TY - JOUR
T1 - ACE-2, TMPRSS2, and Neuropilin-1 Receptor Expression on Human Brain Astrocytes and Pericytes and SARS-CoV-2 Infection Kinetics
AU - Malik, Johid Reza
AU - Acharya, Arpan
AU - Avedissian, Sean N.
AU - Byrareddy, Siddappa N.
AU - Fletcher, Courtney V.
AU - Podany, Anthony T.
AU - Dyavar, Shetty Ravi
N1 - Funding Information:
This work was supported by UNMC Nebraska Neuroscience Alliance COVID-19 Rapid Response grant to S.R.D, C.V.F, A.T.P, and S.N.A; NIMH P30 MH062261 and Nebraska Research Initiative (NRI) Collaborative grant awarded to S.R.D, A.T.P and C.V.F; K23AI134307-01A1 (to A.T.P); R01-AI124965 (to C.V.F.) by the National Institute of Allergy and Infectious Diseases (NIAID) of the National Institutes of Health (NIH), Bethesda, MD, USA and K23 MH125734 (to SNA). S.N.B acknowledges independent research and development (IRAD) funding from the National Strategic Research Institute (NSRI), Nebraska Research Initiative (NRI) grants at the University of Nebraska. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Funding Information:
We acknowledge the UNMC ABSL3, and BSL-3 core facility for allowing us to perform all in vivo and invitro experiments involving SARS-CoV-2. This facility is administered by the office of the Vice-Chancellor for research and supported by the Nebraska Research Initiative (NRI). We acknowledge all the current and past members of S.N.B’s laboratory for helping to complete the experiments described in the manuscript. The following reagents were deposited by the Centers for Disease Control and Prevention and obtained through BEI Resources, NIAID, NIH: (a) SARS-Related Coronavirus 2, Isolate USA-WI1/2020, and (b) Quantitative PCR (qPCR) Control RNA from Heat-Inactivated SARS-Related Coronavirus 2, Isolate USA-WA1/2020, NR 52347. All individuals included in this section have consented to the acknowledgement.
Publisher Copyright:
© 2023 by the authors.
PY - 2023/5
Y1 - 2023/5
N2 - Angiotensin Converting Enzyme 2 (ACE-2), Transmembrane Serine Protease 2 (TMPRSS-2) and Neuropilin-1 cellular receptors support the entry of SARS-CoV-2 into susceptible human target cells and are characterized at the molecular level. Some evidence on the expression of entry receptors at mRNA and protein levels in brain cells is available, but co-expression of these receptors and confirmatory evidence on brain cells is lacking. SARS-CoV-2 infects some brain cell types, but infection susceptibility, multiple entry receptor density, and infection kinetics are rarely reported in specific brain cell types. Highly sensitive Taqman ddPCR, flow-cytometry and immunocytochemistry assays were used to quantitate the expression of ACE-2, TMPRSS-2 and Neuropilin-1 at mRNA and protein levels on human brain-extracted pericytes and astrocytes, which are an integral part of the Blood-Brain-Barrier (BBB). Astrocytes showed moderate ACE-2 (15.9 ± 1.3%, Mean ± SD, n = 2) and TMPRSS-2 (17.6%) positive cells, and in contrast show high Neuropilin-1 (56.4 ± 39.8%, n = 4) protein expression. Whereas pericytes showed variable ACE-2 (23.1 ± 20.7%, n = 2), Neuropilin-1 (30.3 ± 7.5%, n = 4) protein expression and higher TMPRSS-2 mRNA (667.2 ± 232.3, n = 3) expression. Co-expression of multiple entry receptors on astrocytes and pericytes allows entry of SARS-CoV-2 and progression of infection. Astrocytes showed roughly four-fold more virus in culture supernatants than pericytes. SARS-CoV-2 cellular entry receptor expression and “in vitro” viral kinetics in astrocytes and pericytes may improve our understanding of viral infection “in vivo”. In addition, this study may facilitate the development of novel strategies to counter the effects of SARS-CoV-2 and inhibit viral infection in brain tissues to prevent the spread and interference in neuronal functions.
AB - Angiotensin Converting Enzyme 2 (ACE-2), Transmembrane Serine Protease 2 (TMPRSS-2) and Neuropilin-1 cellular receptors support the entry of SARS-CoV-2 into susceptible human target cells and are characterized at the molecular level. Some evidence on the expression of entry receptors at mRNA and protein levels in brain cells is available, but co-expression of these receptors and confirmatory evidence on brain cells is lacking. SARS-CoV-2 infects some brain cell types, but infection susceptibility, multiple entry receptor density, and infection kinetics are rarely reported in specific brain cell types. Highly sensitive Taqman ddPCR, flow-cytometry and immunocytochemistry assays were used to quantitate the expression of ACE-2, TMPRSS-2 and Neuropilin-1 at mRNA and protein levels on human brain-extracted pericytes and astrocytes, which are an integral part of the Blood-Brain-Barrier (BBB). Astrocytes showed moderate ACE-2 (15.9 ± 1.3%, Mean ± SD, n = 2) and TMPRSS-2 (17.6%) positive cells, and in contrast show high Neuropilin-1 (56.4 ± 39.8%, n = 4) protein expression. Whereas pericytes showed variable ACE-2 (23.1 ± 20.7%, n = 2), Neuropilin-1 (30.3 ± 7.5%, n = 4) protein expression and higher TMPRSS-2 mRNA (667.2 ± 232.3, n = 3) expression. Co-expression of multiple entry receptors on astrocytes and pericytes allows entry of SARS-CoV-2 and progression of infection. Astrocytes showed roughly four-fold more virus in culture supernatants than pericytes. SARS-CoV-2 cellular entry receptor expression and “in vitro” viral kinetics in astrocytes and pericytes may improve our understanding of viral infection “in vivo”. In addition, this study may facilitate the development of novel strategies to counter the effects of SARS-CoV-2 and inhibit viral infection in brain tissues to prevent the spread and interference in neuronal functions.
KW - ACE-2
KW - COVID-19
KW - Neuropilin-1
KW - SARS-CoV-2
KW - TMPRSS-2
KW - astrocytes
KW - brain microvascular endothelial cells and blood-brain-barrier
KW - pericytes
UR - http://www.scopus.com/inward/record.url?scp=85160379715&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85160379715&partnerID=8YFLogxK
U2 - 10.3390/ijms24108622
DO - 10.3390/ijms24108622
M3 - Article
C2 - 37239978
AN - SCOPUS:85160379715
SN - 1661-6596
VL - 24
JO - International journal of molecular sciences
JF - International journal of molecular sciences
IS - 10
M1 - 8622
ER -