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. 2023 Mar 27;61(8):1518-1524.
doi: 10.1515/cclm-2022-1286. Print 2023 Jul 26.

SARS CoV-2 spike protein-guided exosome isolation facilitates detection of potential miRNA biomarkers in COVID-19 infections

Selena Mimmi  1 , Anna Maria Zimbo  1 , Salvatore Rotundo  2 , Erika Cione  3 , Nancy Nisticò  1 , Annamaria Aloisio  1 , Domenico Maisano  4 , Anna Maria Tolomeo  5 , Vincenzo Dattilo  1 , Rosaria Lionello  2 , Antonella Fioravanti  6 , Antonio Di Loria  7 , Angela Quirino  8 , Nadia Marascio  8 , Alessandro Russo  2 , Enrico Maria Trecarichi  2 , Giovanni Matera  8 , Ileana Quinto  1 , Carlo Torti  2 , Enrico Iaccino  1
Affiliations
Free article

SARS CoV-2 spike protein-guided exosome isolation facilitates detection of potential miRNA biomarkers in COVID-19 infections

Selena Mimmi et al. Clin Chem Lab Med. .
Free article

Abstract

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.

Keywords: ACE2; Covid19; biomarkers; exosomes; miRNAs.

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