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Protein C Pretreatment Protects Endothelial Cells from SARS-CoV-2-Induced Activation
Bruna Rafaela Dos Santos Silva1,2, Davi Sidarta-Oliveira2,3, Joseane Morari2,3
1School of Nursing, Universidade Estadual de Campinas, Campinas 13083-970, SP, Brazil.
Abstract:
SARS-CoV-2 can induce vascular dysfunction and thrombotic events in patients with severe COVID-19; however, the cellular and molecular mechanisms behind these effects remain largely unknown. In this study, we used a combination of experimental and in silico approaches to investigate the role of PC in vascular and thrombotic events in COVID-19. Single-cell RNA-sequencing data from patients with COVID-19 and healthy subjects were obtained from the publicly available Gene Expression Omnibus (GEO) repository. In addition, HUVECs were treated with inactive protein C before exposure to SARS-CoV-2 infection or a severe COVID-19 serum. An RT-qPCR array containing 84 related genes was used, and the candidate genes obtained were evaluated. Activated protein C levels were measured using an ELISA kit. We identified at the single-cell level the expression of several pro-inflammatory and pro-coagulation genes in endothelial cells from the patients with COVID-19. Furthermore, we demonstrated that exposure to SARS-CoV-2 promoted transcriptional changes in HUVECs that were partly reversed by the activated protein C pretreatment. We also observed that the serum of severe COVID-19 had a significant amount of activated protein C that could protect endothelial cells from serum-induced activation. In conclusion, activated protein C protects endothelial cells from pro-inflammatory and pro-coagulant effects during exposure to the SARS-CoV-2 virus.
Insights
Activated protein C protects endothelial cells from SARS-CoV-2 virus effects. This study reveals protein C
Area of Science:
- * Vascular Biology and Virology
- * Endothelial Cell Function
- * Coagulation and Thrombosis
Background:
- * Severe COVID-19 is linked to vascular dysfunction and thrombosis.
- * Mechanisms underlying these effects are not fully understood.
- * The role of protein C in COVID-19-associated vascular events requires investigation.
Purpose of the Study:
- * To investigate the role of protein C in vascular and thrombotic events in COVID-19.
- * To elucidate cellular and molecular mechanisms involved.
- * To assess the protective effects of activated protein C on endothelial cells.
Main Methods:
- * Analysis of single-cell RNA-sequencing data from COVID-19 patients and healthy controls.
- * In vitro experiments using human umbilical vein endothelial cells (HUVECs) treated with protein C and exposed to SARS-CoV-2 or COVID-19 serum.
- * RT-qPCR array for gene expression analysis and ELISA for activated protein C quantification.
Main Results:
- * Identified pro-inflammatory and pro-coagulation gene expression in endothelial cells of COVID-19 patients.
- * SARS-CoV-2 exposure induced transcriptional changes in HUVECs, partly reversed by activated protein C.
- * Severe COVID-19 serum contained significant activated protein C, offering protection to endothelial cells.
Conclusions:
- * Activated protein C demonstrates protective effects on endothelial cells against SARS-CoV-2.
- * Protein C mitigates pro-inflammatory and pro-coagulant responses in endothelial cells during viral exposure.
- * Findings highlight protein C as a potential therapeutic target for COVID-19-related vascular complications.
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