SEED-Selection enables high-efficiency enrichment of primary T cells edited at multiple loci

Engineering T cell specificity and function at multiple loci can generate more effective cellular therapies, but current manufacturing methods produce heterogenous mixtures of partially engineered cells. Here we develop a one-step process to enrich unlabeled cells containing knock-ins at multiple target loci using a family of repair templates named synthetic exon expression disruptors (SEEDs). SEEDs associate transgene integration with the disruption of a paired target endogenous surface protein while preserving target expression in nonmodified and partially edited cells to enable their removal (SEED-Selection). We design SEEDs to modify three critical loci encoding T cell specificity, coreceptor expression and major histocompatibility complex expression. The results demonstrate up to 98% purity after selection for individual modifications and up to 90% purity for six simultaneous edits (three knock-ins and three knockouts). This method is compatible with existing clinical manufacturing workflows and can be readily adapted to other loci to facilitate production of complex gene-edited cell therapies.

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Publication Details

Journal
Nature Biotechnology
Published
2025-02-05
DOI
https://doi.org/10.1038/s41587-024-02531-6
Citations
12
Primary Topic
CAR-T cell therapy research
Type
article
Field-Weighted Citation Impact
12.54

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article

SEED-Selection enables high-efficiency enrichment of primary T cells edited at multiple loci

Chun Jimmie Ye, Chang Liu, Joseph J. Muldoon, Charlotte H. Wang et al.
12 citations
Nature Biotechnology
CAR-T cell therapy research
12.54
article

SEED-Selection enables high-efficiency enrichment of primary T cells edited at multiple loci

Chun Jimmie Ye, Chang Liu, Joseph J. Muldoon, Charlotte H. Wang, Vivasvan S. Vykunta, Thomas Mazumder, Justin Eyquem, Jae Hyun J. Lee, Brian R. Shy, Vincent Allain, Daniel B. Goodman, Alexander Marson, William A. Nyberg, Allison Rothrock, Clara Kochendoerfer, Lian Li, Yang Sun, Christopher Chang
article en
12 citations

Abstract

Engineering T cell specificity and function at multiple loci can generate more effective cellular therapies, but current manufacturing methods produce heterogenous mixtures of partially engineered cells. Here we develop a one-step process to enrich unlabeled cells containing knock-ins at multiple target loci using a family of repair templates named synthetic exon expression disruptors (SEEDs). SEEDs associate transgene integration with the disruption of a paired target endogenous surface protein while preserving target expression in nonmodified and partially edited cells to enable their removal (SEED-Selection). We design SEEDs to modify three critical loci encoding T cell specificity, coreceptor expression and major histocompatibility complex expression. The results demonstrate up to 98% purity after selection for individual modifications and up to 90% purity for six simultaneous edits (three knock-ins and three knockouts). This method is compatible with existing clinical manufacturing workflows and can be readily adapted to other loci to facilitate production of complex gene-edited cell therapies.

Nature BiotechnologyVol. 43(12)
Gladstone Institutes (US), Inserm (FR), University of California, San Francisco (US), Université Paris Cité (FR), Innovative Genomics Institute (US), Parker Institute for Cancer Immunotherapy (US), Palo Alto Institute (US), Hôpital Saint-Louis (FR), UCSF Helen Diller Family Comprehensive Cancer Center (US), Arc Research Institute, University of California, Berkeley (US)
Cancer Research Institute, National Institute of General Medical Sciences
Openalex Percentile: Top 1%
CAR-T cell therapy research
12.54
DOIOpen Access PDFOpenAlex
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