Ciliary extracellular vesicles are distinct from the cytosolic extracellular vesicles
Abstract Extracellular vesicles (EVs) are cell‐derived membrane vesicles that are released into the extracellular space. EVs encapsulate key proteins and mediate intercellular signalling pathways. Recently, primary cilia have been shown to release EVs under fluid‐shear flow, but many proteins encaps...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2021-04-01
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| Series: | Journal of Extracellular Vesicles |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jev2.12086 |
| _version_ | 1818491414426681344 |
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| author | Ashraf M. Mohieldin Rajasekharreddy Pala Richard Beuttler James J. Moresco John R. Yates III Surya M. Nauli |
| author_facet | Ashraf M. Mohieldin Rajasekharreddy Pala Richard Beuttler James J. Moresco John R. Yates III Surya M. Nauli |
| author_sort | Ashraf M. Mohieldin |
| collection | DOAJ |
| description | Abstract Extracellular vesicles (EVs) are cell‐derived membrane vesicles that are released into the extracellular space. EVs encapsulate key proteins and mediate intercellular signalling pathways. Recently, primary cilia have been shown to release EVs under fluid‐shear flow, but many proteins encapsulated in these vesicles have never been identified. Primary cilia are ubiquitous mechanosensory organelles that protrude from the apical surface of almost all human cells. Primary cilia also serve as compartments for signalling pathways, and their defects have been associated with a wide range of human genetic diseases called ciliopathies. To better understand the mechanism of ciliopathies, it is imperative to know the distinctive protein profiles of the differently sourced EVs (cilia vs cytosol). Here, we isolated EVs from ciliated wild‐type (WT) and non‐ciliated IFT88 knockout (KO) mouse endothelial cells using fluid‐shear flow followed by a conventional method of EV isolation. EVs isolated from WT and KO exhibited distinctive sizes. Differences in EV protein contents were studied using liquid chromatography with tandem mass spectrometry (LC‐MS‐MS) and proteomic comparative analysis, which allowed us to classify proteins between ciliary EVs and cytosolic EVs derived from WT and KO, respectively. A total of 79 proteins were exclusively expressed in WT EVs, 145 solely in KO EVs, and 524 in both EVs. Our bioinformatics analyses revealed 29% distinct protein classes and 75% distinct signalling pathways between WT and KO EVs. Based on our statistical analyses and in vitro studies, we identified NADPH‐cytochrome P450 reductase (POR), and CD166 antigen (CD166) as potential biomarkers for ciliary and cytosolic EVs, respectively. Our protein‐protein interaction network analysis revealed that POR, but not CD166, interacted with either established or strong ciliopathy gene candidates. This report shows the unique differences between EVs secreted from cilia and the cytosol. These results will be important in advancing our understanding of human genetic diseases. |
| first_indexed | 2024-12-10T17:30:31Z |
| format | Article |
| id | doaj.art-36c900dac3da442991e7c15c934ccf65 |
| institution | Directory Open Access Journal |
| issn | 2001-3078 |
| language | English |
| last_indexed | 2024-12-10T17:30:31Z |
| publishDate | 2021-04-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Vesicles |
| spelling | doaj.art-36c900dac3da442991e7c15c934ccf652022-12-22T01:39:42ZengWileyJournal of Extracellular Vesicles2001-30782021-04-01106n/an/a10.1002/jev2.12086Ciliary extracellular vesicles are distinct from the cytosolic extracellular vesiclesAshraf M. Mohieldin0Rajasekharreddy Pala1Richard Beuttler2James J. Moresco3John R. Yates III4Surya M. Nauli5Department of Biomedical & Pharmaceutical Sciences Chapman University Irvine California USADepartment of Biomedical & Pharmaceutical Sciences Chapman University Irvine California USADepartment of Biomedical & Pharmaceutical Sciences Chapman University Irvine California USADepartment of Molecular Medicine The Scripps Research Institute La Jolla California USADepartment of Molecular Medicine The Scripps Research Institute La Jolla California USADepartment of Biomedical & Pharmaceutical Sciences Chapman University Irvine California USAAbstract Extracellular vesicles (EVs) are cell‐derived membrane vesicles that are released into the extracellular space. EVs encapsulate key proteins and mediate intercellular signalling pathways. Recently, primary cilia have been shown to release EVs under fluid‐shear flow, but many proteins encapsulated in these vesicles have never been identified. Primary cilia are ubiquitous mechanosensory organelles that protrude from the apical surface of almost all human cells. Primary cilia also serve as compartments for signalling pathways, and their defects have been associated with a wide range of human genetic diseases called ciliopathies. To better understand the mechanism of ciliopathies, it is imperative to know the distinctive protein profiles of the differently sourced EVs (cilia vs cytosol). Here, we isolated EVs from ciliated wild‐type (WT) and non‐ciliated IFT88 knockout (KO) mouse endothelial cells using fluid‐shear flow followed by a conventional method of EV isolation. EVs isolated from WT and KO exhibited distinctive sizes. Differences in EV protein contents were studied using liquid chromatography with tandem mass spectrometry (LC‐MS‐MS) and proteomic comparative analysis, which allowed us to classify proteins between ciliary EVs and cytosolic EVs derived from WT and KO, respectively. A total of 79 proteins were exclusively expressed in WT EVs, 145 solely in KO EVs, and 524 in both EVs. Our bioinformatics analyses revealed 29% distinct protein classes and 75% distinct signalling pathways between WT and KO EVs. Based on our statistical analyses and in vitro studies, we identified NADPH‐cytochrome P450 reductase (POR), and CD166 antigen (CD166) as potential biomarkers for ciliary and cytosolic EVs, respectively. Our protein‐protein interaction network analysis revealed that POR, but not CD166, interacted with either established or strong ciliopathy gene candidates. This report shows the unique differences between EVs secreted from cilia and the cytosol. These results will be important in advancing our understanding of human genetic diseases.https://doi.org/10.1002/jev2.12086bioinformaticsciliary ectosomesciliary exosomesciliary extracellular vesiclesciliary protein classesciliary vesicle proteome |
| spellingShingle | Ashraf M. Mohieldin Rajasekharreddy Pala Richard Beuttler James J. Moresco John R. Yates III Surya M. Nauli Ciliary extracellular vesicles are distinct from the cytosolic extracellular vesicles Journal of Extracellular Vesicles bioinformatics ciliary ectosomes ciliary exosomes ciliary extracellular vesicles ciliary protein classes ciliary vesicle proteome |
| title | Ciliary extracellular vesicles are distinct from the cytosolic extracellular vesicles |
| title_full | Ciliary extracellular vesicles are distinct from the cytosolic extracellular vesicles |
| title_fullStr | Ciliary extracellular vesicles are distinct from the cytosolic extracellular vesicles |
| title_full_unstemmed | Ciliary extracellular vesicles are distinct from the cytosolic extracellular vesicles |
| title_short | Ciliary extracellular vesicles are distinct from the cytosolic extracellular vesicles |
| title_sort | ciliary extracellular vesicles are distinct from the cytosolic extracellular vesicles |
| topic | bioinformatics ciliary ectosomes ciliary exosomes ciliary extracellular vesicles ciliary protein classes ciliary vesicle proteome |
| url | https://doi.org/10.1002/jev2.12086 |
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