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  • Highlights in Immuno-Oncology
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Serving up whatever you wish: CRISPR-base editing generates novel cancer-restricted antigens for immunotherapy

Genes & Immunity volume 24, pages 292–294 (2023)Cite this article

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Casirati et al. focused on the acute myeloid leukemia (AML) markers FLT3, CD123, and KIT, and Wellhausen et al. on the prototypic hematopoietic lineage antigen, CD45. Both groups generated CAR constructs and performed epitope mapping to identify the respective target epitopes. Then, both turned to the CRISPR base-editing technique [6] to introduce non-synonymous mutations, due to either A > G or C > T transitions with adenine base editors or cytosine base editors, respectively, that would void the CAR T cell epitopes. CRISPR base-editing waives the reliance on the DNA double strand breaks/homology-directed repair mechanism or donor DNA templates and their associated risks. With selection of the right combination of guide RNAs and base-editing enzymes, a single amino acid substitution that was sufficient to prevent recognition by the respective CAR T cells while maintaining functionality of the modified target protein could be installed in all four markers. On-target editing efficiencies were high (79% to over 90%) and non-edited cells retained the expression of the unedited target molecule, throwing them to the wolves for in vivo selection. Off-target effects and “bystander mutations” were found, but of minimal impact, and compared to the genotoxic risk of conventional chemotherapy or the risk of disease progression, Casirati et al. considered the potential harm to be minor.

The resulting stealth receptors were fully functional and epitope editing of HSCs did not affect their stemness and differentiation ability or the functionality of their lineage-specific progenies. For CD45, also the CAR T cells themselves were epitope-edited to prevent on-target/off-tumor toxicity in the form of fratricide. In silico prediction and in vitro assays suggested that the amino acid change in CD45 would not be immunogenic. Immunosuppressive graft-versus-host disease prophylaxis may additionally confer tolerance towards the epitope edited molecules.

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Fig. 1: CRISPR-base editing generates novel cancer-restricted antigens.

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Funding

Grant support: ACIR is a not-for-profit organization supported by generous philanthropic donations from individuals and industry.

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Authors and Affiliations

  1. Accelerating Cancer Immunotherapy Research, Concord, MA, USA

    Ute E. Burkhardt & Edward F. Fritsch

  2. Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA

    Edward F. Fritsch

  3. Broad Institute of MIT and Harvard, Cambridge, MA, USA

    Edward F. Fritsch

Authors
  1. Ute E. Burkhardt
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  2. Edward F. Fritsch
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Contributions

Writing, reviewing and/or revision of the manuscript: EFF, UEB.

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Correspondence to Ute E. Burkhardt or Edward F. Fritsch.

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Competing interests

EFF is an equity holder and consultant for BioNTech, and equity holder and SAB member of BioEntre, and equity holder in Dionis Therapeutics. UEB is an equity holder in BioEntre, BioNTech and Dionis Therapeutics.

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Burkhardt, U.E., Fritsch, E.F. Serving up whatever you wish: CRISPR-base editing generates novel cancer-restricted antigens for immunotherapy. Genes Immun 24, 292–294 (2023). https://doi.org/10.1038/s41435-023-00227-6

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  • DOI: https://doi.org/10.1038/s41435-023-00227-6

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