Safe engineering of CAR T cells for adoptive cell therapy of cancer using long‐term episomal gene transfer

Abstract Chimeric antigen receptor (CAR) T‐cell therapy is a new successful treatment for refractory B‐cell leukemia. Successful therapeutic outcome depends on long‐term expression of CAR transgene in T cells, which is achieved by delivering transgene using integrating gamma retrovirus (RV) or lenti...

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Main Authors: Chuan Jin, Grammatiki Fotaki, Mohanraj Ramachandran, Berith Nilsson, Magnus Essand, Di Yu
Format: Article
Language:English
Published: Springer Nature 2016-05-01
Series:EMBO Molecular Medicine
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Online Access:https://doi.org/10.15252/emmm.201505869
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author Chuan Jin
Grammatiki Fotaki
Mohanraj Ramachandran
Berith Nilsson
Magnus Essand
Di Yu
author_facet Chuan Jin
Grammatiki Fotaki
Mohanraj Ramachandran
Berith Nilsson
Magnus Essand
Di Yu
author_sort Chuan Jin
collection DOAJ
description Abstract Chimeric antigen receptor (CAR) T‐cell therapy is a new successful treatment for refractory B‐cell leukemia. Successful therapeutic outcome depends on long‐term expression of CAR transgene in T cells, which is achieved by delivering transgene using integrating gamma retrovirus (RV) or lentivirus (LV). However, uncontrolled RV/LV integration in host cell genomes has the potential risk of causing insertional mutagenesis. Herein, we describe a novel episomal long‐term cell engineering method using non‐integrating lentiviral (NILV) vector containing a scaffold/matrix attachment region (S/MAR) element, for either expression of transgenes or silencing of target genes. The insertional events of this vector into the genome of host cells are below detection level. CD19 CAR T cells engineered with a NILV‐S/MAR vector have similar levels of CAR expression as T cells engineered with an integrating LV vector, even after numerous rounds of cell division. NILV‐S/MAR‐engineered CD19 CAR T cells exhibited similar cytotoxic capacity upon CD19+ target cell recognition as LV‐engineered T cells and are as effective in controlling tumor growth in vivo. We propose that NILV‐S/MAR vectors are superior to current options as they enable long‐term transgene expression without the risk of insertional mutagenesis and genotoxicity.
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issn 1757-4676
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publishDate 2016-05-01
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spelling doaj-art-baffc314e99e407f8134e568e1e6c5ca2025-08-20T03:06:00ZengSpringer NatureEMBO Molecular Medicine1757-46761757-46842016-05-018770271110.15252/emmm.201505869Safe engineering of CAR T cells for adoptive cell therapy of cancer using long‐term episomal gene transferChuan Jin0Grammatiki Fotaki1Mohanraj Ramachandran2Berith Nilsson3Magnus Essand4Di Yu5Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala UniversityDepartment of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala UniversityDepartment of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala UniversityDepartment of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala UniversityDepartment of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala UniversityDepartment of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala UniversityAbstract Chimeric antigen receptor (CAR) T‐cell therapy is a new successful treatment for refractory B‐cell leukemia. Successful therapeutic outcome depends on long‐term expression of CAR transgene in T cells, which is achieved by delivering transgene using integrating gamma retrovirus (RV) or lentivirus (LV). However, uncontrolled RV/LV integration in host cell genomes has the potential risk of causing insertional mutagenesis. Herein, we describe a novel episomal long‐term cell engineering method using non‐integrating lentiviral (NILV) vector containing a scaffold/matrix attachment region (S/MAR) element, for either expression of transgenes or silencing of target genes. The insertional events of this vector into the genome of host cells are below detection level. CD19 CAR T cells engineered with a NILV‐S/MAR vector have similar levels of CAR expression as T cells engineered with an integrating LV vector, even after numerous rounds of cell division. NILV‐S/MAR‐engineered CD19 CAR T cells exhibited similar cytotoxic capacity upon CD19+ target cell recognition as LV‐engineered T cells and are as effective in controlling tumor growth in vivo. We propose that NILV‐S/MAR vectors are superior to current options as they enable long‐term transgene expression without the risk of insertional mutagenesis and genotoxicity.https://doi.org/10.15252/emmm.201505869CAR T cellsepisomal cell engineeringnon‐integrating lentivirus (NILV)scaffold/matrix attachment region (S/MAR) elementself‐replicating DNA
spellingShingle Chuan Jin
Grammatiki Fotaki
Mohanraj Ramachandran
Berith Nilsson
Magnus Essand
Di Yu
Safe engineering of CAR T cells for adoptive cell therapy of cancer using long‐term episomal gene transfer
EMBO Molecular Medicine
CAR T cells
episomal cell engineering
non‐integrating lentivirus (NILV)
scaffold/matrix attachment region (S/MAR) element
self‐replicating DNA
title Safe engineering of CAR T cells for adoptive cell therapy of cancer using long‐term episomal gene transfer
title_full Safe engineering of CAR T cells for adoptive cell therapy of cancer using long‐term episomal gene transfer
title_fullStr Safe engineering of CAR T cells for adoptive cell therapy of cancer using long‐term episomal gene transfer
title_full_unstemmed Safe engineering of CAR T cells for adoptive cell therapy of cancer using long‐term episomal gene transfer
title_short Safe engineering of CAR T cells for adoptive cell therapy of cancer using long‐term episomal gene transfer
title_sort safe engineering of car t cells for adoptive cell therapy of cancer using long term episomal gene transfer
topic CAR T cells
episomal cell engineering
non‐integrating lentivirus (NILV)
scaffold/matrix attachment region (S/MAR) element
self‐replicating DNA
url https://doi.org/10.15252/emmm.201505869
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