Characterization of miRNAs in Extracellular Vesicles Released From Atlantic Salmon Monocyte-Like and Macrophage-Like Cells
Cell-derived extracellular vesicles (EVs) participate in cell-cell communication via transfer of molecular cargo including genetic material like miRNAs. In mammals, it has previously been established that EV-mediated transfer of miRNAs can alter the development or function of immune cells, such as m...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-11-01
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| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2020.587931/full |
| _version_ | 1818399349117288448 |
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| author | Nicole C. Smith Gabriel Wajnberg Simi Chacko Nardos T. Woldemariam Jacynthe Lacroix Nicolas Crapoulet D. Craig Ayre Stephen M. Lewis Stephen M. Lewis Stephen M. Lewis Matthew L. Rise Rune Andreassen Sherri L. Christian Sherri L. Christian |
| author_facet | Nicole C. Smith Gabriel Wajnberg Simi Chacko Nardos T. Woldemariam Jacynthe Lacroix Nicolas Crapoulet D. Craig Ayre Stephen M. Lewis Stephen M. Lewis Stephen M. Lewis Matthew L. Rise Rune Andreassen Sherri L. Christian Sherri L. Christian |
| author_sort | Nicole C. Smith |
| collection | DOAJ |
| description | Cell-derived extracellular vesicles (EVs) participate in cell-cell communication via transfer of molecular cargo including genetic material like miRNAs. In mammals, it has previously been established that EV-mediated transfer of miRNAs can alter the development or function of immune cells, such as macrophages. Our previous research revealed that Atlantic salmon head kidney leukocytes (HKLs) change their morphology, phagocytic ability and miRNA profile from primarily “monocyte-like” at Day 1 to primarily “macrophage-like” at Day 5 of culture. Therefore, we aimed to characterize the miRNA cargo packaged in EVs released from these two cell populations. We successfully isolated EVs from Atlantic salmon HKL culture supernatants using the established Vn96 peptide-based pull-down. Isolation was validated using transmission electron microscopy, nanoparticle tracking analysis, and Western blotting. RNA-sequencing identified 19 differentially enriched (DE) miRNAs packaged in Day 1 versus Day 5 EVs. Several of the highly abundant miRNAs, including those that were DE (e.g. ssa-miR-146a, ssa-miR-155 and ssa-miR-731), were previously identified as DE in HKLs and are associated with macrophage differentiation and immune response in other species. Interestingly, the abundance relative of the miRNAs in EVs, including the most abundant miRNA (ssa-miR-125b), was different than the miRNA abundance in HKLs, indicating selective packaging of miRNAs in EVs. Further study of the miRNA cargo in EVs derived from fish immune cells will be an important next step in identifying EV biomarkers useful for evaluating immune cell function, fish health, or response to disease. |
| first_indexed | 2024-12-14T07:19:16Z |
| format | Article |
| id | doaj.art-f758101d7eb444e9b24c1a159dab85da |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-14T07:19:16Z |
| publishDate | 2020-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-f758101d7eb444e9b24c1a159dab85da2022-12-21T23:11:39ZengFrontiers Media S.A.Frontiers in Immunology1664-32242020-11-011110.3389/fimmu.2020.587931587931Characterization of miRNAs in Extracellular Vesicles Released From Atlantic Salmon Monocyte-Like and Macrophage-Like CellsNicole C. Smith0Gabriel Wajnberg1Simi Chacko2Nardos T. Woldemariam3Jacynthe Lacroix4Nicolas Crapoulet5D. Craig Ayre6Stephen M. Lewis7Stephen M. Lewis8Stephen M. Lewis9Matthew L. Rise10Rune Andreassen11Sherri L. Christian12Sherri L. Christian13Department of Ocean Sciences, Memorial University, St. John’s, NL, Canada Atlantic Cancer Research Institute, Moncton, NB, Canada Atlantic Cancer Research Institute, Moncton, NB, CanadaDepartment of Life Sciences and Health, OsloMet-Oslo Metropolitan University, Oslo, Norway Atlantic Cancer Research Institute, Moncton, NB, Canada Atlantic Cancer Research Institute, Moncton, NB, CanadaDepartment of Molecular Sciences, University of Medicine and Health Sciences, Basseterre, Saint Kitts and Nevis Atlantic Cancer Research Institute, Moncton, NB, CanadaDepartment of Chemistry & Biochemistry, Université de Moncton, Moncton, NB, Canada Beatrice Hunter Cancer Research Institute, Halifax, NS, CanadaDepartment of Ocean Sciences, Memorial University, St. John’s, NL, CanadaDepartment of Life Sciences and Health, OsloMet-Oslo Metropolitan University, Oslo, Norway Beatrice Hunter Cancer Research Institute, Halifax, NS, CanadaDepartment of Biochemistry, Memorial University, St. John’s, NL, CanadaCell-derived extracellular vesicles (EVs) participate in cell-cell communication via transfer of molecular cargo including genetic material like miRNAs. In mammals, it has previously been established that EV-mediated transfer of miRNAs can alter the development or function of immune cells, such as macrophages. Our previous research revealed that Atlantic salmon head kidney leukocytes (HKLs) change their morphology, phagocytic ability and miRNA profile from primarily “monocyte-like” at Day 1 to primarily “macrophage-like” at Day 5 of culture. Therefore, we aimed to characterize the miRNA cargo packaged in EVs released from these two cell populations. We successfully isolated EVs from Atlantic salmon HKL culture supernatants using the established Vn96 peptide-based pull-down. Isolation was validated using transmission electron microscopy, nanoparticle tracking analysis, and Western blotting. RNA-sequencing identified 19 differentially enriched (DE) miRNAs packaged in Day 1 versus Day 5 EVs. Several of the highly abundant miRNAs, including those that were DE (e.g. ssa-miR-146a, ssa-miR-155 and ssa-miR-731), were previously identified as DE in HKLs and are associated with macrophage differentiation and immune response in other species. Interestingly, the abundance relative of the miRNAs in EVs, including the most abundant miRNA (ssa-miR-125b), was different than the miRNA abundance in HKLs, indicating selective packaging of miRNAs in EVs. Further study of the miRNA cargo in EVs derived from fish immune cells will be an important next step in identifying EV biomarkers useful for evaluating immune cell function, fish health, or response to disease.https://www.frontiersin.org/articles/10.3389/fimmu.2020.587931/fullextracellular vesiclesmicroRNAAtlantic salmonRNA-sequencingRNA-seqmacrophage |
| spellingShingle | Nicole C. Smith Gabriel Wajnberg Simi Chacko Nardos T. Woldemariam Jacynthe Lacroix Nicolas Crapoulet D. Craig Ayre Stephen M. Lewis Stephen M. Lewis Stephen M. Lewis Matthew L. Rise Rune Andreassen Sherri L. Christian Sherri L. Christian Characterization of miRNAs in Extracellular Vesicles Released From Atlantic Salmon Monocyte-Like and Macrophage-Like Cells Frontiers in Immunology extracellular vesicles microRNA Atlantic salmon RNA-sequencing RNA-seq macrophage |
| title | Characterization of miRNAs in Extracellular Vesicles Released From Atlantic Salmon Monocyte-Like and Macrophage-Like Cells |
| title_full | Characterization of miRNAs in Extracellular Vesicles Released From Atlantic Salmon Monocyte-Like and Macrophage-Like Cells |
| title_fullStr | Characterization of miRNAs in Extracellular Vesicles Released From Atlantic Salmon Monocyte-Like and Macrophage-Like Cells |
| title_full_unstemmed | Characterization of miRNAs in Extracellular Vesicles Released From Atlantic Salmon Monocyte-Like and Macrophage-Like Cells |
| title_short | Characterization of miRNAs in Extracellular Vesicles Released From Atlantic Salmon Monocyte-Like and Macrophage-Like Cells |
| title_sort | characterization of mirnas in extracellular vesicles released from atlantic salmon monocyte like and macrophage like cells |
| topic | extracellular vesicles microRNA Atlantic salmon RNA-sequencing RNA-seq macrophage |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2020.587931/full |
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