SARS CoV-2 spike protein-guided exosome isolation facilitates detection of potential miRNA biomarkers in COVID-19 infections
© 2023 the author(s), published by De Gruyter, Berlin/Boston..
OBJECTIVES: Nearly three years into the pandemic, SARS-CoV-2 infections are occurring in vaccinated and naturally infected populations. While humoral and cellular responses in COVID-19 are being characterized, novel immune biomarkers also being identified. Recently, an increase in angiotensin-converting enzyme 2 expressing (aka, ACE2 positive) circulating exosomes (ExoACE2) were identified in the plasma of COVID-19 patients (El-Shennawy et al.). In this pilot study, we describe a method to characterize the exosome-associated microRNA (exo-miRNA) signature in ACE2-positive and ACE2-negative exosomal populations (non-ExoACE2).
METHODS: We performed a sorting protocol using the recombinant biotin-conjugated SARS CoV-2 spike protein containing the receptor binding domain (RBD) on plasma samples from six patients. Following purification, exo-miRNA were characterized for ACE2-positive and ACE2-negative exosome subpopulations by RT-PCR.
RESULTS: We identified differential expression of several miRNA. Specifically let-7g-5p and hsa-miR-4454+miR-7975 were upregulated, while hsa-miR-208a-3p and has-miR-323-3p were downregulated in ExoACE2 vs. non-ExoACE2.
CONCLUSIONS: The SARS CoV-2 spike-protein guided exosome isolation permits isolation of ExoACE2 exosomes. Such purification facilitates detailed characterization of potential biomarkers (e.g. exo-miRNA) for COVID-19 patients. This method could be used for future studies to further the understanding mechanisms of host response against SARS CoV-2.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:61 |
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| Enthalten in: |
Clinical chemistry and laboratory medicine - 61(2023), 8 vom: 26. Juli, Seite 1518-1524 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Mimmi, Selena [VerfasserIn] |
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| Links: |
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| Themen: |
ACE2 |
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| Anmerkungen: |
Date Completed 03.07.2023 Date Revised 21.11.2023 published: Electronic-Print Citation Status MEDLINE |
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| doi: |
10.1515/cclm-2022-1286 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM354777114 |
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| 245 | 1 | 0 | |a SARS CoV-2 spike protein-guided exosome isolation facilitates detection of potential miRNA biomarkers in COVID-19 infections |
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| 500 | |a Date Completed 03.07.2023 | ||
| 500 | |a Date Revised 21.11.2023 | ||
| 500 | |a published: Electronic-Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2023 the author(s), published by De Gruyter, Berlin/Boston. | ||
| 520 | |a OBJECTIVES: Nearly three years into the pandemic, SARS-CoV-2 infections are occurring in vaccinated and naturally infected populations. While humoral and cellular responses in COVID-19 are being characterized, novel immune biomarkers also being identified. Recently, an increase in angiotensin-converting enzyme 2 expressing (aka, ACE2 positive) circulating exosomes (ExoACE2) were identified in the plasma of COVID-19 patients (El-Shennawy et al.). In this pilot study, we describe a method to characterize the exosome-associated microRNA (exo-miRNA) signature in ACE2-positive and ACE2-negative exosomal populations (non-ExoACE2) | ||
| 520 | |a METHODS: We performed a sorting protocol using the recombinant biotin-conjugated SARS CoV-2 spike protein containing the receptor binding domain (RBD) on plasma samples from six patients. Following purification, exo-miRNA were characterized for ACE2-positive and ACE2-negative exosome subpopulations by RT-PCR | ||
| 520 | |a RESULTS: We identified differential expression of several miRNA. Specifically let-7g-5p and hsa-miR-4454+miR-7975 were upregulated, while hsa-miR-208a-3p and has-miR-323-3p were downregulated in ExoACE2 vs. non-ExoACE2 | ||
| 520 | |a CONCLUSIONS: The SARS CoV-2 spike-protein guided exosome isolation permits isolation of ExoACE2 exosomes. Such purification facilitates detailed characterization of potential biomarkers (e.g. exo-miRNA) for COVID-19 patients. This method could be used for future studies to further the understanding mechanisms of host response against SARS CoV-2 | ||
| 650 | 4 | |a Journal Article | |
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| 700 | 1 | |a Zimbo, Anna Maria |e verfasserin |4 aut | |
| 700 | 1 | |a Rotundo, Salvatore |e verfasserin |4 aut | |
| 700 | 1 | |a Cione, Erika |e verfasserin |4 aut | |
| 700 | 1 | |a Nisticò, Nancy |e verfasserin |4 aut | |
| 700 | 1 | |a Aloisio, Annamaria |e verfasserin |4 aut | |
| 700 | 1 | |a Maisano, Domenico |e verfasserin |4 aut | |
| 700 | 1 | |a Tolomeo, Anna Maria |e verfasserin |4 aut | |
| 700 | 1 | |a Dattilo, Vincenzo |e verfasserin |4 aut | |
| 700 | 1 | |a Lionello, Rosaria |e verfasserin |4 aut | |
| 700 | 1 | |a Fioravanti, Antonella |e verfasserin |4 aut | |
| 700 | 1 | |a Di Loria, Antonio |e verfasserin |4 aut | |
| 700 | 1 | |a Quirino, Angela |e verfasserin |4 aut | |
| 700 | 1 | |a Marascio, Nadia |e verfasserin |4 aut | |
| 700 | 1 | |a Russo, Alessandro |e verfasserin |4 aut | |
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| 700 | 1 | |a Torti, Carlo |e verfasserin |4 aut | |
| 700 | 1 | |a Iaccino, Enrico |e verfasserin |4 aut | |
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