {
      "affiliations": [
        "Centre for Translational Medicine and Parasitology (CMP) at Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark",
        "VAR2 Pharmaceuticals, Frederiksberg, Denmark"
      ],
      "name": "Swati Choudhary"
    },
    {
      "affiliations": [
        "Centre for Translational Medicine and Parasitology (CMP) at Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark",
        "Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, California, USA",
        "John A. Burns School of Medicine, University of Hawaii, Honolulu, Hawaii, USA"
      ],
      "name": "Thomas M Clausen"
    },
    {
      "affiliations": [
        "Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, California, USA"
      ],
      "name": "Jeffrey Esko"
    },
    {
      "affiliations": [
        "Centre for Translational Medicine and Parasitology (CMP) at Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark",
        "VAR2 Pharmaceuticals, Frederiksberg, Denmark"
      ],
      "name": "Tobias Gustavsson"
    },
    {
      "affiliations": [
        "Centre for Translational Medicine and Parasitology (CMP) at Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark"
      ],
      "name": "Thor G Theander"
    },
    {
      "affiliations": [
        "Centre for Translational Medicine and Parasitology (CMP) at Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark"
      ],
      "name": "Ali Salanti"
    },
    {
      "affiliations": [
        "Precision Immunotherapy, Graduate Institute of Biomedical Sciences, China Medical University, Taichung, Taiwan"
      ],
      "name": "Alexandra Aicher"
    },
    {
      "affiliations": [
        "Pancreatic Cancer Heterogeneity, INOC – Istituto Nazionale Oncologico Candiolo, IRCCS, Candiolo, Turin, Italy"
      ],
      "name": "Christopher Heeschen"
    }
  ],
  "title": "Multimodal C9-66 CAR-T cell immunotherapy improves outcome in preclinical models of pancreatic cancer",
  "uid": "02a98c32-fe76-530e-a2a7-96b7ff7852a0"
}