Allogeneic CRISPR-Engineered CAR-T Cells Drive Potent Antitumor Activity in Solid Tumors

Area of Science:

• Immunology
• Oncology
• Gene Therapy

Background:

• Chimeric antigen receptor (CAR) T-cell therapy shows limited efficacy in solid tumors due to patient-specific T-cell variability.
• Advanced disease and prior treatments in patients can compromise autologous T-cell quality for CAR-T therapy.

Purpose of the Study:

• To develop an "off-the-shelf" allogeneic CAR-T cell platform for solid tumors.
• To engineer CAR-T cells targeting glypican-2 (GPC2) and glypican-3 (GPC3) using CRISPR-Cas9 genome editing.

Main Methods:

• Utilized CRISPR-Cas9 to insert CAR into the TRAC locus and disrupt B2M in healthy donor T cells.
• Employed adeno-associated virus (AAV) for CAR delivery targeting GPC2 and GPC3.
• Evaluated antigen-specific cytotoxicity and in vivo efficacy in preclinical solid tumor models.

Main Results:

• Genome-edited allogeneic CAR-T cells demonstrated potent, antigen-specific cytotoxicity against multiple tumor models.
• GPC2-directed CAR-T cells showed enhanced activity in neuroblastoma models, leading to tumor regression and prolonged survival.
• GPC3-directed CAR-T cells exhibited robust activity against hepatocellular carcinoma in vitro and in vivo.
• Repeated dosing of CAR-T cells increased efficacy without observed toxicity.

Conclusions:

• Established a scalable, genome-engineered allogeneic CAR-T strategy for solid tumors.
• The platform shows strong therapeutic potential, supporting clinical development for pediatric and adult solid tumors.
• Off-the-shelf allogeneic CAR-T cells offer a promising alternative to autologous therapies.