Cell membrane-anchored and tumor-targeted IL-12 T-cell therapy destroys cancer-associated fibroblasts and disrupts extracellular matrix in heterogenous osteosarcoma xenograft models

• Published in: Journal for immunotherapy of cancer Vol. 12; no. 1; p. e006991
• Main Authors: Hu, Jiemiao, Lazar, Alexander J, Ingram, Davis, Wang, Wei-Lien, Zhang, Wendong, Jia, Zhiliang, Ragoonanan, Dristhi, Wang, Jian, Xia, Xueqing, Mahadeo, Kris, Gorlick, Richard, Li, Shulin
• Format: Journal Article
• Language: English
• Published: England BMJ Publishing Group Ltd 09.01.2024; BMJ Publishing Group
• Series: Original research
• Subjects: Animals; Bone Neoplasms - therapy; Cancer-Associated Fibroblasts; Cell Membrane; Cell- and Tissue-Based Therapy; Clinical/Translational Cancer Immunotherapy; Cytokines; Disease Models, Animal; Extracellular Matrix; Heterografts; Humans; Immunotherapy, Adoptive; Interleukin-12; Lymphocytes, Tumor-Infiltrating; Osteosarcoma - therapy; Pediatrics; Tumor Microenvironment; Tumor Microenvironment; Pediatrics; Lymphocytes, Tumor-Infiltrating; Cytokines; Immunotherapy, Adoptive
• ISSN: 2051-1426; 2051-1426
• DOI: 10.1136/jitc-2023-006991

BackgroundThe extracellular matrix (ECM) and cancer-associated fibroblasts (CAFs) play major roles in tumor progression, metastasis, and the poor response of many solid tumors to immunotherapy. CAF-targeted chimeric antigen receptor-T cell therapy cannot infiltrate ECM-rich tumors such as osteosarcoma.MethodIn this study, we used RNA sequencing to assess whether the recently invented membrane-anchored and tumor-targeted IL-12-armed (attIL12) T cells, which bind cell-surface vimentin (CSV) on tumor cells, could destroy CAFs to disrupt the ECM. We established an in vitro model of the interaction between osteosarcoma CAFs and attIL12-T cells to uncover the underlying mechanism by which attIL12-T cells penetrate stroma-enriched osteosarcoma tumors.ResultsRNA sequencing demonstrated that attIL12-T cell treatment altered ECM-related gene expression. Immunohistochemistry staining revealed disruption or elimination of high-density CAFs and ECM in osteosarcoma xenograft tumors following attIL12-T cell treatment, and CAF/ECM density was inversely correlated with T-cell infiltration. Other IL12-armed T cells, such as wild-type IL-12-targeted or tumor-targeted IL-12-T cells, did not disrupt the ECM because this effect depended on the engagement between CSV on the tumor cell and its ligand on the attIL12-T cells. Mechanistic studies found that attIL12-T cell treatment elevated IFNγ production on interacting with CSV+ tumor cells, suppressing transforming growth factor beta secretion and in turn upregulating FAS-mediated CAF apoptosis. CAF destruction reshaped the tumor stroma to favor T-cell infiltration and tumor inhibition.ConclusionsThis study unveiled a novel therapy—attIL12-T cells—for targeting CAFs/ECM. These findings are highly relevant to humans because CAFs are abundant in human osteosarcoma.