Non-viral therapeutic cell engineering with the Sleeping Beauty transposon system

Hudecek, Michael; Ivics, Zoltán

doi:10.1016/j.gde.2018.06.003

Cited by 48 publications

(37 citation statements)

References 85 publications

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“…The advantages of using the SB system for gene therapy include the ease and reduced cost of manufacturing of clinical-grade, plasmid-based vectors compared to recombinant viral vectors [36], scalability of vector production, the ease of ensuring quality control for clinical use, and indefinite storage with absolute fidelity. In addition, there is pronounced flexibility with respect to a wide range of human cell types that are amenable to SB transposon-based nonviral gene delivery and with respect to a wide variety of transgene cassettes that can be efficiently incorporated into the target cell genome by this system.…”

Section: Discussionmentioning

confidence: 99%

Genomic Analyses of SLAMF7 CAR-T Cells Manufactured by Sleeping Beauty Transposon Gene Transfer for Immunotherapy of Multiple Myeloma

Miskey

Amberger

Reiser

et al. 2019

Preprint

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Widespread treatment of human diseases with gene therapies necessitates the development of gene transfer vectors that integrate genetic information effectively, safely and economically. Accordingly, significant efforts have been devoted to engineer novel tools that i) achieve high-level stable gene transfer at low toxicity to the host cell; ii) induce low levels of genotoxicity and possess a ‘safe’ integration profile with a high proportion of integrations into safe genomic locations; and iii) are associated with acceptable cost per treatment and scalable/exportable vector production to serve large numbers of patients. The Sleeping Beauty (SB) transposon has been transformed into a vector system that is fulfilling these requirements.In the CARAMBA project, we use SB transposition to genetically modify T cells with a chimeric antigen receptor (CAR) specific for the SLAMF7 antigen, that is uniformly and highly expressed on malignant plasma cells in multiple myeloma. We have demonstrated that SLAMF7 CAR-T cells confer specific and very potent anti-myeloma reactivity in pre-clinical models, and are therefore preparing a Phase I/IIa clinical trial of adoptive immunotherapy with autologous, patient-derived SLAMF7-CAR T cells in multiple myeloma (EudraCT Nr. 2019-001264-30/CARAMBA-1).Here we report on the characterization of genomic safety attributes in SLAMF7 CAR-T cells that we prepared in three clinical-grade manufacturing campaigns under good manufacturing practice (GMP), using T cells that we obtained from three healthy donor volunteers. In the SLAMF7 CAR-T cell product, we determined the average transposon copy number, the genomic insertion profile, and presence of residual SB100X transposase. The data show that the SLAMF7 CAR transposon had been inserted into the T cell genome with the close-to-random distribution pattern that is typical for SB, and with an average transposon copy number ranging between 6 and 12 per T cell. No residual SB100X transposase could be detected by Western blotting in the infusion products. With these attributes, the SLAMF7 CAR-T products satisfy criteria set forth by competent regulatory authorities in order to justify administration of SLAMF7 CAR-T cells to humans in the context of a clinical trial. These data set the stage for the CARAMBA clinical trial, that will be the first in the European Union to use virus-free SB transposition for CAR-T engineering.DisclosuresThis project is receiving funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 754658 (CARAMBA).

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Section: Discussionmentioning

confidence: 99%

Genomic Analyses of SLAMF7 CAR-T Cells Manufactured by Sleeping Beauty Transposon Gene Transfer for Immunotherapy of Multiple Myeloma

Miskey

Amberger

Reiser

et al. 2019

Preprint

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“…n = 1, or no indication at all (unknown) n = 80; Figure 4). Some clinical trials use a non-viral gene delivery system or a transfer method integrating the CAR gene into a specified site (i.e., sleeping beauty transposon system [103,[105][106][107], PiggyBac transposon system [103,107], CRISPR-Cas9 [108], or transfection of DNA or RNA [109]). The latter two methods do not result in an integration of the CAR-encoding gene into the host cell's genome, which has certain advantages (highlighted in Section 4.3 for mRNA transfection).…”

Section: Transfer Methods To Introduce the Car Into T Cellsmentioning

confidence: 99%

The Landscape of CAR-T Cell Clinical Trials against Solid Tumors—A Comprehensive Overview

Schaft

2020

Cancers

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CAR-T cells showed great potential in the treatment of patients with hematologic tumors. However, the clinical efficacy of CAR-T cells against solid tumors lags behind. To obtain a comprehensive overview of the landscape of CAR-T cell clinical trials against this type of cancer, this review summarizes all the 196 studies registered at clinicaltrials.gov. Special focus is on: (1) geographical distribution; (2) targeted organs, tumor entities, and antigens; (3) CAR transfer methods, CAR formats, and extra features introduced into the T cells; and (4) patient pretreatments, injection sites, and safety measurements. Finally, the few data on clinical outcome are reported. The last assessment of clinicaltrials.gov for the data summarized in this paper was on 4 August 2020.

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“…Sleeping Beauty (SB) transposon vectors have provided a safe, effective and economically method to integrate genetic information through gene transfer vectors and they may overcome shortcomings of viral vectors [47,48]. SB transposon system has been successfully applied to CAR-T cell therapy.…”

Section: Car Transduction Into Nk Cellsmentioning

confidence: 99%

Chimeric antigen receptor (CAR)-modified NK cells against cancer: Opportunities and challenges

Wang

Dou

et al. 2019

International Immunopharmacology

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Non-viral therapeutic cell engineering with the Sleeping Beauty transposon system

Cited by 48 publications

References 85 publications

Genomic Analyses of SLAMF7 CAR-T Cells Manufactured by Sleeping Beauty Transposon Gene Transfer for Immunotherapy of Multiple Myeloma

Genomic Analyses of SLAMF7 CAR-T Cells Manufactured by Sleeping Beauty Transposon Gene Transfer for Immunotherapy of Multiple Myeloma

The Landscape of CAR-T Cell Clinical Trials against Solid Tumors—A Comprehensive Overview

Chimeric antigen receptor (CAR)-modified NK cells against cancer: Opportunities and challenges

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