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NOS1 inhibits the interferon response of cancer cells by S-nitrosylation of HDAC2

BACKGROUND: The dysfunction of type I interferon (IFN) signaling is an important mechanism of immune escape and metastasis in tumors. Increased NOS1 expression has been detected in melanoma, which correlated with dysfunctional IFN signaling and poor response to immunotherapy, but the specific mechan...

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Autores principales: Xu, Pengfei, Ye, Shuangyan, Li, Keyi, Huang, Mengqiu, Wang, Qianli, Zeng, Sisi, Chen, Xi, Gao, Wenwen, Chen, Jianping, Zhang, Qianbing, Zhong, Zhuo, Lin, Ying, Rong, Zhili, Xu, Yang, Hao, Bingtao, Peng, Anghui, Ouyang, Manzhao, Liu, Qiuzhen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6896289/
https://www.ncbi.nlm.nih.gov/pubmed/31805977
http://dx.doi.org/10.1186/s13046-019-1448-9
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author Xu, Pengfei
Ye, Shuangyan
Li, Keyi
Huang, Mengqiu
Wang, Qianli
Zeng, Sisi
Chen, Xi
Gao, Wenwen
Chen, Jianping
Zhang, Qianbing
Zhong, Zhuo
Lin, Ying
Rong, Zhili
Xu, Yang
Hao, Bingtao
Peng, Anghui
Ouyang, Manzhao
Liu, Qiuzhen
author_facet Xu, Pengfei
Ye, Shuangyan
Li, Keyi
Huang, Mengqiu
Wang, Qianli
Zeng, Sisi
Chen, Xi
Gao, Wenwen
Chen, Jianping
Zhang, Qianbing
Zhong, Zhuo
Lin, Ying
Rong, Zhili
Xu, Yang
Hao, Bingtao
Peng, Anghui
Ouyang, Manzhao
Liu, Qiuzhen
author_sort Xu, Pengfei
collection PubMed
description BACKGROUND: The dysfunction of type I interferon (IFN) signaling is an important mechanism of immune escape and metastasis in tumors. Increased NOS1 expression has been detected in melanoma, which correlated with dysfunctional IFN signaling and poor response to immunotherapy, but the specific mechanism has not been determined. In this study, we investigated the regulation of NOS1 on the interferon response and clarified the relevant molecular mechanisms. METHODS: After stable transfection of A375 cells with NOS1 expression plasmids, the transcription and expression of IFNα-stimulated genes (ISGs) were assessed using pISRE luciferase reporter gene analysis, RT-PCR, and western blotting, respectively. The effect of NOS1 on lung metastasis was assessed in melanoma mouse models. A biotin-switch assay was performed to detect the S-nitrosylation of HDAC2 by NOS1. ChIP-qPCR was conducted to measure the binding of HDAC2, H4K16ac, H4K5ac, H3ac, and RNA polymerase II in the promoters of ISGs after IFNα stimulation. This effect was further evaluated by altering the expression level of HDAC2 or by transfecting the HDAC2-C262A/C274A site mutant plasmids into cells. The coimmunoprecipitation assay was performed to detect the interaction of HDAC2 with STAT1 and STAT2. Loss-of-function and gain-of-function approaches were used to examine the effect of HDAC2-C262A/C274A on lung metastasis. Tumor infiltrating lymphocytes were analyzed by flow cytometry. RESULTS: HDAC2 is recruited to the promoter of ISGs and deacetylates H4K16 for the optimal expression of ISGs in response to IFNα treatment. Overexpression of NOS1 in melanoma cells decreases IFNα-responsiveness and induces the S-nitrosylation of HDAC2-C262/C274. This modification decreases the binding of HDAC2 with STAT1, thereby reducing the recruitment of HDAC2 to the ISG promoter and the deacetylation of H4K16. Moreover, expression of a mutant form of HDAC2, which cannot be nitrosylated, reverses the inhibition of ISG expression by NOS1 in vitro and decreases NOS1-induced lung metastasis and inhibition of tumor infiltrating lymphocytes in a melanoma mouse model. CONCLUSIONS: This study provides evidence that NOS1 induces dysfunctional IFN signaling to promote lung metastasis in melanoma, highlighting NOS1-induced S-nitrosylation of HDAC2 in the regulation of IFN signaling via histone modification.
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spelling pubmed-68962892019-12-11 NOS1 inhibits the interferon response of cancer cells by S-nitrosylation of HDAC2 Xu, Pengfei Ye, Shuangyan Li, Keyi Huang, Mengqiu Wang, Qianli Zeng, Sisi Chen, Xi Gao, Wenwen Chen, Jianping Zhang, Qianbing Zhong, Zhuo Lin, Ying Rong, Zhili Xu, Yang Hao, Bingtao Peng, Anghui Ouyang, Manzhao Liu, Qiuzhen J Exp Clin Cancer Res Research BACKGROUND: The dysfunction of type I interferon (IFN) signaling is an important mechanism of immune escape and metastasis in tumors. Increased NOS1 expression has been detected in melanoma, which correlated with dysfunctional IFN signaling and poor response to immunotherapy, but the specific mechanism has not been determined. In this study, we investigated the regulation of NOS1 on the interferon response and clarified the relevant molecular mechanisms. METHODS: After stable transfection of A375 cells with NOS1 expression plasmids, the transcription and expression of IFNα-stimulated genes (ISGs) were assessed using pISRE luciferase reporter gene analysis, RT-PCR, and western blotting, respectively. The effect of NOS1 on lung metastasis was assessed in melanoma mouse models. A biotin-switch assay was performed to detect the S-nitrosylation of HDAC2 by NOS1. ChIP-qPCR was conducted to measure the binding of HDAC2, H4K16ac, H4K5ac, H3ac, and RNA polymerase II in the promoters of ISGs after IFNα stimulation. This effect was further evaluated by altering the expression level of HDAC2 or by transfecting the HDAC2-C262A/C274A site mutant plasmids into cells. The coimmunoprecipitation assay was performed to detect the interaction of HDAC2 with STAT1 and STAT2. Loss-of-function and gain-of-function approaches were used to examine the effect of HDAC2-C262A/C274A on lung metastasis. Tumor infiltrating lymphocytes were analyzed by flow cytometry. RESULTS: HDAC2 is recruited to the promoter of ISGs and deacetylates H4K16 for the optimal expression of ISGs in response to IFNα treatment. Overexpression of NOS1 in melanoma cells decreases IFNα-responsiveness and induces the S-nitrosylation of HDAC2-C262/C274. This modification decreases the binding of HDAC2 with STAT1, thereby reducing the recruitment of HDAC2 to the ISG promoter and the deacetylation of H4K16. Moreover, expression of a mutant form of HDAC2, which cannot be nitrosylated, reverses the inhibition of ISG expression by NOS1 in vitro and decreases NOS1-induced lung metastasis and inhibition of tumor infiltrating lymphocytes in a melanoma mouse model. CONCLUSIONS: This study provides evidence that NOS1 induces dysfunctional IFN signaling to promote lung metastasis in melanoma, highlighting NOS1-induced S-nitrosylation of HDAC2 in the regulation of IFN signaling via histone modification. BioMed Central 2019-12-05 /pmc/articles/PMC6896289/ /pubmed/31805977 http://dx.doi.org/10.1186/s13046-019-1448-9 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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.
spellingShingle Research
Xu, Pengfei
Ye, Shuangyan
Li, Keyi
Huang, Mengqiu
Wang, Qianli
Zeng, Sisi
Chen, Xi
Gao, Wenwen
Chen, Jianping
Zhang, Qianbing
Zhong, Zhuo
Lin, Ying
Rong, Zhili
Xu, Yang
Hao, Bingtao
Peng, Anghui
Ouyang, Manzhao
Liu, Qiuzhen
NOS1 inhibits the interferon response of cancer cells by S-nitrosylation of HDAC2
title NOS1 inhibits the interferon response of cancer cells by S-nitrosylation of HDAC2
title_full NOS1 inhibits the interferon response of cancer cells by S-nitrosylation of HDAC2
title_fullStr NOS1 inhibits the interferon response of cancer cells by S-nitrosylation of HDAC2
title_full_unstemmed NOS1 inhibits the interferon response of cancer cells by S-nitrosylation of HDAC2
title_short NOS1 inhibits the interferon response of cancer cells by S-nitrosylation of HDAC2
title_sort nos1 inhibits the interferon response of cancer cells by s-nitrosylation of hdac2
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6896289/
https://www.ncbi.nlm.nih.gov/pubmed/31805977
http://dx.doi.org/10.1186/s13046-019-1448-9
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