{
  "abstract": "Background Glioblastoma (GBM) remains uniformly lethal due to pronounced intratumoral heterogeneity and a highly immunosuppressive microenvironment that limits the efficacy of targeted therapies.Methods We engineered chimeric antigen receptor (CAR) T cells targeting Epidermal Growth Factor Receptor variant III (EGFRvIII) and armored them with a single-chain interleukin-12 (scIL12) payload. These cells were tested in syngeneic, orthotopic GBM mouse models exhibiting heterogeneous EGFRvIII expression. CAR T cells were delivered intracranially without lymphodepletion.Results Intracranial administration of scIL12-secreting CAR-T cells eradicated tumors without requiring lymphodepletion, achieving 50% long-term survival. Survival benefits depended entirely on endogenous CD8 + T cells, as efficacy was abolished in CD8-deficient hosts and unaffected by natural killer cell depletion. Notably, therapeutic efficacy was abrogated by lymphodepletion, underscoring the necessity of an intact endogenous immune response. Mechanistically, scIL12 enhanced CAR-T cell persistence and reprogrammed tumor-associated microglia, indicating potential antigen spreading through polyclonal endogenous CD8+T cell responses, which facilitate the elimination of EGFRvIII-negative tumor cells.Conclusions This study demonstrates the pleiotropic benefits of IL-12 armored CAR-T cells with improved targeting of antigen-positive tumor cells and simultaneous remodeling of the microenvironment to engage adaptive immunity against antigen-negative clones. This strategy offers a potential clinically actionable approach to improve outcomes in GBM by circumventing the need for toxic lymphodepletion and addressing tumor heterogeneity.",
  "authors": [
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Steven Shen"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "Center for Advanced Genomic Technologies, Duke University, Durham, North Carolina, USA",
        "Duke MD-PhD (MSTP) Program, Duke University, Durham, North Carolina, USA",
        "Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA"
      ],
      "name": "Aditya A Mohan"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Kelly M Hotchkiss"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Sarah Cook"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Kisha Patel"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Eliese Moelker"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Bhairavy Puviindran"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Ariel T Gonzalez"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Saif Zaidi"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Samantha Spellicy"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Adam Schwartz"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Carter Suryadevara"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Daniel Wilkinson"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Katayoun Ayasoufi"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Peter E Fecci"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "Luis Sanchez-Perez"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA"
      ],
      "name": "John Sampson"
    },
    {
      "affiliations": [
        "Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "The Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Pathology, Duke University Medical Center, Durham, North Carolina, USA",
        "Department of Neurosurgery, Duke University Medical Center, Durham, North Carolina, USA",
        "Center for Advanced Genomic Technologies, Duke University, Durham, North Carolina, USA",
        "Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA"
      ],
      "name": "Anoop Patel"
    }
  ],
  "title": "IL-12-secreting CAR-T cells reprogram the tumor microenvironment and improve efficacy against heterogeneous models of glioblastoma",
  "uid": "c7d63e19-c911-578a-a7d9-30b84d363edc"
}