Modular pooled discovery of synthetic knockin sequences to program durable cell therapies DOI Creative Commons
Franziska Blaeschke, Yan Yi Chen, Ryan Apathy

et al.

Cell, Journal Year: 2023, Volume and Issue: 186(19), P. 4216 - 4234.e33

Published: Sept. 1, 2023

Chronic stimulation can cause T cell dysfunction and limit the efficacy of cellular immunotherapies. Improved methods are required to compare large numbers synthetic knockin (KI) sequences reprogram functions. Here, we developed modular pooled KI screening (ModPoKI), an adaptable platform for construction DNA libraries using barcoded multicistronic adaptors. We built two ModPoKI 100 transcription factors (TFs) 129 natural surface receptors (SRs). Over 30 screens across human TCR- CAR-T cells in diverse conditions identified a factor AP4 (TFAP4) construct that enhanced fitness chronically stimulated anti-cancer function vitro vivo. ModPoKI's modularity allowed us generate ∼10,000-member library TF combinations. Non-viral combined BATF-TFAP4 polycistronic fitness. Overexpressed BATF TFAP4 co-occupy regulate key gene targets function. facilitates discovery complex constructs program

Language: Английский

CRISPR screens for functional interrogation of immunity DOI
Hao Shi, John G. Doench, Hongbo Chi

et al.

Nature reviews. Immunology, Journal Year: 2022, Volume and Issue: 23(6), P. 363 - 380

Published: Dec. 8, 2022

Language: Английский

Citations

53
Production and characterization of virus-free, CRISPR-CAR T cells capable of inducing solid tumor regression DOI Creative Commons
Katherine P. Mueller,

Nicole J. Piscopo,

Matthew H. Forsberg

et al.

Journal for ImmunoTherapy of Cancer, Journal Year: 2022, Volume and Issue: 10(9), P. e004446 - e004446

Published: Sept. 1, 2022

Chimeric antigen receptor (CAR) T cells have demonstrated high clinical response rates against hematological malignancies (e.g., CD19+ cancers) but shown limited activity in patients with solid tumors. Recent work showed that precise insertion of a CAR at defined locus improves treatment outcomes the context CD19 CAR; however, it is unclear if such strategy could also affect Furthermore, manufacturing generally relies on viral vectors for gene delivery, which comprise complex and resource-intensive part supply chain.

Language: Английский

Citations

40
Genetic engineering meets hematopoietic stem cell biology for next-generation gene therapy DOI Creative Commons
Samuele Ferrari, Erika Valeri, Anastasia Conti

et al.

Cell stem cell, Journal Year: 2023, Volume and Issue: 30(5), P. 549 - 570

Published: May 1, 2023

Language: Английский

Citations

37
Combining different CRISPR nucleases for simultaneous knock-in and base editing prevents translocations in multiplex-edited CAR T cells DOI Creative Commons
Viktor Glaser, Christian Flugel, Jonas Kath

et al.

Genome biology, Journal Year: 2023, Volume and Issue: 24(1)

Published: April 24, 2023

Multiple genetic modifications may be required to develop potent off-the-shelf chimeric antigen receptor (CAR) T cell therapies. Conventional CRISPR-Cas nucleases install sequence-specific DNA double-strand breaks (DSBs), enabling gene knock-out or targeted transgene knock-in. However, simultaneous DSBs provoke a high rate of genomic rearrangements which impede the safety edited cells.

Language: Английский

Citations

31
Modular pooled discovery of synthetic knockin sequences to program durable cell therapies DOI Creative Commons
Franziska Blaeschke, Yan Yi Chen, Ryan Apathy

et al.

Cell, Journal Year: 2023, Volume and Issue: 186(19), P. 4216 - 4234.e33

Published: Sept. 1, 2023

Chronic stimulation can cause T cell dysfunction and limit the efficacy of cellular immunotherapies. Improved methods are required to compare large numbers synthetic knockin (KI) sequences reprogram functions. Here, we developed modular pooled KI screening (ModPoKI), an adaptable platform for construction DNA libraries using barcoded multicistronic adaptors. We built two ModPoKI 100 transcription factors (TFs) 129 natural surface receptors (SRs). Over 30 screens across human TCR- CAR-T cells in diverse conditions identified a factor AP4 (TFAP4) construct that enhanced fitness chronically stimulated anti-cancer function vitro vivo. ModPoKI's modularity allowed us generate ∼10,000-member library TF combinations. Non-viral combined BATF-TFAP4 polycistronic fitness. Overexpressed BATF TFAP4 co-occupy regulate key gene targets function. facilitates discovery complex constructs program

Language: Английский

Citations

27