Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection

By Justin Eyquem, J. Mansilla-Soto, Theodoros Giavridis, Sjoukje J. C. van der Stegen, Mohamad Hamieh, K. Cunanan, Ashlesha Odak, M. Gönen, Michel Sadelain. Nature, 2017

1,655 citations recorded by the source corpus.

Chimeric antigen receptors (CARs) are synthetic receptors that redirect and reprogram T cells to mediate tumour rejection. The most successful CARs used to date are those targeting CD19 (ref. 2), which offer the prospect of complete remission in patients with chemorefractory or relapsed B-cell malignancies. CARs are typically transduced into the T cells of a patient using γ-retroviral vectors or other randomly integrating vectors, which may result in clonal expansion, oncogenic transformation, variegated transgene expression and transcriptional silencing. Recent advances in genome editing enable efficient sequence-specific interventions in human cells, including targeted gene delivery to the CCR5 and AAVS1 loci. Here we show that directing a CD19-specific CAR to the T-cell receptor α constant (TRAC) locus not only results in uniform CAR expression in human peripheral blood T cells, but also enhances T-cell potency, with edited cells vastly outperforming conventionally generated CAR T cells in a mouse model of acute lymphoblastic leukaemia. We further demonstrate that targeting the CAR to the TRAC locus averts tonic CAR signalling and establishes effective internalization and re-expression of the CAR following single or repeated exposure to antigen, delaying effector T-cell differentiation and exhaustion. These findings uncover facets of CAR immunobiology and underscore the potential of CRISPR/Cas9 genome editing to advance immunotherapies.

Subfields: Electrical and Electronic Engineering, Molecular Biology, Oncology

View Paper: https://www.semanticscholar.org/paper/47af46911569825ab86ac946a5c9cd502504aa3f

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Citations in the corpus

Mesothelin-targeted CAR-T cell therapy for solid tumors (2020, background): “There is also interest in viral-based targeted integration into the T cell receptor locus [98].”
Identification of Pre-Existing Adaptive Immunity to Cas9 Proteins in Humans (2018, positive): “These techniques are being applied to create novel cell therapies, either through ex vivo editing of cells (4-6) followed by the transplantation of engineered cells into a patient, or through in vivo editing of the patient’s cells by delivery of Cas9/gRNA via viral vectors (mainly with recombinant a”
CAR T Cells for Solid Tumors: New Strategies for Finding, Infiltrating, and Surviving in the Tumor Microenvironment (2019, methodology): “A study with a CD19/28ζ CAR used CRISPR to insert the CAR under the control of the TCR promoter (at the TRAC locus), while simultaneously knocking out the TCR via insertion of the CAR gene (182).”
Gene therapy comes of age (2018, positive): “In addition, the regulation of an introduced or corrected gene will be controlled by the endogenous promoter, resulting in more physiologic and appropriately regulated gene expression (63).”
Engineering strategies to overcome the current roadblocks in CAR T cell therapy (2019, methodology): “…gene construct into the TRAC locus has also been achieved using CRISPR– Cas9 in preclinical models, thereby knocking out the TCR and inserting the CAR in one step 238 ; this feat has also been achieved using an engineered I-CreI homing endonuclease and an adeno-associated virus donor template 239 .”