Novel extragenic genomic safe harbors for precise therapeutic T-cell engineering Journal Article
| Authors: | Odak, A.; Yuan, H.; Feucht, J.; Cantu, V. A.; Mansilla-Soto, J.; Kogel, F.; Eyquem, J.; Everett, J.; Bushman, F. D.; Leslie, C. S.; Sadelain, M. |
| Article Title: | Novel extragenic genomic safe harbors for precise therapeutic T-cell engineering |
| Abstract: | Cell therapies that rely on engineered immune cells can be enhanced by achieving uniform and controlled transgene expression in order to maximize T-cell function and achieve predictable patient responses. Although they are effective, current genetic engineering strategies that use γ-retroviral, lentiviral, and transposon-based vectors to integrate transgenes, unavoidably produce variegated transgene expression in addition to posing a risk of insertional mutagenesis. In the setting of chimeric antigen receptor (CAR) therapy, inconsistent and random CAR expression may result in tonic signaling, T-cell exhaustion, and variable T-cell persistence. Here, we report and validate an algorithm for the identification of extragenic genomic safe harbors (GSH) that can be efficiently targeted for DNA integration and can support sustained and predictable CAR expression in human peripheral blood T cells. The algorithm is based on 7 criteria established to minimize genotoxicity by directing transgene integration away from functionally important genomic elements, maximize efficient CRISPR/Cas9–mediated targeting, and avert transgene silencing over time. T cells engineered to express a CD19 CAR at GSH6, which meets all 7 criteria, are curative at low cell dose in a mouse model of acute lymphoblastic leukemia, matching the potency of CAR T cells engineered at the TRAC locus and effectively resisting tumor rechallenge 100 days after their infusion. The identification of functional extragenic GSHs thus expands the human genome available for therapeutic precision engineering. © 2023 The American Society of Hematology |
| Keywords: | controlled study; human cell; nonhuman; antigen expression; animal cell; mouse; cancer immunotherapy; animal experiment; animal model; in vivo study; antineoplastic activity; in vitro study; acute lymphoblastic leukemia; algorithm; transgene; genomics; chimeric antigen receptor; gene silencing; genotoxicity; dna integration; cell engineering; human; male; article; crispr-cas9 system; adeno associated virus 6; genomic safe harbor |
| Journal Title: | Blood |
| Volume: | 141 |
| Issue: | 22 |
| ISSN: | 0006-4971 |
| Publisher: | American Society of Hematology |
| Date Published: | 2023-06-01 |
| Start Page: | 2698 |
| End Page: | 2712 |
| Language: | English |
| DOI: | 10.1182/blood.2022018924 |
| PUBMED: | 36745870 |
| PROVIDER: | scopus |
| PMCID: | PMC10273162 |
| DOI/URL: | |
| Notes: | The MSK Cancer Center Support Grant (P30 CA008748) is acknowledged in the PDF -- Corresponding author is MSK author: Michel Sadelain -- Source: Scopus |
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