Targeting overexpressed antigens in glioblastoma via CAR T cells with computationally designed high-affinity protein...

Area of Science:

• Immunology
• Oncology
• Biotechnology

Background:

• Chimeric antigen receptor (CAR) T cell therapy shows limited efficacy in glioblastoma (GBM).
• Existing CAR constructs often use antibody-derived single-chain variable fragments (scFvs) as antigen-binding domains.
• Glioblastoma overexpresses targets like epidermal growth factor receptor (EGFR) and CD276.

Purpose of the Study:

• To develop novel CAR T cells with de novo designed protein binders for GBM treatment.
• To evaluate the therapeutic performance of these engineered CAR T cells compared to scFv-based CAR T cells.

Main Methods:

• Computationally designed de novo protein binders with high affinity for EGFR and CD276.
• Generation of CAR T cells utilizing these novel binders.
• In vitro and in vivo assessment of CAR T cell proliferation, cytokine secretion, exhaustion resistance, and anti-tumor activity.
• Bulk and single-cell transcriptional profiling to analyze CAR expression and stability.

Main Results:

• De novo designed binders enhanced CAR T cell proliferation, cytotoxic cytokine secretion, and resistance to cell exhaustion.
• Engineered CAR T cells demonstrated improved anti-tumor efficacy in vitro and in vivo.
• CARs with designed binders showed higher surface expression and increased resistance to degradation.

Conclusions:

• De novo design of protein binders offers a promising strategy to enhance CAR T cell therapy for glioblastoma.
• This approach may improve the efficacy of CAR T cell therapies for other solid tumors.
• Engineered binders represent a potential advancement over traditional scFv domains in CAR T cell design.