tetano
Editor, Senior Moderator
Stroke
. 2020 Nov 9;STROKEAHA120032764.
doi: 10.1161/STROKEAHA.120.032764. Online ahead of print.
Flow-Mediated Susceptibility and Molecular Response of Cerebral Endothelia to SARS-CoV-2 Infection
Naoki Kaneko[SUP] #[/SUP][SUP] 1 [/SUP], Sandro Satta[SUP] #[/SUP][SUP] 2 [/SUP], Yutaro Komuro[SUP] 3 [/SUP], Sree Deepthi Muthukrishnan[SUP] 4 [/SUP], Visesha Kakarla[SUP] 5 [/SUP], Lea Guo[SUP] 1 [/SUP], Jennifer An[SUP] 3 [/SUP], Fanny Elahi[SUP] 6 [/SUP], Harley I Kornblum[SUP] 4 [/SUP], David S Liebeskind[SUP] 3 [/SUP], Tzung Hsiai[SUP] 2 7 [/SUP], Jason D Hinman[SUP] 3 8 [/SUP]
Affiliations
Abstract
Background and purpose: Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection is associated with an increased rate of cerebrovascular events including ischemic stroke and intracerebral hemorrhage. The mechanisms underlying cerebral endothelial susceptibility and response to SARS-CoV-2 are unknown yet critical to understanding the association of SARS-CoV-2 infection with cerebrovascular events.
Methods: Endothelial cells were isolated from human brain and analyzed by RNA sequencing. Human umbilical vein and human brain microvascular cells were used in both monolayer culture and endothelialized within a 3-dimensional printed vascular model of the middle cerebral artery. Gene expression levels were measured by quantitative polymerase chain reaction and direct RNA hybridization. Recombinant SARS-CoV-2 S protein and S protein-containing liposomes were used to measure endothelial binding by immunocytochemistry.
Results: ACE2 (angiotensin-converting enzyme-2) mRNA levels were low in human brain and monolayer endothelial cell culture. Within the 3-dimensional printed vascular model, ACE2 gene expression and protein levels were progressively increased by vessel size and flow rates. SARS-CoV-2 S protein-containing liposomes were detected in human umbilical vein endothelial cells and human brain microvascular endothelial cells in 3-dimensional middle cerebral artery models but not in monolayer culture consistent with flow dependency of ACE2 expression. Binding of SARS-CoV-2 S protein triggered 83 unique genes in human brain endothelial cells including upregulation of complement component C3.
Conclusions: Brain endothelial cells are susceptible to direct SARS-CoV-2 infection through flow-dependent expression of ACE2. Viral S protein binding triggers a unique gene expression profile in brain endothelia that may explain the association of SARS-CoV-2 infection with cerebrovascular events.
Keywords: cerebrovascular circulation; endothelial cells; endothelium, vascular; models, theoretical; viruses.
. 2020 Nov 9;STROKEAHA120032764.
doi: 10.1161/STROKEAHA.120.032764. Online ahead of print.
Flow-Mediated Susceptibility and Molecular Response of Cerebral Endothelia to SARS-CoV-2 Infection
Naoki Kaneko[SUP] #[/SUP][SUP] 1 [/SUP], Sandro Satta[SUP] #[/SUP][SUP] 2 [/SUP], Yutaro Komuro[SUP] 3 [/SUP], Sree Deepthi Muthukrishnan[SUP] 4 [/SUP], Visesha Kakarla[SUP] 5 [/SUP], Lea Guo[SUP] 1 [/SUP], Jennifer An[SUP] 3 [/SUP], Fanny Elahi[SUP] 6 [/SUP], Harley I Kornblum[SUP] 4 [/SUP], David S Liebeskind[SUP] 3 [/SUP], Tzung Hsiai[SUP] 2 7 [/SUP], Jason D Hinman[SUP] 3 8 [/SUP]
Affiliations
- PMID: 33161843
- DOI: 10.1161/STROKEAHA.120.032764
Abstract
Background and purpose: Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection is associated with an increased rate of cerebrovascular events including ischemic stroke and intracerebral hemorrhage. The mechanisms underlying cerebral endothelial susceptibility and response to SARS-CoV-2 are unknown yet critical to understanding the association of SARS-CoV-2 infection with cerebrovascular events.
Methods: Endothelial cells were isolated from human brain and analyzed by RNA sequencing. Human umbilical vein and human brain microvascular cells were used in both monolayer culture and endothelialized within a 3-dimensional printed vascular model of the middle cerebral artery. Gene expression levels were measured by quantitative polymerase chain reaction and direct RNA hybridization. Recombinant SARS-CoV-2 S protein and S protein-containing liposomes were used to measure endothelial binding by immunocytochemistry.
Results: ACE2 (angiotensin-converting enzyme-2) mRNA levels were low in human brain and monolayer endothelial cell culture. Within the 3-dimensional printed vascular model, ACE2 gene expression and protein levels were progressively increased by vessel size and flow rates. SARS-CoV-2 S protein-containing liposomes were detected in human umbilical vein endothelial cells and human brain microvascular endothelial cells in 3-dimensional middle cerebral artery models but not in monolayer culture consistent with flow dependency of ACE2 expression. Binding of SARS-CoV-2 S protein triggered 83 unique genes in human brain endothelial cells including upregulation of complement component C3.
Conclusions: Brain endothelial cells are susceptible to direct SARS-CoV-2 infection through flow-dependent expression of ACE2. Viral S protein binding triggers a unique gene expression profile in brain endothelia that may explain the association of SARS-CoV-2 infection with cerebrovascular events.
Keywords: cerebrovascular circulation; endothelial cells; endothelium, vascular; models, theoretical; viruses.