Bispecific ANXA2/CD147 CAR-T cell therapy for osteosarcoma

Area of Science:

• Immunology
• Oncology
• Biotechnology

Background:

• Osteosarcoma is a malignant bone cancer with poor prognosis.
• Current single-target CAR-T cell therapies for osteosarcoma have limited efficacy.
• There is a need for improved therapeutic strategies for osteosarcoma.

Purpose of the Study:

• To develop and evaluate a novel bispecific CAR-T cell therapy for osteosarcoma.
• To identify and validate suitable antigens for bispecific targeting in osteosarcoma.
• To assess the in vitro and in vivo efficacy of the bispecific CAR-T cells.

Main Methods:

• Single-cell RNA sequencing (scRNA-seq) identified ANXA2 and CD147 as target antigens.
• Bispecific ANXA2/CD147 CAR-T constructs were generated via lentiviral transduction.
• In vitro cytotoxicity assays (LDH release) and cytokine analysis (ELISA) were performed.
• In vivo efficacy was evaluated in subcutaneous CDX, xenograft, and PDX osteosarcoma models.
• Tumor immune microenvironment changes, including macrophage infiltration, were analyzed.

Main Results:

• Bispecific CAR-T cells demonstrated significantly enhanced cytotoxicity compared to single-target CAR-T cells.
• Elevated levels of effector molecules (GZMB, TNFα) were released by bispecific CAR-T cells.
• In vivo models showed superior tumor suppression, reduced metastasis, and increased T-cell infiltration.
• Increased infiltration of M1 macrophages was observed in tumors treated with bispecific CAR-T cells.

Conclusions:

• A bispecific ANXA2/CD147 CAR-T cell therapy was successfully developed.
• This therapy exhibits robust in vitro and in vivo antitumor activity against osteosarcoma.
• Bispecific CAR-T cells represent a promising new strategy for osteosarcoma immunotherapy.