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Interleukin-6-knockdown of chimeric antigen receptor-modified T cells significantly reduces IL-6 release from monocytes

BACKGROUND: T cells expressing a chimeric antigen receptor (CAR) engineered to target CD19 can treat leukemia effectively but also increase the risk of complications such as cytokine release syndrome (CRS) and CAR T cell related encephalopathy (CRES) driven by interleukin-6 (IL-6). Here, we investig...

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Autores principales: Kang, Liqing, Tang, Xiaowen, Zhang, Jian, Li, Minghao, Xu, Nan, Qi, Wei, Tan, Jingwen, Lou, Xiaoyan, Yu, Zhou, Sun, Juanjuan, Wang, Zhenkun, Dai, Haiping, Chen, Jia, Lin, Guoqing, Wu, Depei, Yu, Lei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7278071/
https://www.ncbi.nlm.nih.gov/pubmed/32523801
http://dx.doi.org/10.1186/s40164-020-00166-2
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author Kang, Liqing
Tang, Xiaowen
Zhang, Jian
Li, Minghao
Xu, Nan
Qi, Wei
Tan, Jingwen
Lou, Xiaoyan
Yu, Zhou
Sun, Juanjuan
Wang, Zhenkun
Dai, Haiping
Chen, Jia
Lin, Guoqing
Wu, Depei
Yu, Lei
author_facet Kang, Liqing
Tang, Xiaowen
Zhang, Jian
Li, Minghao
Xu, Nan
Qi, Wei
Tan, Jingwen
Lou, Xiaoyan
Yu, Zhou
Sun, Juanjuan
Wang, Zhenkun
Dai, Haiping
Chen, Jia
Lin, Guoqing
Wu, Depei
Yu, Lei
author_sort Kang, Liqing
collection PubMed
description BACKGROUND: T cells expressing a chimeric antigen receptor (CAR) engineered to target CD19 can treat leukemia effectively but also increase the risk of complications such as cytokine release syndrome (CRS) and CAR T cell related encephalopathy (CRES) driven by interleukin-6 (IL-6). Here, we investigated whether IL-6 knockdown in CART-19 cells can reduce IL-6 secretion from monocytes, which may reduce the risk of adverse events. METHODS: Supernatants from cocultures of regular CART-19 cells and B lymphoma cells were added to monocytes in vitro, and the IL-6 levels in monocyte supernatants were measured 24 h later. IL-6 expression was knocked down in regular CART-19 cells by adding a short hairpin RNA (shRNA) (termed ssCART-19) expression cassette specific for IL-6 to the conventional CAR vector. Transduction efficiency and cell proliferation were measured by flow cytometry, and cytotoxicity was measured by evaluating the release of lactate dehydrogenase into the medium. Gene expression was assessed by qRT-PCR and RNA sequencing. A xenograft leukemia mouse model was established by injecting NOD/SCID/γc-/- mice with luciferase-expressing B lymphoma cells, and then the animals were treated with regular CART-19 cells or ssCART-19. Tumor growth was assessed by bioluminescence imaging. RESULTS: Both recombinant IL-6 and CART-19 derived IL-6 significantly triggered IL-6 release by monocytes. IL-6 knockdown in ssCART-19 cells dramatically reduced IL-6 release from monocytes in vitro stduy. In vivo study further demonstrated that the mice bearing Raji cells treated with ssCART-19 cells showed significant lower IL-6 levels in serum than those treated with regular CART-19 cells, but comparable anti-tumor efficacy between the animal groups. CONCLUSION: CAR T-derived IL-6 is one of the most important initiators to amplify release of IL-6 from monocytes that further drive sCRS development. IL-6 knockdown in ssCART-19 cells by shRNA technology provide a promising strategy to improve the safety of CAR T cell therapy.
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spelling pubmed-72780712020-06-09 Interleukin-6-knockdown of chimeric antigen receptor-modified T cells significantly reduces IL-6 release from monocytes Kang, Liqing Tang, Xiaowen Zhang, Jian Li, Minghao Xu, Nan Qi, Wei Tan, Jingwen Lou, Xiaoyan Yu, Zhou Sun, Juanjuan Wang, Zhenkun Dai, Haiping Chen, Jia Lin, Guoqing Wu, Depei Yu, Lei Exp Hematol Oncol Research BACKGROUND: T cells expressing a chimeric antigen receptor (CAR) engineered to target CD19 can treat leukemia effectively but also increase the risk of complications such as cytokine release syndrome (CRS) and CAR T cell related encephalopathy (CRES) driven by interleukin-6 (IL-6). Here, we investigated whether IL-6 knockdown in CART-19 cells can reduce IL-6 secretion from monocytes, which may reduce the risk of adverse events. METHODS: Supernatants from cocultures of regular CART-19 cells and B lymphoma cells were added to monocytes in vitro, and the IL-6 levels in monocyte supernatants were measured 24 h later. IL-6 expression was knocked down in regular CART-19 cells by adding a short hairpin RNA (shRNA) (termed ssCART-19) expression cassette specific for IL-6 to the conventional CAR vector. Transduction efficiency and cell proliferation were measured by flow cytometry, and cytotoxicity was measured by evaluating the release of lactate dehydrogenase into the medium. Gene expression was assessed by qRT-PCR and RNA sequencing. A xenograft leukemia mouse model was established by injecting NOD/SCID/γc-/- mice with luciferase-expressing B lymphoma cells, and then the animals were treated with regular CART-19 cells or ssCART-19. Tumor growth was assessed by bioluminescence imaging. RESULTS: Both recombinant IL-6 and CART-19 derived IL-6 significantly triggered IL-6 release by monocytes. IL-6 knockdown in ssCART-19 cells dramatically reduced IL-6 release from monocytes in vitro stduy. In vivo study further demonstrated that the mice bearing Raji cells treated with ssCART-19 cells showed significant lower IL-6 levels in serum than those treated with regular CART-19 cells, but comparable anti-tumor efficacy between the animal groups. CONCLUSION: CAR T-derived IL-6 is one of the most important initiators to amplify release of IL-6 from monocytes that further drive sCRS development. IL-6 knockdown in ssCART-19 cells by shRNA technology provide a promising strategy to improve the safety of CAR T cell therapy. BioMed Central 2020-06-08 /pmc/articles/PMC7278071/ /pubmed/32523801 http://dx.doi.org/10.1186/s40164-020-00166-2 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Kang, Liqing
Tang, Xiaowen
Zhang, Jian
Li, Minghao
Xu, Nan
Qi, Wei
Tan, Jingwen
Lou, Xiaoyan
Yu, Zhou
Sun, Juanjuan
Wang, Zhenkun
Dai, Haiping
Chen, Jia
Lin, Guoqing
Wu, Depei
Yu, Lei
Interleukin-6-knockdown of chimeric antigen receptor-modified T cells significantly reduces IL-6 release from monocytes
title Interleukin-6-knockdown of chimeric antigen receptor-modified T cells significantly reduces IL-6 release from monocytes
title_full Interleukin-6-knockdown of chimeric antigen receptor-modified T cells significantly reduces IL-6 release from monocytes
title_fullStr Interleukin-6-knockdown of chimeric antigen receptor-modified T cells significantly reduces IL-6 release from monocytes
title_full_unstemmed Interleukin-6-knockdown of chimeric antigen receptor-modified T cells significantly reduces IL-6 release from monocytes
title_short Interleukin-6-knockdown of chimeric antigen receptor-modified T cells significantly reduces IL-6 release from monocytes
title_sort interleukin-6-knockdown of chimeric antigen receptor-modified t cells significantly reduces il-6 release from monocytes
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7278071/
https://www.ncbi.nlm.nih.gov/pubmed/32523801
http://dx.doi.org/10.1186/s40164-020-00166-2
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