Mesothelin-based CAR-T cells exhibit potent antitumor activity against ovarian cancer

• Published in: Journal of translational medicine 2024-04, Vol.22 (1), p.367-367, Article 367
• Main Authors: Guo, Jing, Zeng, Xiaozhu, Zhu, Yongjie, Yang, Dong, Zhao, Xudong
• Format: Article
• Language: English
• Subjects: Analysis; Animals; Apoptosis; CAR-T cells; Care and treatment; Cell Line, Tumor; Diagnosis; Female; Fluorescent antibody technique; GPI-Linked Proteins - metabolism; Humans; Immunofluorescence; Mesothelin; Mice; MUC16 (CA125); Mucins; Ovarian cancer; Ovarian Neoplasms - drug therapy; T-Lymphocytes - metabolism; Testing; Xenograft Model Antitumor Assays
• ISSN: 1479-5876; 1479-5876
• DOI: 10.1186/s12967-024-05174-y

Ovarian cancer (OC) is characterized by its rapid growth and spread which, accompanied by a low 5-year survival rate, necessitates the development of improved treatments. In ovarian cancer, the selective overexpression of Mucin-16 (MUC16, CA125) in tumor cells highlights its potential as a promising target for developing anti-tumor therapies. However, the potential effectiveness of CAR-T cell therapy that targets MUC16 in ovarian cancer cells is unknown. The expression of MUC16 in viable OC cells was detected using immunofluorescence and flow cytometry techniques. A MSLN-CAR construct, comprising the MUC16-binding polypeptide region of mesothelin (MSLN), a CD8 hinge spacer and transmembrane domain, 4-1BB, and CD3ζ endo-domains; was synthesized and introduced into T cells using lentiviral particles. The cytotoxicity of the resultant CAR-T cells was evaluated in vitro using luciferase assays. Cytokine release by CAR-T cells was measured using enzyme-linked immunosorbent assays. The anti-tumor efficacy of the CAR-T cells was subsequently assessed in mice through both systemic and local administration protocols. MSLN-CAR T cells exhibited potent cytotoxicity towards OVCAR3 cells and their stem-like cells that express high levels of MUC16. Also, MSLN-CAR T cells were inefficient at killing SKOV3 cells that express low levels of MUC16, but were potently cytotoxic to such cells overexpressing MUC16. Moreover, MSLN-CAR T cells delivered via tail vein or peritoneal injection could shrink OVCAR3 xenograft tumors in vivo, with sustained remission observed following peritoneal delivery of MSLN-CAR T cells. Collectively, these results suggested that MSLN-CAR T cells could potently eliminate MUC16- positive ovarian cancer tumor cells both in vitro and in vivo, thereby providing a promising therapeutic intervention for MUC16-positive patients.