In vivo human T cell engineering with enveloped delivery vehicles

Jennifer R Hamilton; Evelyn Chen; Barbara S Perez; Cindy R Sandoval Espinoza; Min Hyung Kang; Marena Trinidad; Wayne Ngo; Jennifer A Doudna

doi:10.1038/s41587-023-02085-z

In vivo human T cell engineering with enveloped delivery vehicles

Nat Biotechnol. 2024 Jan 11. doi: 10.1038/s41587-023-02085-z. Online ahead of print.

Authors

Jennifer R Hamilton^{1

2

3}, Evelyn Chen^{1

2

3}, Barbara S Perez^{1

2}, Cindy R Sandoval Espinoza^{1

2}, Min Hyung Kang^{1

2}, Marena Trinidad^{1

2}, Wayne Ngo^{4

5}, Jennifer A Doudna^{6

7

8

9

10

11

12

13}

Affiliations

¹ Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
² Innovative Genomics Institute, University of California, Berkeley, CA, USA.
³ Azalea Therapeutics, Berkeley, CA, USA.
⁴ Gladstone Institutes, San Francisco, CA, USA.
⁵ California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA, USA.
⁶ Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA. doudna@berkeley.edu.
⁷ Innovative Genomics Institute, University of California, Berkeley, CA, USA. doudna@berkeley.edu.
⁸ Gladstone Institutes, San Francisco, CA, USA. doudna@berkeley.edu.
⁹ California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA, USA. doudna@berkeley.edu.
¹⁰ Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, USA. doudna@berkeley.edu.
¹¹ Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA. doudna@berkeley.edu.
¹² Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA. doudna@berkeley.edu.
¹³ Department of Chemistry, University of California, Berkeley, Berkeley, CA, USA. doudna@berkeley.edu.

PMID: 38212493
DOI: 10.1038/s41587-023-02085-z

Abstract

Viruses and virally derived particles have the intrinsic capacity to deliver molecules to cells, but the difficulty of readily altering cell-type selectivity has hindered their use for therapeutic delivery. Here, we show that cell surface marker recognition by antibody fragments displayed on membrane-derived particles encapsulating CRISPR-Cas9 protein and guide RNA can deliver genome editing tools to specific cells. Compared to conventional vectors like adeno-associated virus that rely on evolved capsid tropisms to deliver virally encoded cargo, these Cas9-packaging enveloped delivery vehicles (Cas9-EDVs) leverage predictable antibody-antigen interactions to transiently deliver genome editing machinery selectively to cells of interest. Antibody-targeted Cas9-EDVs preferentially confer genome editing in cognate target cells over bystander cells in mixed populations, both ex vivo and in vivo. By using multiplexed targeting molecules to direct delivery to human T cells, Cas9-EDVs enable the generation of genome-edited chimeric antigen receptor T cells in humanized mice, establishing a programmable delivery modality with the potential for widespread therapeutic utility.

Grants and funding

K99 GM143461/GM/NIGMS NIH HHS/United States