Isolation and analysis of tumor‑derived extracellular vesicles from head and neck squamous cell carcinoma plasma by galectin‑based glycan recognition particles

Benecke, Laura; Chiang, Dapi; Ebnoether, Eliane; Pfaffl, Michael W.; Müller, Laurent

doi:10.3892/ijo.2022.5423

Cited by 18 publications

(9 citation statements)

References 61 publications

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“…To verify the hypothesis of a stronger pro-angiogenic effect of TEVs within HNC patients’ sEVs, bead-based separation techniques of tumor-derived sEVs from plasma as described by others could be employed for future studies (Beccard et al 2020 ; Theodoraki et al 2018 ; Benecke et al 2022 ) and would provide a deeper insight into the angiogenic potential of TEVs and sEVs in general.…”

Section: Discussionmentioning

confidence: 99%

Plasma-derived small extracellular vesicles unleash the angiogenic potential in head and neck cancer patients

Tengler,

Schütz,

Tiedtke

et al. 2023

Mol Med

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Background In Head and neck cancer (HNC) angiogenesis is essential for tumor progression and metastasis. Small extracellular vesicles (sEVs) from HNC cell lines alter endothelial cell (EC) functions towards a pro-angiogenic phenotype. However, the role of plasma sEVs retrieved from HNC patients in this process is not clear so far. Methods Plasma sEVs were isolated on size exclusion chromatography columns from 32 HNC patients (early-stage UICC I/II: 8, advanced-stage UICC III/IV: 24), 12 patients with no evident disease after therapy (NED) and 16 healthy donors (HD). Briefly, sEVs were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), BCA protein assays and Western blots. Levels of angiogenesis-associated proteins were determined using antibody arrays. The interaction of fluorescently-labeled sEVs with human umbilical vein ECs was visualized by confocal microscopy. The functional effect of sEVs on tubulogenesis, migration, proliferation and apoptosis of ECs was assessed. Results The internalization of sEVs by ECs was visualized using confocal microscopy. Based on antibody arrays, all plasma sEVs were enriched in anti-angiogenic proteins. HNC sEVs contained more pro-angiogenic MMP-9 and anti-angiogenic proteins (Serpin F1) than HD sEVs. Interestingly, a strong inhibition of EC function was observed for sEVs from early-stage HNC, NED and HD. In contrast, sEVs from advanced-stage HNC showed a significantly increased tubulogenesis, migration and proliferation and induced less apoptosis in ECs than sEVs from HD. Conclusions In general, plasma sEVs carry a predominantly anti-angiogenic protein cargo and suppress the angiogenic properties of ECs, while sEVs from (advanced-stage) HNC patients induce angiogenesis compared to HD sEVs. Thus, tumor-derived sEVs within the plasma of HNC patients might shift the angiogenic switch towards angiogenesis.

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Section: Discussionmentioning

confidence: 99%

Plasma-derived small extracellular vesicles unleash the angiogenic potential in head and neck cancer patients

Tengler,

Schütz,

Tiedtke

et al. 2023

Mol Med

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“…Most commonly, the sizes of the membranous vesicles derived from tumor cells, including exosomes, range from 30 to 150 nm, while the size of the microvesicles ranges from 100 to 1000 nm [10][11][12]. In our study, the average size of isolated CIMVs from M14 cells was 240 nm, while CIMVs from M14-GM-CSF cells measured around 250 nm.…”

Section: Discussionmentioning

confidence: 44%

T-Lymphocytes Activated by Dendritic Cells Loaded by Tumor-Derived Vesicles Decrease Viability of Melanoma Cells In Vitro

Filin,

Mayasin,

Kharisova

et al. 2023

CIMB

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Immunotherapy represents an innovative approach to cancer treatment, based on activating the body’s own immune system to combat tumor cells. Among various immunotherapy strategies, dendritic cell vaccines hold a special place due to their ability to activate T-lymphocytes, key players in cellular immunity, and direct them to tumor cells. In this study, the influence of dendritic cells processed with tumor-derived vesicles on the viability of melanoma cells in vitro was investigated. Dendritic cells were loaded with tumor-derived vesicles, after which they were used to activate T-cells. The study demonstrated that such modified T-cells exhibit high activity against melanoma cells, leading to a decrease in their viability. Our analysis highlights the potential efficacy of this approach in developing immunotherapy against melanoma. These results provide new prospects for further research and the development of antitumor strategies based on the mechanisms of T-lymphocyte activation using tumor-derived vesicles.

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“…The cryo-EM sample preparation followed our previous publication ( Benecke et al, 2022 ). A total of 4 μL aliquot of each sample was adsorbed onto holey carbon-coated grid (Ted Pella Inc., Redding, United States) blotted with Whatman 1 filter paper and vitrified into liquid ethane at −178°C using a Leica GP plunger (Leica Microsystems GmbH, Wetzlar, Germany).…”

Section: Methodsmentioning

confidence: 99%

The proteome of bacterial membrane vesicles in Escherichia coli—a time course comparison study in two different media

Yu,

Chiang,

Reithmair

et al. 2024

Front. Microbiol.

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IntroductionBacteria inhabit the in- and outside of the human body, such as skin, gut or the oral cavity where they play an innoxious, beneficial or even pathogenic role. It is well known that bacteria can secrete membrane vesicles (MVs) like eukaryotic cells with extracellular vesicles (EVs). Several studies indicate that bacterial membrane vesicles (bMVs) play a crucial role in microbiome-host interactions. However, the composition of such bMVs and their functionality under different culture conditions are still largely unknown.MethodsTo gain a better insight into bMVs, we investigated the composition and functionality of E. coli (DSM 105380) bMVs from the culture media Lysogeny broth (LB) and RPMI 1640 throughout the different phases of growth (lag-, log- and stationary-phase). bMVs from three time points (8 h, 54 h, and 168 h) and two media (LB and RPMI 1640) were isolated by ultracentrifugation and analyzed using nanoparticle tracking analysis (NTA), cryogenic electron microscopy (Cryo-EM), conventional transmission electron microscopy (TEM) and mass spectrometry-based proteomics (LC–MS/MS). Furthermore, we examined pro-inflammatory cytokines IL-1β and IL-8 in the human monocyte cell line THP-1 upon bMV treatment.ResultsParticle numbers increased with inoculation periods. The bMV morphologies in Cryo-EM/TEM were similar at each time point and condition. Using proteomics, we identified 140 proteins, such as the common bMV markers OmpA and GroEL, present in bMVs isolated from both media and at all time points. Additionally, we were able to detect growth-condition-specific proteins. Treatment of THP-1 cells with bMVs of all six groups lead to significantly high IL-1β and IL-8 expressions.ConclusionOur study showed that the choice of medium and the duration of culturing significantly influence both E. coli bMV numbers and protein composition. Our TEM/Cryo-EM results demonstrated the presence of intact E. coli bMVs. Common E. coli proteins, including OmpA, GroEL, and ribosome proteins, can consistently be identified across all six tested growth conditions. Furthermore, our functional assays imply that bMVs isolated from the six groups retain their function and result in comparable cytokine induction.

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Isolation and analysis of tumor‑derived extracellular vesicles from head and neck squamous cell carcinoma plasma by galectin‑based glycan recognition particles

Cited by 18 publications

References 61 publications

Plasma-derived small extracellular vesicles unleash the angiogenic potential in head and neck cancer patients

Plasma-derived small extracellular vesicles unleash the angiogenic potential in head and neck cancer patients

T-Lymphocytes Activated by Dendritic Cells Loaded by Tumor-Derived Vesicles Decrease Viability of Melanoma Cells In Vitro

The proteome of bacterial membrane vesicles in Escherichia coli—a time course comparison study in two different media

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