CAR-T 療法：癌症治療的未來

CAR-T cell therapy involves a multi-step manufacturing process and specific molecular architecture:

• Autologous Manufacturing Process:
  • T-cell Isolation: Patient's T cells are collected from blood via leukapheresis.
  • T-cell Activation: Isolated T cells are activated ex vivo.
  • Genetic Modification: T cells are genetically engineered to express Chimeric Antigen Receptors (CARs). This typically involves introducing a CAR transgene using viral vectors (e.g., lentivirus) or non-viral methods such as CRISPR/Cas9, transposons, or mRNA transfection.
  • Ex Vivo Expansion: The modified CAR-T cells are expanded in a controlled environment to achieve sufficient cell numbers for therapeutic dosing.
  • Formulation and Cryopreservation: The expanded CAR-T cells are formulated and often cryopreserved before being infused back into the patient.
• CAR Structure: A CAR is a synthetic receptor protein composed of four main components:
  • Extracellular Target Antigen Binding Domain: Typically a single-chain variable fragment (scFv) derived from monoclonal antibodies, responsible for recognizing and binding to specific antigens on cancer cells.
  • Hinge/Spacer Region: Provides flexibility and optimal distance for antigen binding.
  • Transmembrane Domain: Anchors the CAR to the T cell membrane.
  • Intracellular Signaling Domains: One or more domains (e.g., CD3ζ, CD28, 4-1BB/CD137) that transmit activation signals inside the T cell upon antigen binding, promoting T-cell proliferation and effector functions.
• CAR Generations:
  • First-generation: Contained only a CD3ζ or FcRγ signaling domain, with limited persistence.
  • Second-generation: Include one co-stimulatory domain (e.g., CD28 or 4-1BB) in addition to CD3ζ, significantly enhancing T-cell persistence and anti-tumor activity.
  • Third-generation: Incorporate two co-stimulatory domains, though preclinical results have been mixed.
  • Fourth-generation (e.g., 'Armored CARs'): Designed to secrete additional cytokines (e.g., IL-12 or IL-18) to further enhance T-cell activation, recruit other immune cells, and overcome the immunosuppressive tumor microenvironment, particularly relevant for solid tumors.
• Key Target Antigens: Approved CAR-T therapies primarily target CD19 for B-cell malignancies and BCMA for multiple myeloma. Emerging targets for solid tumors and other leukemias include GPC3 (liver cancer), GPC2 (neuroblastoma), CD7 (T-cell malignancies), HER2, EGFR, NKG2D, and mesothelin.