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MiRNA-340-5p mediates the functional and infiltrative promotion of tumor-infiltrating CD8(+) T lymphocytes in human diffuse large B cell lymphoma

BACKGROUND: CD8(+) tumor-infiltrating T lymphocytes (T-TILs) in the tumor microenvironment (TME) play an important role in tumor development, and miRNAs regulate tumor cell interactions with the microenvironment. T-TIL-based tumor immunotherapy provides a promising treatment strategy in diffuse larg...

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Autores principales: Xu, Yangyang, Liu, Zhenchuan, Lv, Lixin, Li, Ping, Xiu, Bing, Qian, Wenbin, Liang, Aibin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7653890/
https://www.ncbi.nlm.nih.gov/pubmed/33168024
http://dx.doi.org/10.1186/s13046-020-01752-2
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author Xu, Yangyang
Liu, Zhenchuan
Lv, Lixin
Li, Ping
Xiu, Bing
Qian, Wenbin
Liang, Aibin
author_facet Xu, Yangyang
Liu, Zhenchuan
Lv, Lixin
Li, Ping
Xiu, Bing
Qian, Wenbin
Liang, Aibin
author_sort Xu, Yangyang
collection PubMed
description BACKGROUND: CD8(+) tumor-infiltrating T lymphocytes (T-TILs) in the tumor microenvironment (TME) play an important role in tumor development, and miRNAs regulate tumor cell interactions with the microenvironment. T-TIL-based tumor immunotherapy provides a promising treatment strategy in diffuse large B-cell lymphoma (DLBCL). MiRNAs tend to be attractive targets for novel antitumor interventions. METHODS: Weighted gene coexpression network analysis (WGCNA), CIBERSORT analysis and Cox regression analysis were used to identify CD8(+) T-TIL-related miRNAs. RT-PCR, western blotting, immunohistochemistry (IHC), in situ hybridization (ISH), luciferase reporter assay, coimmunoprecipitation and ubiquitination analyses were used to detect miRNA, mRNA and protein expression and their combination. The viability and function of CD8(+) T cells after stimulation were evaluated by enzyme-linked immunosorbent assay (ELISA), cytotoxicity assay, functional avidity assessment, flow cytometry and Cell Counting Kit-8 (CCK-8) assay. DLBCL cell lines, primary cells and a murine xenograft model established with A20 cell injection were used as in vitro and in vivo experimental models. RESULTS: MiR-340-5p was positively correlated with CD8(+) T-TILs in DLBCL patients, and KMT5A was a direct target gene of miR-340-5p. CD8(+) T-cell function was significantly enhanced by miR-340-5p mimics both in vitro and in vivo, which was reversed by KMT5A overexpression. We demonstrated that COP1/CD73 was involved in the downstream mechanism of the miR-340-5p/KMT5A axis involving ubiquitination. In vivo, we validated an improved CD8(+) T-TIL infiltration rate and tumor suppression with miR-340-5p treatment. Furthermore, miR-340-5p directly regulated the biological activity of DLBCL cells without CD8(+) T-cell participation. CONCLUSIONS: MiR-340-5p promoted CD8(+) T-TIL infiltration and antitumor function by regulating KMT5A and COP1 and further activating CD73 ubiquitination. MiR-340-5p is potentially a novel target for DLBCL immunotherapy. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13046-020-01752-2.
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spelling pubmed-76538902020-11-10 MiRNA-340-5p mediates the functional and infiltrative promotion of tumor-infiltrating CD8(+) T lymphocytes in human diffuse large B cell lymphoma Xu, Yangyang Liu, Zhenchuan Lv, Lixin Li, Ping Xiu, Bing Qian, Wenbin Liang, Aibin J Exp Clin Cancer Res Research BACKGROUND: CD8(+) tumor-infiltrating T lymphocytes (T-TILs) in the tumor microenvironment (TME) play an important role in tumor development, and miRNAs regulate tumor cell interactions with the microenvironment. T-TIL-based tumor immunotherapy provides a promising treatment strategy in diffuse large B-cell lymphoma (DLBCL). MiRNAs tend to be attractive targets for novel antitumor interventions. METHODS: Weighted gene coexpression network analysis (WGCNA), CIBERSORT analysis and Cox regression analysis were used to identify CD8(+) T-TIL-related miRNAs. RT-PCR, western blotting, immunohistochemistry (IHC), in situ hybridization (ISH), luciferase reporter assay, coimmunoprecipitation and ubiquitination analyses were used to detect miRNA, mRNA and protein expression and their combination. The viability and function of CD8(+) T cells after stimulation were evaluated by enzyme-linked immunosorbent assay (ELISA), cytotoxicity assay, functional avidity assessment, flow cytometry and Cell Counting Kit-8 (CCK-8) assay. DLBCL cell lines, primary cells and a murine xenograft model established with A20 cell injection were used as in vitro and in vivo experimental models. RESULTS: MiR-340-5p was positively correlated with CD8(+) T-TILs in DLBCL patients, and KMT5A was a direct target gene of miR-340-5p. CD8(+) T-cell function was significantly enhanced by miR-340-5p mimics both in vitro and in vivo, which was reversed by KMT5A overexpression. We demonstrated that COP1/CD73 was involved in the downstream mechanism of the miR-340-5p/KMT5A axis involving ubiquitination. In vivo, we validated an improved CD8(+) T-TIL infiltration rate and tumor suppression with miR-340-5p treatment. Furthermore, miR-340-5p directly regulated the biological activity of DLBCL cells without CD8(+) T-cell participation. CONCLUSIONS: MiR-340-5p promoted CD8(+) T-TIL infiltration and antitumor function by regulating KMT5A and COP1 and further activating CD73 ubiquitination. MiR-340-5p is potentially a novel target for DLBCL immunotherapy. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13046-020-01752-2. BioMed Central 2020-11-10 /pmc/articles/PMC7653890/ /pubmed/33168024 http://dx.doi.org/10.1186/s13046-020-01752-2 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
Xu, Yangyang
Liu, Zhenchuan
Lv, Lixin
Li, Ping
Xiu, Bing
Qian, Wenbin
Liang, Aibin
MiRNA-340-5p mediates the functional and infiltrative promotion of tumor-infiltrating CD8(+) T lymphocytes in human diffuse large B cell lymphoma
title MiRNA-340-5p mediates the functional and infiltrative promotion of tumor-infiltrating CD8(+) T lymphocytes in human diffuse large B cell lymphoma
title_full MiRNA-340-5p mediates the functional and infiltrative promotion of tumor-infiltrating CD8(+) T lymphocytes in human diffuse large B cell lymphoma
title_fullStr MiRNA-340-5p mediates the functional and infiltrative promotion of tumor-infiltrating CD8(+) T lymphocytes in human diffuse large B cell lymphoma
title_full_unstemmed MiRNA-340-5p mediates the functional and infiltrative promotion of tumor-infiltrating CD8(+) T lymphocytes in human diffuse large B cell lymphoma
title_short MiRNA-340-5p mediates the functional and infiltrative promotion of tumor-infiltrating CD8(+) T lymphocytes in human diffuse large B cell lymphoma
title_sort mirna-340-5p mediates the functional and infiltrative promotion of tumor-infiltrating cd8(+) t lymphocytes in human diffuse large b cell lymphoma
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7653890/
https://www.ncbi.nlm.nih.gov/pubmed/33168024
http://dx.doi.org/10.1186/s13046-020-01752-2
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