Extracellular Vesicles from Lung Adenocarcinoma Cells Induce Activation of Different Cancer-Associated Fibroblast Subtypes
<b>Background:</b> Lung cancer, including the major subtype lung adenocarcinoma (LUAD), is the leading cause of cancer deaths worldwide, largely due to metastasis. Improving survival rates requires new treatment strategies and a deeper understanding of the mechanisms that drive tumor pro...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-11-01
|
| Series: | Biomedicines |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2227-9059/12/11/2523 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850217192538243072 |
|---|---|
| author | Jessica Angelina Trejo Vazquez Rebecca Towle Dylan Andrew Farnsworth Masih Sarafan William Wallace Lockwood Cathie Garnis |
| author_facet | Jessica Angelina Trejo Vazquez Rebecca Towle Dylan Andrew Farnsworth Masih Sarafan William Wallace Lockwood Cathie Garnis |
| author_sort | Jessica Angelina Trejo Vazquez |
| collection | DOAJ |
| description | <b>Background:</b> Lung cancer, including the major subtype lung adenocarcinoma (LUAD), is the leading cause of cancer deaths worldwide, largely due to metastasis. Improving survival rates requires new treatment strategies and a deeper understanding of the mechanisms that drive tumor progression within the tumor microenvironment (TME). This study investigated the impact of extracellular vesicles (EVs) derived from LUAD cells on lung fibroblasts. <b>Methods:</b> EVs were isolated from LUAD cell lines via ultracentrifugation and characterized using nanoparticle tracking analysis and Western blotting. Lung fibroblasts were treated with PBS, TGFβ, or EVs, and their activation was assessed through protein (Western blotting) and RNA analyses (RNA seq and RT-qPCR). <b>Results:</b> The results confirmed the TGFβ induced activation and showed that LUAD EVs could also activate fibroblasts, increasing cancer-associated fibroblast (CAF) markers. While EV-induced CAF activation displayed unique features, like an increase in proliferation-related genes, the EV and TGFβ treatments also shared some differentially expressed genes. The EV groups induced a higher expression of ECM remodeling and EMT-associated genes, but some of those genes were also upregulated in the TGFβ group. Mesenchymal genes <i>POSTN</i> and <i>SPOCK1</i> were significantly upregulated in TGFβ- and EV-treated fibroblasts. Their secretion as proteins from the TGFβ- and EV-induced CAFs was not significant, confirmed through ELISA. <b>Conclusions:</b> These findings suggest that LUAD EVs play a role in CAF activation through both shared and distinct pathways compared to canonical TGFβ activation, potentially identifying novel gene expressions involved in CAF activation. Additionally, optimal protein secretion conditions of confirmed CAF-upregulated genes need to be established to determine their contribution to the TME. |
| format | Article |
| id | doaj-art-1288375b991a4dc08c17ab73fbc79e28 |
| institution | OA Journals |
| issn | 2227-9059 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomedicines |
| spelling | doaj-art-1288375b991a4dc08c17ab73fbc79e282025-08-20T02:08:08ZengMDPI AGBiomedicines2227-90592024-11-011211252310.3390/biomedicines12112523Extracellular Vesicles from Lung Adenocarcinoma Cells Induce Activation of Different Cancer-Associated Fibroblast SubtypesJessica Angelina Trejo Vazquez0Rebecca Towle1Dylan Andrew Farnsworth2Masih Sarafan3William Wallace Lockwood4Cathie Garnis5Department of Integrative Oncology, British Columbia Cancer Research Center, Vancouver, BC V5Z1L3, CanadaDepartment of Integrative Oncology, British Columbia Cancer Research Center, Vancouver, BC V5Z1L3, CanadaDepartment of Integrative Oncology, British Columbia Cancer Research Center, Vancouver, BC V5Z1L3, CanadaDepartment of Integrative Oncology, British Columbia Cancer Research Center, Vancouver, BC V5Z1L3, CanadaDepartment of Integrative Oncology, British Columbia Cancer Research Center, Vancouver, BC V5Z1L3, CanadaDepartment of Integrative Oncology, British Columbia Cancer Research Center, Vancouver, BC V5Z1L3, Canada<b>Background:</b> Lung cancer, including the major subtype lung adenocarcinoma (LUAD), is the leading cause of cancer deaths worldwide, largely due to metastasis. Improving survival rates requires new treatment strategies and a deeper understanding of the mechanisms that drive tumor progression within the tumor microenvironment (TME). This study investigated the impact of extracellular vesicles (EVs) derived from LUAD cells on lung fibroblasts. <b>Methods:</b> EVs were isolated from LUAD cell lines via ultracentrifugation and characterized using nanoparticle tracking analysis and Western blotting. Lung fibroblasts were treated with PBS, TGFβ, or EVs, and their activation was assessed through protein (Western blotting) and RNA analyses (RNA seq and RT-qPCR). <b>Results:</b> The results confirmed the TGFβ induced activation and showed that LUAD EVs could also activate fibroblasts, increasing cancer-associated fibroblast (CAF) markers. While EV-induced CAF activation displayed unique features, like an increase in proliferation-related genes, the EV and TGFβ treatments also shared some differentially expressed genes. The EV groups induced a higher expression of ECM remodeling and EMT-associated genes, but some of those genes were also upregulated in the TGFβ group. Mesenchymal genes <i>POSTN</i> and <i>SPOCK1</i> were significantly upregulated in TGFβ- and EV-treated fibroblasts. Their secretion as proteins from the TGFβ- and EV-induced CAFs was not significant, confirmed through ELISA. <b>Conclusions:</b> These findings suggest that LUAD EVs play a role in CAF activation through both shared and distinct pathways compared to canonical TGFβ activation, potentially identifying novel gene expressions involved in CAF activation. Additionally, optimal protein secretion conditions of confirmed CAF-upregulated genes need to be established to determine their contribution to the TME.https://www.mdpi.com/2227-9059/12/11/2523extracellular vesicleslung adenocarcinomacancer-associated fibroblaststumor microenvironmentcellular communication |
| spellingShingle | Jessica Angelina Trejo Vazquez Rebecca Towle Dylan Andrew Farnsworth Masih Sarafan William Wallace Lockwood Cathie Garnis Extracellular Vesicles from Lung Adenocarcinoma Cells Induce Activation of Different Cancer-Associated Fibroblast Subtypes Biomedicines extracellular vesicles lung adenocarcinoma cancer-associated fibroblasts tumor microenvironment cellular communication |
| title | Extracellular Vesicles from Lung Adenocarcinoma Cells Induce Activation of Different Cancer-Associated Fibroblast Subtypes |
| title_full | Extracellular Vesicles from Lung Adenocarcinoma Cells Induce Activation of Different Cancer-Associated Fibroblast Subtypes |
| title_fullStr | Extracellular Vesicles from Lung Adenocarcinoma Cells Induce Activation of Different Cancer-Associated Fibroblast Subtypes |
| title_full_unstemmed | Extracellular Vesicles from Lung Adenocarcinoma Cells Induce Activation of Different Cancer-Associated Fibroblast Subtypes |
| title_short | Extracellular Vesicles from Lung Adenocarcinoma Cells Induce Activation of Different Cancer-Associated Fibroblast Subtypes |
| title_sort | extracellular vesicles from lung adenocarcinoma cells induce activation of different cancer associated fibroblast subtypes |
| topic | extracellular vesicles lung adenocarcinoma cancer-associated fibroblasts tumor microenvironment cellular communication |
| url | https://www.mdpi.com/2227-9059/12/11/2523 |
| work_keys_str_mv | AT jessicaangelinatrejovazquez extracellularvesiclesfromlungadenocarcinomacellsinduceactivationofdifferentcancerassociatedfibroblastsubtypes AT rebeccatowle extracellularvesiclesfromlungadenocarcinomacellsinduceactivationofdifferentcancerassociatedfibroblastsubtypes AT dylanandrewfarnsworth extracellularvesiclesfromlungadenocarcinomacellsinduceactivationofdifferentcancerassociatedfibroblastsubtypes AT masihsarafan extracellularvesiclesfromlungadenocarcinomacellsinduceactivationofdifferentcancerassociatedfibroblastsubtypes AT williamwallacelockwood extracellularvesiclesfromlungadenocarcinomacellsinduceactivationofdifferentcancerassociatedfibroblastsubtypes AT cathiegarnis extracellularvesiclesfromlungadenocarcinomacellsinduceactivationofdifferentcancerassociatedfibroblastsubtypes |