Cargando…

Epstein-Barr viral product-containing exosomes promote fibrosis and nasopharyngeal carcinoma progression through activation of YAP1/FAPα signaling in fibroblasts

BACKGROUND: The progression of nasopharyngeal carcinoma (NPC) is profoundly affected by Epstein-Barr virus (EBV) infection. However, the role of EBV in the intercommunication between NPC and surrounding stromal cells has yet to be explored. METHODS: NPC biopsies were obtained for immunohistochemical...

Descripción completa

Detalles Bibliográficos
Autores principales: Lee, Po-Ju, Sui, Yun-Hua, Liu, Tzu-Tung, Tsang, Ngan-Ming, Huang, Chen-Han, Lin, Ting-Yi, Chang, Kai-Ping, Liu, Shu-Chen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9392321/
https://www.ncbi.nlm.nih.gov/pubmed/35986369
http://dx.doi.org/10.1186/s13046-022-02456-5
_version_ 1784771037002465280
author Lee, Po-Ju
Sui, Yun-Hua
Liu, Tzu-Tung
Tsang, Ngan-Ming
Huang, Chen-Han
Lin, Ting-Yi
Chang, Kai-Ping
Liu, Shu-Chen
author_facet Lee, Po-Ju
Sui, Yun-Hua
Liu, Tzu-Tung
Tsang, Ngan-Ming
Huang, Chen-Han
Lin, Ting-Yi
Chang, Kai-Ping
Liu, Shu-Chen
author_sort Lee, Po-Ju
collection PubMed
description BACKGROUND: The progression of nasopharyngeal carcinoma (NPC) is profoundly affected by Epstein-Barr virus (EBV) infection. However, the role of EBV in the intercommunication between NPC and surrounding stromal cells has yet to be explored. METHODS: NPC biopsies were obtained for immunohistochemical (IHC) analyses. Clinical correlations between the expression of active YAP1/FAPα and the fibrotic response and between YAP1/FAPα and the density of cytotoxic CD8a(+) T lymphocytes were determined. Survival times based on IHC scores were compared between groups using Kaplan-Meier survival and log-rank tests. Independent prognostic factors for metastasis/recurrence-free survival and overall survival were identified using univariate and multivariate Cox regression models. Fibroblasts were isolated from human nasopharyngeal biopsies. Exosomes were purified from culture supernatants of EBV(+)-positive NPC cells. The effects of EBV product-containing exosomes on fibroblast activation, fibrotic response, tumor growth, immune response, and correlations between the expression of featured genes were investigated using gel contraction assays, ELISAs, EdU incorporation assays, real-time impedance assays, RNA sequencing, immunostaining, 3D cancer spheroid coculture systems, and an NPC xenograft model. RESULTS: NPC patients who developed metastasis had significantly higher levels of active YAP1 and FAPα in their tumor stroma, which was further correlated with tumor fibrosis and poorer metastasis-free survival. Exosomes released from EBV(+)-NPC cells contained abundant FAPα protein and EBV-encoded latent membrane protein 1. Viral product-containing exosomes markedly enhanced the fibrotic response and tumor growth in a mouse xenograft model. IHC analyses of human NPC and NPC xenografts revealed positive correlations between levels of active YAP1 and FAPα, YAP1 and the fibrotic response, and FAPα and the fibrotic response. Mechanistic studies showed that treatment of fibroblasts with viral product-containing exosomes promoted the characteristics of cancer-associated fibroblasts by stimulating YAP1 signaling and the production of the immunosuppressive cytokines IL8, CCL2, and IL6. Inhibition of YAP1 activation markedly reversed these exosome-mediated protumoral effects, resulting in reduced contractility, inactivation of YAP1 signaling, and decreased production of immunosuppressive cytokines in fibroblasts. Furthermore, fibroblasts stimulated with these viral product-containing exosomes promoted NPC resistance to T cell-mediated cytotoxicity within tumor spheroids. In NPC tissues, a significant negative correlation was found between YAP1/FAPα and the density of CD8a(+) T lymphocytes with a granzyme B signature. CONCLUSION: EBV orchestrates interactions with the host and surrounding stroma by stimulating the functions of YAP1 and FAPα in fibroblasts through exosome cargos to create a more immunosuppressive, proinvasive microenvironment. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13046-022-02456-5.
format Online
Article
Text
id pubmed-9392321
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-93923212022-08-21 Epstein-Barr viral product-containing exosomes promote fibrosis and nasopharyngeal carcinoma progression through activation of YAP1/FAPα signaling in fibroblasts Lee, Po-Ju Sui, Yun-Hua Liu, Tzu-Tung Tsang, Ngan-Ming Huang, Chen-Han Lin, Ting-Yi Chang, Kai-Ping Liu, Shu-Chen J Exp Clin Cancer Res Research BACKGROUND: The progression of nasopharyngeal carcinoma (NPC) is profoundly affected by Epstein-Barr virus (EBV) infection. However, the role of EBV in the intercommunication between NPC and surrounding stromal cells has yet to be explored. METHODS: NPC biopsies were obtained for immunohistochemical (IHC) analyses. Clinical correlations between the expression of active YAP1/FAPα and the fibrotic response and between YAP1/FAPα and the density of cytotoxic CD8a(+) T lymphocytes were determined. Survival times based on IHC scores were compared between groups using Kaplan-Meier survival and log-rank tests. Independent prognostic factors for metastasis/recurrence-free survival and overall survival were identified using univariate and multivariate Cox regression models. Fibroblasts were isolated from human nasopharyngeal biopsies. Exosomes were purified from culture supernatants of EBV(+)-positive NPC cells. The effects of EBV product-containing exosomes on fibroblast activation, fibrotic response, tumor growth, immune response, and correlations between the expression of featured genes were investigated using gel contraction assays, ELISAs, EdU incorporation assays, real-time impedance assays, RNA sequencing, immunostaining, 3D cancer spheroid coculture systems, and an NPC xenograft model. RESULTS: NPC patients who developed metastasis had significantly higher levels of active YAP1 and FAPα in their tumor stroma, which was further correlated with tumor fibrosis and poorer metastasis-free survival. Exosomes released from EBV(+)-NPC cells contained abundant FAPα protein and EBV-encoded latent membrane protein 1. Viral product-containing exosomes markedly enhanced the fibrotic response and tumor growth in a mouse xenograft model. IHC analyses of human NPC and NPC xenografts revealed positive correlations between levels of active YAP1 and FAPα, YAP1 and the fibrotic response, and FAPα and the fibrotic response. Mechanistic studies showed that treatment of fibroblasts with viral product-containing exosomes promoted the characteristics of cancer-associated fibroblasts by stimulating YAP1 signaling and the production of the immunosuppressive cytokines IL8, CCL2, and IL6. Inhibition of YAP1 activation markedly reversed these exosome-mediated protumoral effects, resulting in reduced contractility, inactivation of YAP1 signaling, and decreased production of immunosuppressive cytokines in fibroblasts. Furthermore, fibroblasts stimulated with these viral product-containing exosomes promoted NPC resistance to T cell-mediated cytotoxicity within tumor spheroids. In NPC tissues, a significant negative correlation was found between YAP1/FAPα and the density of CD8a(+) T lymphocytes with a granzyme B signature. CONCLUSION: EBV orchestrates interactions with the host and surrounding stroma by stimulating the functions of YAP1 and FAPα in fibroblasts through exosome cargos to create a more immunosuppressive, proinvasive microenvironment. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13046-022-02456-5. BioMed Central 2022-08-20 /pmc/articles/PMC9392321/ /pubmed/35986369 http://dx.doi.org/10.1186/s13046-022-02456-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://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
Lee, Po-Ju
Sui, Yun-Hua
Liu, Tzu-Tung
Tsang, Ngan-Ming
Huang, Chen-Han
Lin, Ting-Yi
Chang, Kai-Ping
Liu, Shu-Chen
Epstein-Barr viral product-containing exosomes promote fibrosis and nasopharyngeal carcinoma progression through activation of YAP1/FAPα signaling in fibroblasts
title Epstein-Barr viral product-containing exosomes promote fibrosis and nasopharyngeal carcinoma progression through activation of YAP1/FAPα signaling in fibroblasts
title_full Epstein-Barr viral product-containing exosomes promote fibrosis and nasopharyngeal carcinoma progression through activation of YAP1/FAPα signaling in fibroblasts
title_fullStr Epstein-Barr viral product-containing exosomes promote fibrosis and nasopharyngeal carcinoma progression through activation of YAP1/FAPα signaling in fibroblasts
title_full_unstemmed Epstein-Barr viral product-containing exosomes promote fibrosis and nasopharyngeal carcinoma progression through activation of YAP1/FAPα signaling in fibroblasts
title_short Epstein-Barr viral product-containing exosomes promote fibrosis and nasopharyngeal carcinoma progression through activation of YAP1/FAPα signaling in fibroblasts
title_sort epstein-barr viral product-containing exosomes promote fibrosis and nasopharyngeal carcinoma progression through activation of yap1/fapα signaling in fibroblasts
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9392321/
https://www.ncbi.nlm.nih.gov/pubmed/35986369
http://dx.doi.org/10.1186/s13046-022-02456-5
work_keys_str_mv AT leepoju epsteinbarrviralproductcontainingexosomespromotefibrosisandnasopharyngealcarcinomaprogressionthroughactivationofyap1fapasignalinginfibroblasts
AT suiyunhua epsteinbarrviralproductcontainingexosomespromotefibrosisandnasopharyngealcarcinomaprogressionthroughactivationofyap1fapasignalinginfibroblasts
AT liutzutung epsteinbarrviralproductcontainingexosomespromotefibrosisandnasopharyngealcarcinomaprogressionthroughactivationofyap1fapasignalinginfibroblasts
AT tsangnganming epsteinbarrviralproductcontainingexosomespromotefibrosisandnasopharyngealcarcinomaprogressionthroughactivationofyap1fapasignalinginfibroblasts
AT huangchenhan epsteinbarrviralproductcontainingexosomespromotefibrosisandnasopharyngealcarcinomaprogressionthroughactivationofyap1fapasignalinginfibroblasts
AT lintingyi epsteinbarrviralproductcontainingexosomespromotefibrosisandnasopharyngealcarcinomaprogressionthroughactivationofyap1fapasignalinginfibroblasts
AT changkaiping epsteinbarrviralproductcontainingexosomespromotefibrosisandnasopharyngealcarcinomaprogressionthroughactivationofyap1fapasignalinginfibroblasts
AT liushuchen epsteinbarrviralproductcontainingexosomespromotefibrosisandnasopharyngealcarcinomaprogressionthroughactivationofyap1fapasignalinginfibroblasts