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Related Concept Videos

Gene Therapy00:59

Gene Therapy

Gene therapy is a technique where a gene is inserted into a person’s cells to prevent or treat a serious disease. The added gene may be a healthy version of the gene that is mutated in the patient, or it could be a different gene that inactivates or compensates for the patient’s disease-causing gene. For example, in patients with severe combined immunodeficiency (SCID) due to a mutation in the gene for the enzyme adenosine deaminase, a functioning version of the gene can be inserted. The...

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An Engineered Adeno-Associated Virus Variant Enables Efficient Gene Editing in Human T Cells.

Mi Leng1, Chunmei Gan1, Zhaoyue Zheng2

  • 1National Clinical Research Center for Geriatrics and State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.

Human Gene Therapy
|February 26, 2026
PubMed
Summary

Researchers engineered a novel recombinant adeno-associated virus (rAAV) variant, Tot3, to improve chimeric antigen receptor T (CAR-T) cell therapy for leukemia. Tot3 enhances gene editing efficiency in T cells, leading to better antitumor activity in preclinical models.

Keywords:
B cell acute lymphocytic leukemiacapsid proteinchimeric antigen receptor T cell immunotherapygene editingrecombinant adeno-associated virus

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Area of Science:

  • Molecular Biology
  • Immunotherapy
  • Gene Therapy

Background:

  • Recombinant adeno-associated viruses (rAAVs) are crucial for gene editing in chimeric antigen receptor T (CAR-T) cell therapy.
  • Current rAAV serotypes have limitations in efficiently transducing primary T cells at low multiplicities of infection (MOIs).
  • Developing improved rAAV vectors is essential for advancing CAR-T cell therapy for leukemia and other cancers.

Purpose of the Study:

  • To identify and characterize novel rAAV variants with enhanced transduction efficiency in human primary T cells.
  • To evaluate the efficacy of a new rAAV variant, Tot3, for gene editing in CAR-T cells.
  • To assess the therapeutic potential of Tot3-modified CAR-T cells in a preclinical model of diffuse B cell lymphoma.

Main Methods:

  • Screening of an AAV2 peptide library in Jurkat cells to identify high-performing variants.
  • Validation of selected variants, including Tot3, in primary human T cells.
  • Gene editing of programmed cell death protein 1 (PD-1) and CAR overexpression in T cells using Tot3.
  • Assessment of CAR-T cell antitumor activity and survival in a mouse model.

Main Results:

  • A novel rAAV variant, Tot3, was identified with transduction efficiency comparable to AAV2 but at a 27-fold lower MOI.
  • Tot3 demonstrated increased packaging efficiency and reduced thermal stability compared to AAV2.
  • High knockout and knock-in efficiencies (up to 70% and 55%) were achieved for PD-1 and CAR, respectively, in primary T cells.
  • Tot3-modified CAR-T cells exhibited significantly enhanced antitumor activity and prolonged survival in a diffuse B cell lymphoma mouse model.

Conclusions:

  • Tot3 represents a promising rAAV variant for efficient gene editing in human primary T cells.
  • This advancement can improve the manufacturing and efficacy of CAR-T cell therapies.
  • Tot3 holds potential for enhancing the treatment of hematological malignancies like leukemia and lymphoma.