Tetraspanins are unevenly distributed across single extracellular vesicles and bias sensitivity to multiplexed cancer biomarkers

Mizenko, Rachel R.; Brostoff, Terza; Rojalin, Tatu; Koster, Hanna J.; Swindell, Hila Shimshi; Leiserowitz, Gary S.; Wang, Aijun; Carney, Randy P.

doi:10.1186/s12951-021-00987-1

Cited by 95 publications

(75 citation statements)

References 58 publications

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“…Single-particle flow cytometry, such as image flow cytometry (IFCM), can analyze single sEV subpopulations by their surface protein profile, such as tetraspanins, at a high throughput rate [ 21 , 26 , 27 ]. However, it comes with a theoretical detection limit of around 80 nm for sEV [ 28 ].…”

Section: Introductionmentioning

confidence: 99%

Detailed Characterization of Small Extracellular Vesicles from Different Cell Types Based on Tetraspanin Composition by ExoView R100 Platform

Breitwieser

Koch

Tertel

et al. 2022

IJMS

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Small extracellular vesicles (sEV) hold enormous potential as biomarkers, drug carriers, and therapeutic agents. However, due to previous limitations in the phenotypic characterization of sEV at the single vesicle level, knowledge of cell type-specific sEV signatures remains sparse. With the introduction of next-generation sEV analysis devices, such as the single-particle interferometric reflectance imaging sensor (SP-IRIS)-based ExoView R100 platform, single sEV analyses are now possible. While the tetraspanins CD9, CD63, and CD81 were generally considered pan-sEV markers, it became clear that sEV of different cell types contain several combinations and amounts of these proteins on their surfaces. To gain better insight into the complexity and heterogeneity of sEV, we used the ExoView R100 platform to analyze the CD9/CD63/CD81 phenotype of sEV released by different cell types at a single sEV level. We demonstrated that these surface markers are sufficient to distinguish cell-type-specific sEV phenotypes. Furthermore, we recognized that tetraspanin composition in some sEV populations does not follow a random pattern. Notably, the tetraspanin distribution of sEV derived from mesenchymal stem cells (MSCs) alters depending on cell culture conditions. Overall, our data provide an overview of the cell-specific characteristics of sEV populations, which will increase the understanding of sEV physiology and improve the development of new sEV-based therapeutic approaches.

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Section: Introductionmentioning

confidence: 99%

Detailed Characterization of Small Extracellular Vesicles from Different Cell Types Based on Tetraspanin Composition by ExoView R100 Platform

Breitwieser

Koch

Tertel

et al. 2022

IJMS

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“…Good consistency exists in the tetraspanin profile of exosomes from the same source regardless of isolation methods. But the results confirmed that tetraspanin profiles of exosomes from a different source are distinct ( 134 ). Additionally, other exosomal cargos can also differ among different sources.…”

Section: Conclusion and Perspectivementioning

confidence: 52%

Potentiality of Exosomal Proteins as Novel Cancer Biomarkers for Liquid Biopsy

Jiang²,

Lv³

et al. 2022

Front. Immunol.

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Liquid biopsy has been rapidly developed in recent years due to its advantages of non-invasiveness and real-time sampling in cancer prognosis and diagnosis. Exosomes are nanosized extracellular vesicles secreted by all types of cells and abundantly distributed in all types of body fluid, carrying diverse cargos including proteins, DNA, and RNA, which transmit regulatory signals to recipient cells. Among the cargos, exosomal proteins have always been used as immunoaffinity binding targets for exosome isolation. Increasing evidence about the function of tumor-derived exosomes and their proteins is found to be massively associated with tumor initiation, progression, and metastasis in recent years. Therefore, exosomal proteins and some nucleic acids, such as miRNA, can be used not only as targets for exosome isolation but also as potential diagnostic markers in cancer research, especially for liquid biopsy. This review will discuss the existing protein-based methods for exosome isolation and characterization that are more appropriate for clinical use based on current knowledge of the exosomal biogenesis and function. Additionally, the recent studies for the use of exosomal proteins as cancer biomarkers are also discussed and summarized, which might contribute to the development of exosomal proteins as novel diagnostic tools for liquid biopsy.

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“…Until antibodies specific to equine EV-related proteins become available, we recommend using a combination of methods, including techniques that do not require antibodies, such as NTA and TEM. TEM can discriminate single EVs from similar-sized non-EV particles and is therefore widely used to monitor the quality and purity of EV-containing samples, e.g., for therapeutic application or downstream analysis [ 53 , 73 , 74 , 75 , 76 ].…”

Section: Discussionmentioning

confidence: 99%

Characterisation of Extracellular Vesicles from Equine Mesenchymal Stem Cells

Soukup¹,

Gerner

Gueltekin³

et al. 2022

IJMS

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Extracellular vesicles (EVs) are nanosized lipid bilayer-encapsulated particles secreted by virtually all cell types. EVs play an essential role in cellular crosstalk in health and disease. The cellular origin of EVs determines their composition and potential therapeutic effect. Mesenchymal stem/stromal cell (MSC)-derived EVs have shown a comparable therapeutic potential to their donor cells, making them a promising tool for regenerative medicine. The therapeutic application of EVs circumvents some safety concerns associated with the transplantation of viable, replicating cells and facilitates the quality-controlled production as a ready-to-go, off-the-shelf biological therapy. Recently, the International Society for Extracellular Vesicles (ISEV) suggested a set of minimal biochemical, biophysical and functional standards to define extracellular vesicles and their functions to improve standardisation in EV research. However, nonstandardised EV isolation methods and the limited availability of cross-reacting markers for most animal species restrict the application of these standards in the veterinary field and, therefore, the species comparability and standardisation of animal experiments. In this study, EVs were isolated from equine bone-marrow-derived MSCs using two different isolation methods, stepwise ultracentrifugation and size exclusion chromatography, and minimal experimental requirements for equine EVs were established and validated. Equine EVs were characterised using a nanotracking analysis, fluorescence-triggered flow cytometry, Western blot and transelectron microscopy. Based on the ISEV standards, minimal criteria for defining equine EVs are suggested as a baseline to allow the comparison of EV preparations obtained by different laboratories.

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Tetraspanins are unevenly distributed across single extracellular vesicles and bias sensitivity to multiplexed cancer biomarkers

Cited by 95 publications

References 58 publications

Detailed Characterization of Small Extracellular Vesicles from Different Cell Types Based on Tetraspanin Composition by ExoView R100 Platform

Detailed Characterization of Small Extracellular Vesicles from Different Cell Types Based on Tetraspanin Composition by ExoView R100 Platform

Potentiality of Exosomal Proteins as Novel Cancer Biomarkers for Liquid Biopsy

Characterisation of Extracellular Vesicles from Equine Mesenchymal Stem Cells

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