The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood–brain barrier
As researchers across the globe have focused their attention on understanding SARS-CoV-2, the picture that is emerging is that of a virus that has serious effects on the vasculature in multiple organ systems including the cerebral vasculature. Observed effects on the central nervous system include n...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2020-12-01
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| Series: | Neurobiology of Disease |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S096999612030406X |
| _version_ | 1818651611683094528 |
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| author | Tetyana P. Buzhdygan Brandon J. DeOre Abigail Baldwin-Leclair Trent A. Bullock Hannah M. McGary Jana A. Khan Roshanak Razmpour Jonathan F. Hale Peter A. Galie Raghava Potula Allison M. Andrews Servio H. Ramirez |
| author_facet | Tetyana P. Buzhdygan Brandon J. DeOre Abigail Baldwin-Leclair Trent A. Bullock Hannah M. McGary Jana A. Khan Roshanak Razmpour Jonathan F. Hale Peter A. Galie Raghava Potula Allison M. Andrews Servio H. Ramirez |
| author_sort | Tetyana P. Buzhdygan |
| collection | DOAJ |
| description | As researchers across the globe have focused their attention on understanding SARS-CoV-2, the picture that is emerging is that of a virus that has serious effects on the vasculature in multiple organ systems including the cerebral vasculature. Observed effects on the central nervous system include neurological symptoms (headache, nausea, dizziness), fatal microclot formation and in rare cases encephalitis. However, our understanding of how the virus causes these mild to severe neurological symptoms and how the cerebral vasculature is impacted remains unclear. Thus, the results presented in this report explored whether deleterious outcomes from the SARS-CoV-2 viral spike protein on primary human brain microvascular endothelial cells (hBMVECs) could be observed. The spike protein, which plays a key role in receptor recognition, is formed by the S1 subunit containing a receptor binding domain (RBD) and the S2 subunit. First, using postmortem brain tissue, we show that the angiotensin converting enzyme 2 or ACE2 (a known binding target for the SARS-CoV-2 spike protein), is ubiquitously expressed throughout various vessel calibers in the frontal cortex. Moreover, ACE2 expression was upregulated in cases of hypertension and dementia. ACE2 was also detectable in primary hBMVECs maintained under cell culture conditions. Analysis of cell viability revealed that neither the S1, S2 or a truncated form of the S1 containing only the RBD had minimal effects on hBMVEC viability within a 48 h exposure window. Introduction of spike proteins to in vitro models of the blood-brain barrier (BBB) showed significant changes to barrier properties. Key to our findings is the demonstration that S1 promotes loss of barrier integrity in an advanced 3D microfluidic model of the human BBB, a platform that more closely resembles the physiological conditions at this CNS interface. Evidence provided suggests that the SARS-CoV-2 spike proteins trigger a pro-inflammatory response on brain endothelial cells that may contribute to an altered state of BBB function. Together, these results are the first to show the direct impact that the SARS-CoV-2 spike protein could have on brain endothelial cells; thereby offering a plausible explanation for the neurological consequences seen in COVID-19 patients. |
| first_indexed | 2024-12-17T02:08:52Z |
| format | Article |
| id | doaj.art-8b09ad12a1a144d6870c6f939be164bc |
| institution | Directory Open Access Journal |
| issn | 1095-953X |
| language | English |
| last_indexed | 2024-12-17T02:08:52Z |
| publishDate | 2020-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neurobiology of Disease |
| spelling | doaj.art-8b09ad12a1a144d6870c6f939be164bc2022-12-21T22:07:38ZengElsevierNeurobiology of Disease1095-953X2020-12-01146105131The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood–brain barrierTetyana P. Buzhdygan0Brandon J. DeOre1Abigail Baldwin-Leclair2Trent A. Bullock3Hannah M. McGary4Jana A. Khan5Roshanak Razmpour6Jonathan F. Hale7Peter A. Galie8Raghava Potula9Allison M. Andrews10Servio H. Ramirez11Department of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of America; Center for Substance Abuse Research, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of AmericaDepartment of Biomedical Engineering, Rowan University, Glassboro, NJ 08028, United States of AmericaDepartment of Biomedical Engineering, Rowan University, Glassboro, NJ 08028, United States of AmericaDepartment of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of America; Center for Substance Abuse Research, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of AmericaDepartment of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of AmericaDepartment of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of AmericaDepartment of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of AmericaDepartment of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of AmericaDepartment of Biomedical Engineering, Rowan University, Glassboro, NJ 08028, United States of AmericaDepartment of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of America; Center for Substance Abuse Research, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of AmericaDepartment of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of America; Center for Substance Abuse Research, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of AmericaDepartment of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of America; Center for Substance Abuse Research, The Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, United States of America; The Shriners Hospitals Pediatric Research Center, Philadelphia, PA 19140, United States of America; Corresponding author at: Department of Pathology and Laboratory Medicine, Temple University Lewis Katz School of Medicine, 3500 N, Broad St. MERB 844, Philadelphia, PA 19140, United States of America.As researchers across the globe have focused their attention on understanding SARS-CoV-2, the picture that is emerging is that of a virus that has serious effects on the vasculature in multiple organ systems including the cerebral vasculature. Observed effects on the central nervous system include neurological symptoms (headache, nausea, dizziness), fatal microclot formation and in rare cases encephalitis. However, our understanding of how the virus causes these mild to severe neurological symptoms and how the cerebral vasculature is impacted remains unclear. Thus, the results presented in this report explored whether deleterious outcomes from the SARS-CoV-2 viral spike protein on primary human brain microvascular endothelial cells (hBMVECs) could be observed. The spike protein, which plays a key role in receptor recognition, is formed by the S1 subunit containing a receptor binding domain (RBD) and the S2 subunit. First, using postmortem brain tissue, we show that the angiotensin converting enzyme 2 or ACE2 (a known binding target for the SARS-CoV-2 spike protein), is ubiquitously expressed throughout various vessel calibers in the frontal cortex. Moreover, ACE2 expression was upregulated in cases of hypertension and dementia. ACE2 was also detectable in primary hBMVECs maintained under cell culture conditions. Analysis of cell viability revealed that neither the S1, S2 or a truncated form of the S1 containing only the RBD had minimal effects on hBMVEC viability within a 48 h exposure window. Introduction of spike proteins to in vitro models of the blood-brain barrier (BBB) showed significant changes to barrier properties. Key to our findings is the demonstration that S1 promotes loss of barrier integrity in an advanced 3D microfluidic model of the human BBB, a platform that more closely resembles the physiological conditions at this CNS interface. Evidence provided suggests that the SARS-CoV-2 spike proteins trigger a pro-inflammatory response on brain endothelial cells that may contribute to an altered state of BBB function. Together, these results are the first to show the direct impact that the SARS-CoV-2 spike protein could have on brain endothelial cells; thereby offering a plausible explanation for the neurological consequences seen in COVID-19 patients.http://www.sciencedirect.com/science/article/pii/S096999612030406XCOVID-19SARS-CoV-2Cerebral vascular biologyBlood-brain barrierNeuroinflammationMicrofluidic chip |
| spellingShingle | Tetyana P. Buzhdygan Brandon J. DeOre Abigail Baldwin-Leclair Trent A. Bullock Hannah M. McGary Jana A. Khan Roshanak Razmpour Jonathan F. Hale Peter A. Galie Raghava Potula Allison M. Andrews Servio H. Ramirez The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood–brain barrier Neurobiology of Disease COVID-19 SARS-CoV-2 Cerebral vascular biology Blood-brain barrier Neuroinflammation Microfluidic chip |
| title | The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood–brain barrier |
| title_full | The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood–brain barrier |
| title_fullStr | The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood–brain barrier |
| title_full_unstemmed | The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood–brain barrier |
| title_short | The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood–brain barrier |
| title_sort | sars cov 2 spike protein alters barrier function in 2d static and 3d microfluidic in vitro models of the human blood brain barrier |
| topic | COVID-19 SARS-CoV-2 Cerebral vascular biology Blood-brain barrier Neuroinflammation Microfluidic chip |
| url | http://www.sciencedirect.com/science/article/pii/S096999612030406X |
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