SARS-CoV-2 Infection Triggers Auto-Immune Response in ARDS

Acute respiratory distress syndrome (ARDS) is a severe pulmonary disease, which is one of the major complications in COVID-19 patients. Dysregulation of the immune system and imbalances in cytokine release and immune cell activation are involved in SARS-CoV-2 infection. Here, the inflammatory, antigen, and auto-immune profile of patients presenting COVID-19-associated severe ARDS has been analyzed using functional proteomics approaches. Both, innate and humoral responses have been characterized through acute-phase protein network and auto-antibody signature. Severity and sepsis by SARS-CoV-2 emerged to be correlated with auto-immune profiles of patients and define their clinical progression, which could provide novel perspectives in therapeutics development and biomarkers of COVID-19 patients. Humoral response in COVID-19 patients’ profile separates with significant differences patients with or without ARDS. Furthermore, we found that this profile can be correlated with COVID-19 severity and results more common in elderly patients.We gratefully acknowledge financial support from the Spanish Health Institute Carlos III (ISCIII) for the grants: FIS PI14/01538, FIS PI17/01930, and CB16/12/00400. This research work was also funded by the European Commission – NextGenerationEU, through CSIC's Global Health Platform (PTI Salud Global) We also acknowledge Fondos FEDER (EU) and Junta Castilla-León (COVID19 grant COV20EDU/00187). The Proteomics Unit belongs to ProteoRed, PRB3-ISCIII, supported by grant PT17/0019/0023, of the PE I + D + I 2017-2020, funded by ISCIII and FEDER. AL-V is supported by the VIII Centenario-USAL PhD Program. PJ-V is supported by the JCYL PhD Program “Nos Impulsa-JCYL” and scholarship JCYL-EDU/601/2020. EXOHEP-CM S2017/BMD3727 by Comunidad de Madrid and Fondos FEDER (to JL and RH) and PI19/01091 by ISCIII (to RH).Peer reviewe

Transcriptional profiling and genotyping of degraded nucleic acids from autopsy tissue samples after prolonged formalin fixation times

[Background]: Samples used for genotyping and transcription studies are obtained and conserved in very specific conditions. The possibility to use autopsy tissue samples, which contain nucleic acids of very poor quality, would open new possibilities for genetic studies. [Methods]: We have used liver tissue samples from autopsy cases to (i) determine its quality; (ii) study gene expression of 13 genes involved in different cell processes, before and after cDNA pre-amplification (quantitative reverse transcriptase polymerase chain reaction); and (iii) analyze the presence of 2 common polymorphisms of relevance for illness (ACE I/D genotype by PCR amplification, and TNF-α promoter gene polymorphism, by DNA sequencing). [Results]: Samples were grouped according to different buffered formalin fixation times (group 1, <15 days; group 2, 60-90 days; group 3, 150-180 days; group 4, 240-270 days). Nucleic acids showed a time-dependent degradation. The expression of 13 genes could be studied in all cases from groups 1 and 2, only 7 from group 3 and none from group 4. cDNA preamplification allowed the study of all genes in all samples. DNA genotyping for ACE and TNF-α promoter region was possible in all cases. [Conclusions]: We conclude that nucleic acids extracted from autopsy specimens after prolonged periods of time in formalin were of sufficient quality to study gene expression and genotyping using currently available methodology and cDNA pre-amplification.This study was funded by FIS PI04/08/42, Instituto de Salud Carlos III, Spain.Peer Reviewe

MicroRNAs as biomarkers of acute lung injury

Acute respiratory distress syndrome (ARDS) is a common and complex inflammatory lung diseases affecting critically ill patients requiring mechanical ventilation. MicroRNAs (miRNAs), a novel pathway of non-coding RNA molecules that regulate gene expression at the post-transcriptional level, have emerged as a novel class of gene expression, and can play important roles in inflammation or apoptosis, which are common manifestations of ARDS and diffuse alveolar damage (DAD). In the present review, we discuss the role of miRNAs as biomarkers of ARDS and DAD, and their potential use as therapeutic targets for this condition.Sin financiación3.689 JCR (2018) Q1, 121/244 Oncology0.958 SJR (2018) Q1, 602/2844 Medicine (miscellaneous)No data IDR 2018UE

Inhibition of cholesterol biosynthesis by Delta22-unsaturated phytosterols via competitive inhibition of sterol Delta24-reductase in mammalian cells.

Dietary phytosterols are cholesterol-lowering agents that interfere with the intestinal absorption of cholesterol. In the present study, we have studied their effects on cholesterol biosynthesis in human cells, particularly in the sterol-conversion pathway. For this, both Caco-2 (intestinal mucosa) and HL-60 (promyelocytic) human cell lines were incubated with [(14)C]acetate, and the incorporation of radioactivity into sterols was determined using HPLC and radioactivity detection online. Sterols containing a double bond at C-22 in the side chain (stigmasterol, brassicasterol and ergosterol) dramatically inhibited the activity of sterol Delta(24)-reductase, as indicated by the decrease in radioactivity incorporation into cholesterol and the accumulation of its precursors (mainly desmosterol). Phytosterols with the saturated side chain (beta-sitosterol and campesterol) were inactive in this regard. The inhibition of sterol (24)-reductase was confirmed in rat liver microsomes by using (14)C-labelled desmosterol as the substrate. The (22)-unsaturated phytosterols acted as competitive inhibitors of sterol (24)-reductase, with K(i) values (41.1, 42.7 and 36.8 microM for stigmasterol, brassicasterol and ergosterol respectively) similar to the estimated K(m) for desmosterol (26.3 microM). The sterol 5,22-cholestedien-3beta-ol, an unusual desmosterol isomer that lacks the alkyl groups characteristic of phytosterols, acted as a much stronger inhibitor of (24)-reductase (K(i)=3.34 microM). The usually low intracellular concentrations of the physiological substrates of (24)-reductase explains the strong inhibition of cholesterol biosynthesis that these compounds exert in cells. Given that inhibition of sterol (24)-reductase was achieved at physiologically relevant concentrations, it may represent an additional mechanism for the cholesterol-lowering action of phytosterols, and opens up the possibility of using certain (22)-unsaturated sterols as effective hypocholesterolaemic agents

Characteristics of microRNAs and their potential relevance for the diagnosis and therapy of the acute respiratory distress syndrome: from bench to bedside

Acute respiratory distress syndrome (ARDS) is a complex disease associated with high morbidity and mortality. Biomarkers and specific pharmacologic treatment of the syndrome are lacking. MicroRNAs (miRNAs) are small (∼19–22 nucleotides) noncoding RNA molecules whose function is the regulation of gene expression. Their uncommon biochemical characteristics (eg, their resistance to degradation because of extreme temperature and pH fluctuations, freeze-thaw cycles, long storage times in frozen conditions, and RNAse digestion) and their presence in a wide range of different biological fluids and the relatively low number of individual miRNAs make these molecules good biomarkers in different clinical conditions. In addition, miRNAs are suitable therapeutic targets as their expression can be modulated by different available strategies. The aim of the present review is to offer clinicians a global perspective of miRNA, covering their structure and nomenclature, biogenesis, effects on gene expression, regulation of expression, and features as disease biomarkers and therapeutic targets, with special attention to ARDS. Because of the early stage of research on miRNAs applied to ARDS, attention has been focused on how knowledge sourced from basic and translational research could inspire future clinical studies.Sin financiación4.652 JCR (2016) Q1, 4/30 Medical Laboratory Technology, 17/155 Medicine, General and Internal, 19/128 Medicine, Research and ExperimentalUE

Microrna 155-3P modulates the FASL-mediated inflammation in human alveolar epithelial cells in vitro

PI15-00482, PI15-1942.2.615 JCR (2019) Q2, 10/29 Medical Laboratory Technology0.876 SJR (2019) Q1, 661/2806 Medicine (miscellaneous)No data IDR 2019UE

Does the Inflammatory Response Differ in Septic and Non-Septic Insults?

Sin financiación15.239 JCR (2017) Q1, 2/60 Respiratory System, 2/33 Critical Care MedicineUE

Sepsis and mechanical ventilation alter the tight junction proteins in rat lungs in vivo

Introduction: The tight junction (TJ) complexes of the alveolar epithelium regulate lung permeability. Objectives: To determine whether sepsis and mechanical ventilation alter the TJ complexes in the alveolar walls of rat lungs in vivo. Methods: We performed an abdominal sepsis model induced by cecal ligation and puncture (CLP) followed 24 h later by mechanical ventilation (MV) with a protective strategy (Vt= 9 ml/kg+PEEP 5 cmH2O for 4 h) in adult rats (Sprague-Dawley). The groups were (n= 10/group): Sham-without MV (laparotomy only, control group), CLP-without MV, Sham+MV and CLP+MV. We collected lung tissue, bronchoalveolar lavage fluid (BALF) and plasma and we measured the levels of Occludin and ZO1 (ELISA, inmunofluorescence), cytokines (ELISA) and apoptosis (caspase 3 and TUNEL). Results: 1) The levels of cytokines were similar in all groups, except for a decrease of IL1b in the BALF of the CLP+MV group. 2) Compared with the control group, the activity of caspase 3 and apoptosis in rat lungs was increased in the CLP, MV and CLP+MV groups. 3) Compared with the control group, the levels of occludin were increased in the lungs of rats with CLP (with or without MV). Only the combination of CLP and MV increased the levels of occludin in BALF and plasma. In contrast, ZO1 levels decreased in rat lungs after CLP, MV or CLP+MV. In CLP-without MV rats, ZO1 levels increased in the BALF and decreased in plasma. Conclusions: Abdominal sepsis and MV with a protective strategy cause apoptosis and alter the expression of TJ proteins in the lung without modifying the expression of cytokines. Changes in the levels of the TJ proteins in BALF and plasma could be used as diagnostic biomarkers in acute lung injury.Sin financiación12.339 JCR (2019) Q1, 4/64 Respiratory System3.398 SJR (2019) Q1, 5/149 Pulmonary and Respiratory MedicineNo data IDR 2019UE