Valosin-containing protein (VCP), a component of tumor-derived extracellular vesicles, impairs the barrier integrity of brain microvascular endothelial cells

Metastases are the leading cause of cancer-related deaths, and their origin is not fully elucidated. Recently, studies have shown that extracellular vesicles (EVs), particularly small extracellular vesicles (sEV), can disrupt the homeostasis of organs, promoting the development of a secondary tumor....

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Main Authors: Ramon Handerson Gomes Teles, Nicolas Jones Villarinho, Ana Sayuri Yamagata, Camila Tamy Hiroki, Murilo Camargo de Oliveira, Gisela Ramos Terçarioli, Ruy Gastaldoni Jaeger, Patrick Meybohm, Malgorzata Burek, Vanessa Morais Freitas
Format: Article
Language:English
Published: Elsevier 2025-01-01
Series:BBA Advances
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Online Access:http://www.sciencedirect.com/science/article/pii/S2667160324000188
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author Ramon Handerson Gomes Teles
Nicolas Jones Villarinho
Ana Sayuri Yamagata
Camila Tamy Hiroki
Murilo Camargo de Oliveira
Gisela Ramos Terçarioli
Ruy Gastaldoni Jaeger
Patrick Meybohm
Malgorzata Burek
Vanessa Morais Freitas
author_facet Ramon Handerson Gomes Teles
Nicolas Jones Villarinho
Ana Sayuri Yamagata
Camila Tamy Hiroki
Murilo Camargo de Oliveira
Gisela Ramos Terçarioli
Ruy Gastaldoni Jaeger
Patrick Meybohm
Malgorzata Burek
Vanessa Morais Freitas
author_sort Ramon Handerson Gomes Teles
collection DOAJ
description Metastases are the leading cause of cancer-related deaths, and their origin is not fully elucidated. Recently, studies have shown that extracellular vesicles (EVs), particularly small extracellular vesicles (sEV), can disrupt the homeostasis of organs, promoting the development of a secondary tumor. However, the role of sEV in brain endothelium and their association with metastasis related to breast cancer is unknown. Thus, this study aimed to investigate sEV-triggered changes in the phosphorylation state of proteins on the surface of brain endothelial cells, as they form the first barrier in contact with circulating tumor cells and EVs, and once identified, to modulate its interactors and effects from this through different functional assays. We used the most aggressive breast cancer cell line, MDA-MB-231, and its brain-seeking variant, MDA-MB-231-br. From these cells, small and large extracellular vesicles were harvested to treat hCMEC/D3 cells, an immortalized cell line from the human brain microvasculature. Higher levels of phosphorylation of VEGFR1 and VEGFR2 were found in hCMEC/D3 cells treated with MDA-MB-231-br sEV. By computational analysis, the Valosin-Containing Protein (VCP) was predicted to be an important sEV cargo affecting the VEGFR2 intracellular trafficking, validated by western blotting analysis. Then, VCP was modulated by cell transfection or chemical inhibition in hCMEC/D3 cells and assessed in different functional in vitro assays evidencing a significant effect on the functionality of these cells. Thus, this study demonstrates that the VCP-containing sEVs induce modifications at different phosphor sites of VEGFR2 and effectively modulate the state of brain microvascular endothelial cells.
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spelling doaj-art-7449669c8aa34a77afbb55f9bc87f8712025-08-20T01:58:16ZengElsevierBBA Advances2667-16032025-01-01710013010.1016/j.bbadva.2024.100130Valosin-containing protein (VCP), a component of tumor-derived extracellular vesicles, impairs the barrier integrity of brain microvascular endothelial cellsRamon Handerson Gomes Teles0Nicolas Jones Villarinho1Ana Sayuri Yamagata2Camila Tamy Hiroki3Murilo Camargo de Oliveira4Gisela Ramos Terçarioli5Ruy Gastaldoni Jaeger6Patrick Meybohm7Malgorzata Burek8Vanessa Morais Freitas9University of São Paulo, Department of Cell and Developmental Biology, Institute of Biomedical Sciences (ICB), São Paulo, 05508-000, Brazil; University Hospital Würzburg, Department of Anaesthesiology, Intensive Care, Emergency and Pain Medicine, 97080 Würzburg, Germany; University Würzburg, Graduate School of Life Sciences, Campus Hubland Nord, 97074 Würzburg, Germany; Corresponding author.University of São Paulo, Department of Cell and Developmental Biology, Institute of Biomedical Sciences (ICB), São Paulo, 05508-000, BrazilUniversity of São Paulo, Department of Cell and Developmental Biology, Institute of Biomedical Sciences (ICB), São Paulo, 05508-000, BrazilUniversity of São Paulo, Department of Cell and Developmental Biology, Institute of Biomedical Sciences (ICB), São Paulo, 05508-000, BrazilUniversity of São Paulo, Department of Cell and Developmental Biology, Institute of Biomedical Sciences (ICB), São Paulo, 05508-000, BrazilUniversity of São Paulo, Department of Cell and Developmental Biology, Institute of Biomedical Sciences (ICB), São Paulo, 05508-000, BrazilUniversity of São Paulo, Department of Cell and Developmental Biology, Institute of Biomedical Sciences (ICB), São Paulo, 05508-000, BrazilUniversity Hospital Würzburg, Department of Anaesthesiology, Intensive Care, Emergency and Pain Medicine, 97080 Würzburg, GermanyUniversity Hospital Würzburg, Department of Anaesthesiology, Intensive Care, Emergency and Pain Medicine, 97080 Würzburg, GermanyUniversity of São Paulo, Department of Cell and Developmental Biology, Institute of Biomedical Sciences (ICB), São Paulo, 05508-000, BrazilMetastases are the leading cause of cancer-related deaths, and their origin is not fully elucidated. Recently, studies have shown that extracellular vesicles (EVs), particularly small extracellular vesicles (sEV), can disrupt the homeostasis of organs, promoting the development of a secondary tumor. However, the role of sEV in brain endothelium and their association with metastasis related to breast cancer is unknown. Thus, this study aimed to investigate sEV-triggered changes in the phosphorylation state of proteins on the surface of brain endothelial cells, as they form the first barrier in contact with circulating tumor cells and EVs, and once identified, to modulate its interactors and effects from this through different functional assays. We used the most aggressive breast cancer cell line, MDA-MB-231, and its brain-seeking variant, MDA-MB-231-br. From these cells, small and large extracellular vesicles were harvested to treat hCMEC/D3 cells, an immortalized cell line from the human brain microvasculature. Higher levels of phosphorylation of VEGFR1 and VEGFR2 were found in hCMEC/D3 cells treated with MDA-MB-231-br sEV. By computational analysis, the Valosin-Containing Protein (VCP) was predicted to be an important sEV cargo affecting the VEGFR2 intracellular trafficking, validated by western blotting analysis. Then, VCP was modulated by cell transfection or chemical inhibition in hCMEC/D3 cells and assessed in different functional in vitro assays evidencing a significant effect on the functionality of these cells. Thus, this study demonstrates that the VCP-containing sEVs induce modifications at different phosphor sites of VEGFR2 and effectively modulate the state of brain microvascular endothelial cells.http://www.sciencedirect.com/science/article/pii/S2667160324000188Extracellular vesiclesExosomesMicrovesiclesBreast cancerMetastasishCMEC/D3
spellingShingle Ramon Handerson Gomes Teles
Nicolas Jones Villarinho
Ana Sayuri Yamagata
Camila Tamy Hiroki
Murilo Camargo de Oliveira
Gisela Ramos Terçarioli
Ruy Gastaldoni Jaeger
Patrick Meybohm
Malgorzata Burek
Vanessa Morais Freitas
Valosin-containing protein (VCP), a component of tumor-derived extracellular vesicles, impairs the barrier integrity of brain microvascular endothelial cells
BBA Advances
Extracellular vesicles
Exosomes
Microvesicles
Breast cancer
Metastasis
hCMEC/D3
title Valosin-containing protein (VCP), a component of tumor-derived extracellular vesicles, impairs the barrier integrity of brain microvascular endothelial cells
title_full Valosin-containing protein (VCP), a component of tumor-derived extracellular vesicles, impairs the barrier integrity of brain microvascular endothelial cells
title_fullStr Valosin-containing protein (VCP), a component of tumor-derived extracellular vesicles, impairs the barrier integrity of brain microvascular endothelial cells
title_full_unstemmed Valosin-containing protein (VCP), a component of tumor-derived extracellular vesicles, impairs the barrier integrity of brain microvascular endothelial cells
title_short Valosin-containing protein (VCP), a component of tumor-derived extracellular vesicles, impairs the barrier integrity of brain microvascular endothelial cells
title_sort valosin containing protein vcp a component of tumor derived extracellular vesicles impairs the barrier integrity of brain microvascular endothelial cells
topic Extracellular vesicles
Exosomes
Microvesicles
Breast cancer
Metastasis
hCMEC/D3
url http://www.sciencedirect.com/science/article/pii/S2667160324000188
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