Caveolin-1-Dependent Tenascin C Inclusion in Extracellular Vesicles Is Required to Promote Breast Cancer Cell Malignancy

Abstract

Background: Elevated expression of CAV1 in breast cancer increases tumor progression. Extracellular vesicles (EVs) from CAV1-expressing MDA-MB-231 breast cancer cells contain Tenascin C (TNC), but the relevance of TNC remained to be defined. Methods: EVs were characterized by nanotracking analysis, microscopy and western blotting. The uptake of EVs by cells was studied using flow cytometry. The effects of EVs on breast cancer cells were tested in migration, invasion, colony formation and in vivo assays. Results: EVs were taken up by cells; however, only those containing TNC promoted invasiveness. In vivo, EVs lacking TNC ceased to promote tumor growth. Conclusion: CAV1 and TNC contained in breast cancer cell-derived EVs were identified as proteins that favor progression of breast cancer.

Plain language summary

Caveolin-1 (CAV1) is a protein that in breast cancer increases with disease progression. Extracellular vesicles (EVs) from breast cancer cells with CAV1 also contain Tenascin C (TNC) protein, but the importance of TNC remained to be defined. EVs were identified by size, microscopy and protein analysis. The effects of EVs on breast cancer cells were studied using cells and experiments in animals. CAV1 expression promotes TNC inclusion into EVs, which increased the aggressiveness of recipient breast cancer cells. In animals, only EVs with TNC increased features associated with cancer spread, while EVs lacking TNC reduced tumor growth.

Keywords::

• breast cancer
• CAV1
• exosomes
• metastasis
• TNC

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.tandfonline.com/doi/suppl/10.2217/nnm-2023-0143

Author contributions

A Campos, R Burgos-Ravanal, L Lobos-Gonzalez and A Quest designed the study; A Campos, R Burgos-Ravanal, R Huilcaman, MF Gonzalez, J Diaz, A Caceres Verschae, JP Acevedo, M Carrasco, F Silva, E Jeldes, M Varas-Godoy and L Lobos-Gonzalzez performed experiments; L Leyton, L Lobos-Gonzalez and A Quest supervised the study; A Campos and R Burgos-Ravanal wrote the manuscript; L Leyton, L Lobos-Gonzalez and A Quest reviewed the manuscript. All authors contributed to the article and approved the submitted version.

Financial disclosure

This work was supported by FONDECYT grants 1210644 (AFGQ), 1200836 (LL), 1211223 (LL-G), 1190928 (MVG), FONDAP grant 15130011 (AFGQ, LL, LL-G, MVG), ANID/BASAL/FB210008 (MVG), ANID postdoctoral fellowship award Becas Chile (AC), 3170169 (JD), ANID PhD fellowship awards 21130102 (AC), 21200147 (RB-R), 21161246 (RH), 21170292 (MFG) and IMPACT, #FB210024 (JPA). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Competing interests disclosure

The authors have no competing interests or relevant affiliations with any organization or entity with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Writing disclosure

No writing assistance was utilized in the production of this manuscript.

Data availability

All pertinent data to support this study are included in the paper and supplementary files. Further data supporting the findings are available upon request.

Acknowledgments

This work was possible thanks to the use of the Nanosight NS300 equipment (FONDEQUIP EQM160157). The authors thank C Zuñiga and H Camus of the CEMC facility for their technical support relating to use of the ultracentrifuge (Sorvall WX + 100) and the fluorescence microscope (Spinning disk Olympus IX81). The authors also thank S Valenzuela and M Ricca for their valuable veterinary assistance in our animal facility. The authors also thank L Leyton for the use of her paid account to make the figures created with BioRender.com accessed on 6 November 2021.

Additional information

Funding

This work was supported by FONDECYT grants 1210644 (AFGQ), 1200836 (LL), 1211223 (LL-G), 1190928 (MVG), FONDAP grant 15130011 (AFGQ, LL, LL-G, MVG), ANID/BASAL/FB210008 (MVG), ANID postdoctoral fellowship award Becas Chile (AC), 3170169 (JD), ANID PhD fellowship awards 21130102 (AC), 21200147 (RB-R), 21161246 (RH), 21170292 (MFG) and IMPACT, #FB210024 (JPA). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.