Organ-specific uptake of extracellular vesicles secreted by urological cancer cells

Extracellular vesicles (EVs) secreted by cancer cells have been shown to take a pivotal part in the process of local and systemic tumor progression by promoting the formation of a supportive local tumor microenvironment and preparing premetastatic niches in distant organ systems. In this study, we analyzed the organ-specific uptake of EVs secreted by urological cancer cells using an innovative in-vivo approach. EVs from benign and malignant prostate, kidney, and bladder cells were isolated using ultracentrifugation, fluorescence-labeled and injected intravenously in immunodeficient mice. After 12 or 24 h, the animals were sacrificed, their organs were harvested and analyzed for the presence of EVs by high-resolution fluorescence microscopy. Across all entities, EVs were taken up fast (12 h > 24 h), and EVs from malignant cells were taken up more efficiently than EVs from benign cells. Though not entirely organ-specific, EVs were incorporated in different amounts, depending on the entity (prostate: lung > liver > brain; kidney: brain > lung > liver; bladder: lung > liver > brain). EV uptake in other organs than lung, liver, brain, and spleen was not observed. Our results suggest a role of EVs in the formation of premetastatic niches and an organotropism in EV uptake, which have to be examined in more detail in further studies.

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Inhibition of a v b 3 integrin impairs adhesion and uptake of tumor-derived small extracellular vesicles

10.21203/rs.3.rs-17483/v1 ◽

2020 ◽

Author(s):

Wanessa Altei ◽

Bianca Pachane ◽

Patty K. Santos ◽

Ligia Ribeiro ◽

Bong Hwan Sung ◽

...

Keyword(s):

Cancer Cells ◽

Extracellular Vesicles ◽

Cell Communication ◽

Organ Specific ◽

Breast Cells ◽

Mediate Cell ◽

First Time ◽

Cd63 Expression ◽

Cell Cell

Abstract Background: Extracellular vesicles (EVs) are lipid-bound particles that are naturally released from cells and mediate cell-cell communication. Integrin adhesion receptors are enriched in small EVs (SEVs) and SEV-carried integrins have been shown to promote cancer cell migration and to mediate organ-specific metastasis; however, how integrins mediate these effects is not entirely clear and could represent a combination of EV binding to extracellular matrix and cells.Methods: To probe integrin role in EVs binding and uptake, we employed a disintegrin inhibitor (DisBa-01) of integrin binding with specificity for avb3 integrin. EVs were purified from MDA-MB-231 cells conditioned media by serial centrifugation method. Isolated EVs were characterized by different techniques and further employed in adhesion, uptake and co-culture experiments.Results: We find that SEVs secreted from MDA-MB-231 breast cancer cells carry avb3 integrin and bind directly to fibronectin-coated plates, which is inhibited by DisBa-01. SEV coating on tissue culture plates also induces adhesion of MDA-MB-231 cells, which is inhibited by DisBa-01 treatment. Analysis of EV uptake and interchange between cells reveals that the amount of CD63-positive EVs delivered from malignant MDA-MB-231 breast cells to non-malignant MCF10A breast epithelial cells is reduced by DisBa-01 treatment. Inhibition of avb3 integrin decreases CD63 expression in cancer cells suggesting an effect on SEV content.Conclusion: In summary, our findings demonstrate for the first time a key role of avb3 integrin in cell-cell communication through SEVs.

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Inhibition of a v b 3 integrin impairs adhesion and uptake of tumor-derived small extracellular vesicles

10.21203/rs.3.rs-17483/v2 ◽

2020 ◽

Author(s):

Wanessa Altei ◽

Bianca Pachane ◽

Patty K. Santos ◽

Ligia Ribeiro ◽

Bong Hwan Sung ◽

...

Keyword(s):

Cancer Cells ◽

Extracellular Vesicles ◽

Cell Communication ◽

Αvβ3 Integrin ◽

Organ Specific ◽

Breast Cells ◽

Mediate Cell ◽

First Time ◽

Cell Cell

Abstract Background: Extracellular vesicles (EVs) are lipid-bound particles that are naturally released from cells and mediate cell-cell communication. Integrin adhesion receptors are enriched in small EVs (SEVs) and SEV-carried integrins have been shown to promote cancer cell migration and to mediate organ-specific metastasis; however, how integrins mediate these effects is not entirely clear and could represent a combination of EV binding to extracellular matrix and cells. Methods: To probe integrin role in EVs binding and uptake, we employed a disintegrin inhibitor (DisBa-01) of integrin binding with specificity for αvβ3 integrin. EVs were purified from MDA-MB-231 cells conditioned media by serial centrifugation method. Isolated EVs were characterized by different techniques and further employed in adhesion, uptake and co-culture experiments. Results: We find that SEVs secreted from MDA-MB-231 breast cancer cells carry αvβ3 integrin and bind directly to fibronectin-coated plates, which is inhibited by DisBa-01. SEV coating on tissue culture plates also induces adhesion of MDA-MB-231 cells, which is inhibited by DisBa-01 treatment. Analysis of EV uptake and interchange between cells reveals that the amount of CD63-positive EVs delivered from malignant MDA-MB-231 breast cells to non-malignant MCF10A breast epithelial cells is reduced by DisBa-01 treatment. Inhibition of αvβ3 integrin decreases CD63 expression in cancer cells suggesting an effect on SEV content. Conclusion: In summary, our findings demonstrate for the first time a key role of αvβ3 integrin in cell-cell communication through SEVs.

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Inhibition of αvβ3 integrin impairs adhesion and uptake of tumor-derived small extracellular vesicles

Cell Communication and Signaling ◽

10.1186/s12964-020-00630-w ◽

2020 ◽

Vol 18 (1) ◽

Author(s):

Wanessa F. Altei ◽

Bianca C. Pachane ◽

Patty K. dos Santos ◽

Lígia N. M. Ribeiro ◽

Bong Hwan Sung ◽

...

Keyword(s):

Cancer Cells ◽

Extracellular Vesicles ◽

Cell Communication ◽

Αvβ3 Integrin ◽

Organ Specific ◽

Breast Cells ◽

Mediate Cell ◽

First Time ◽

Cell Cell

Abstract Background Extracellular vesicles (EVs) are lipid-bound particles that are naturally released from cells and mediate cell-cell communication. Integrin adhesion receptors are enriched in small EVs (SEVs) and SEV-carried integrins have been shown to promote cancer cell migration and to mediate organ-specific metastasis; however, how integrins mediate these effects is not entirely clear and could represent a combination of EV binding to extracellular matrix and cells. Methods To probe integrin role in EVs binding and uptake, we employed a disintegrin inhibitor (DisBa-01) of integrin binding with specificity for αvβ3 integrin. EVs were purified from MDA-MB-231 cells conditioned media by serial centrifugation method. Isolated EVs were characterized by different techniques and further employed in adhesion, uptake and co-culture experiments. Results We find that SEVs secreted from MDA-MB-231 breast cancer cells carry αvβ3 integrin and bind directly to fibronectin-coated plates, which is inhibited by DisBa-01. SEV coating on tissue culture plates also induces adhesion of MDA-MB-231 cells, which is inhibited by DisBa-01 treatment. Analysis of EV uptake and interchange between cells reveals that the amount of CD63-positive EVs delivered from malignant MDA-MB-231 breast cells to non-malignant MCF10A breast epithelial cells is reduced by DisBa-01 treatment. Inhibition of αvβ3 integrin decreases CD63 expression in cancer cells suggesting an effect on SEV content. Conclusion In summary, our findings demonstrate for the first time a key role of αvβ3 integrin in cell-cell communication through SEVs. Graphical abstract

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Organ-specific uptake of cancer-associated extracellular vesicles

European Urology Supplements ◽

10.1016/s1569-9056(19)33333-0 ◽

2019 ◽

Vol 18 (8) ◽

pp. e3086

Author(s):

J. Linxweiler ◽

A. Kolbinger ◽

M. Saar ◽

M. Stöckle ◽

K. Junker

Keyword(s):

Extracellular Vesicles ◽

Organ Specific ◽

Specific Uptake

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Study of NSCLC cell migration promoted by NSCLC-Derived Extracellular Vesicle using atomic force microscopy

Analytical Methods ◽

10.1039/d0ay02074e ◽

2021 ◽

Author(s):

Shuwei Wang ◽

Jiajia Wang ◽

Tuoyu Ju ◽

Kaige Qu ◽

Fan Yang ◽

...

Keyword(s):

Atomic Force Microscopy ◽

Cell Migration ◽

Cancer Cells ◽

Extracellular Vesicles ◽

Extracellular Vesicle ◽

Cancer Microenvironment ◽

Biological Processes ◽

Force Microscopy ◽

Atomic Force ◽

Cellular Components

Extracellular Vesicles (EVs) secreted by cancer cells have a key role in the cancer microenvironment and progression. Previous studies have mainly focused on molecular functions, cellular components and biological processes...

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Polymer-Coated Extracellular Vesicles for Selective Codelivery of Chemotherapeutics and siRNA to Cancer Cells

ACS Applied Bio Materials ◽

10.1021/acsabm.0c01153 ◽

2021 ◽

Vol 4 (2) ◽

pp. 1294-1306

Author(s):

Yong-Yu Jhan ◽

Guillermo Palou Zuniga ◽

Kanwar Abhay Singh ◽

Akhilesh K. Gaharwar ◽

Daniel L. Alge ◽

...

Keyword(s):

Cancer Cells ◽

Extracellular Vesicles

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Trastuzumab Modulates the Protein Cargo of Extracellular Vesicles Released by ERBB2+ Breast Cancer Cells

Membranes ◽

10.3390/membranes11030199 ◽

2021 ◽

Vol 11 (3) ◽

pp. 199

Author(s):

Silvia Marconi ◽

Sara Santamaria ◽

Martina Bartolucci ◽

Sara Stigliani ◽

Cinzia Aiello ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Extracellular Vesicles ◽

Breast Cancer Cells ◽

Culture Supernatant ◽

High Resolution Mass Spectrometry ◽

Recombinant Antibody ◽

Target Cells ◽

Cellular Processes ◽

Erbb2 Signaling

Cancers overexpressing the ERBB2 oncogene are aggressive and associated with a poor prognosis. Trastuzumab is an ERBB2 specific recombinant antibody employed for the treatment of these diseases since it blocks ERBB2 signaling causing growth arrest and survival inhibition. While the effects of Trastuzumab on ERBB2 cancer cells are well known, those on the extracellular vesicles (EVs) released from these cells are scarce. This study focused on ERBB2+ breast cancer cells and aimed to establish what type of EVs they release and whether Trastuzumab affects their morphology and molecular composition. To these aims, we performed immunoelectron microscopy, immunoblot, and high-resolution mass spectrometry analyses on EVs purified by differential centrifugation of culture supernatant. Here, we show that EVs released from ERBB2+ breast cancer cells are polymorphic in size and appearance and that ERBB2 is preferentially associated with large (120 nm) EVs. Moreover, we report that Trastuzumab (Tz) induces the expression of a specific glycosylated 50 kDa isoform of the CD63 tetraspanin and modulates the expression of 51 EVs proteins, including TOP1. Because these proteins are functionally associated with organelle organization, cytokinesis, and response to lipids, we suggest that Tz may influence these cellular processes in target cells at distant sites via modified EVs.

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