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      Circulating microRNA signatures associated with disease severity and outcome in COVID-19 patients

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          Abstract

          Background

          SARS-CoV-2 induces a spectrum of clinical conditions ranging from asymptomatic infection to life threatening severe disease. Host microRNAs have been involved in the cytokine storm driven by SARS-CoV-2 infection and proposed as candidate biomarkers for COVID-19.

          Methods

          To discover signatures of circulating miRNAs associated with COVID-19, disease severity and mortality, small RNA-sequencing was performed on serum samples collected from 89 COVID-19 patients (34 severe, 29 moderate, 26 mild) at hospital admission and from 45 healthy controls (HC). To search for possible sources of miRNAs, investigation of differentially expressed (DE) miRNAs in relevant human cell types in vitro.

          Results

          COVID-19 patients showed upregulation of miRNAs associated with lung disease, vascular damage and inflammation and downregulation of miRNAs that inhibit pro-inflammatory cytokines and chemokines, angiogenesis, and stress response. Compared with mild/moderate disease, patients with severe COVID-19 had a miRNA signature indicating a profound impairment of innate and adaptive immune responses, inflammation, lung fibrosis and heart failure. A subset of the DE miRNAs predicted mortality. In particular, a combination of high serum miR-22-3p and miR-21-5p, which target antiviral response genes, and low miR-224-5p and miR-155-5p, targeting pro-inflammatory factors, discriminated severe from mild/moderate COVID-19 (AUROC 0.88, 95% CI 0.80-0.95, p<0.0001), while high leukocyte count and low levels of miR-1-3p, miR-23b-3p, miR-141-3p, miR-155-5p and miR-4433b-5p predicted mortality with high sensitivity and specificity (AUROC 0.95, 95% CI 0.89-1.00, p<0.0001). In vitro experiments showed that some of the DE miRNAs were modulated directly by SARS-CoV-2 infection in permissive lung epithelial cells.

          Conclusions

          We discovered circulating miRNAs associated with COVID-19 severity and mortality. The identified DE miRNAs provided clues on COVID-19 pathogenesis, highlighting signatures of impaired interferon and antiviral responses, inflammation, organ damage and cardiovascular failure as associated with severe disease and death.

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          Most cited references78

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          Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2

          In comparative high-throughput sequencing assays, a fundamental task is the analysis of count data, such as read counts per gene in RNA-seq, for evidence of systematic changes across experimental conditions. Small replicate numbers, discreteness, large dynamic range and the presence of outliers require a suitable statistical approach. We present DESeq2, a method for differential analysis of count data, using shrinkage estimation for dispersions and fold changes to improve stability and interpretability of estimates. This enables a more quantitative analysis focused on the strength rather than the mere presence of differential expression. The DESeq2 package is available at http://www.bioconductor.org/packages/release/bioc/html/DESeq2.html. Electronic supplementary material The online version of this article (doi:10.1186/s13059-014-0550-8) contains supplementary material, which is available to authorized users.
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            Cytoscape: a software environment for integrated models of biomolecular interaction networks.

            Cytoscape is an open source software project for integrating biomolecular interaction networks with high-throughput expression data and other molecular states into a unified conceptual framework. Although applicable to any system of molecular components and interactions, Cytoscape is most powerful when used in conjunction with large databases of protein-protein, protein-DNA, and genetic interactions that are increasingly available for humans and model organisms. Cytoscape's software Core provides basic functionality to layout and query the network; to visually integrate the network with expression profiles, phenotypes, and other molecular states; and to link the network to databases of functional annotations. The Core is extensible through a straightforward plug-in architecture, allowing rapid development of additional computational analyses and features. Several case studies of Cytoscape plug-ins are surveyed, including a search for interaction pathways correlating with changes in gene expression, a study of protein complexes involved in cellular recovery to DNA damage, inference of a combined physical/functional interaction network for Halobacterium, and an interface to detailed stochastic/kinetic gene regulatory models.
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              Cutadapt removes adapter sequences from high-throughput sequencing reads

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                Author and article information

                Contributors
                Journal
                Front Immunol
                Front Immunol
                Front. Immunol.
                Frontiers in Immunology
                Frontiers Media S.A.
                1664-3224
                11 August 2022
                2022
                11 August 2022
                : 13
                : 968991
                Affiliations
                [1] 1 Department of Medicine, University of Padova , Padova, Italy
                [2] 2 Department of Molecular Medicine, University of Padova , Padova, Italy
                [3] 3 Department of Infectious-Tropical Diseases and Microbiology, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Sacro Cuore Don Calabria Hospital , Verona, Italy
                [4] 4 Maternal and Child Health Department, University of Padova , Padova, Italy
                [5] 5 Microbiology and Virology Unit, Padova University Hospital , Padova, Italy
                Author notes

                Edited by: Nitin Saksena, Victoria University, Australia

                Reviewed by: Vitale Miceli, Mediterranean Institute for Transplantation and Highly Specialized Therapies (ISMETT), Italy; Samuel Brennan, Genieus Genomics, Australia

                *Correspondence: Luisa Barzon, luisa.barzon@ 123456unipd.it

                †These authors have contributed equally to this work and share first authorship

                ‡These authors share last authorship

                This article was submitted to Vaccines and Molecular Therapeutics, a section of the journal Frontiers in Immunology

                Article
                10.3389/fimmu.2022.968991
                9403711
                36032130
                92ba172b-84b5-421d-8128-f9ae0face83d
                Copyright © 2022 Giannella, Riccetti, Sinigaglia, Piubelli, Razzaboni, Di Battista, Agostini, Dal Molin, Manganelli, Gobbi, Ceolotto and Barzon

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 14 June 2022
                : 20 July 2022
                Page count
                Figures: 8, Tables: 3, Equations: 0, References: 79, Pages: 21, Words: 10980
                Funding
                Funded by: Horizon 2020 , doi 10.13039/501100007601;
                Award ID: 874735
                Funded by: Università degli Studi di Padova , doi 10.13039/501100003500;
                Award ID: PRID-2020
                Funded by: Ministero della Salute , doi 10.13039/501100003196;
                Award ID: Fondi Ricerca Corrente – L1P6, COVID-2020-12371675
                Categories
                Immunology
                Original Research

                Immunology
                covid-19,microrna,biomarkers,innate immunity,inflammation,interferon,sars-cov-2,rna-sequencing
                Immunology
                covid-19, microrna, biomarkers, innate immunity, inflammation, interferon, sars-cov-2, rna-sequencing

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