Comparison of SARS-CoV-2 Receptors Expression in Primary Endothelial Cells and Retinoic Acid-Differentiated Human Neuronal Cells
Francesca Benedetti,
Giovannino Silvestri,
Carla Mavian,
Matthew Weichseldorfer,
Arshi Munawwar,
Melanie N. Cash,
Melissa Dulcey,
Amy Y. Vittor,
Massimo Ciccozzi,
Marco Salemi,
Olga S. Latinovic,
Davide Zella
Affiliations
Francesca Benedetti
Institute of Human Virology and Global Virus Network Center, Department of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USA
Giovannino Silvestri
Institute of Human Virology and Global Virus Network Center, Department of Medicine, School of Medicine, University of Maryland, Baltimore, MD 21201, USA
Carla Mavian
Emerging Pathogens Institute, Department of Pathology, Immunology and Laboratory Medicine, College of Medicine, University of Florida, Gainesville, FL 32610, USA
Matthew Weichseldorfer
Institute of Human Virology and Global Virus Network Center, Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, MD 21201, USA
Arshi Munawwar
Institute of Human Virology and Global Virus Network Center, Department of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USA
Melanie N. Cash
Emerging Pathogens Institute, Department of Pathology, Immunology and Laboratory Medicine, College of Medicine, University of Florida, Gainesville, FL 32610, USA
Melissa Dulcey
Emerging Pathogens Institute, Department of Environmental and Global Health, College of Medicine, University of Florida, Gainesville, FL 32610, USA
Amy Y. Vittor
Emerging Pathogens Institute, Department of Environmental and Global Health, College of Medicine, University of Florida, Gainesville, FL 32610, USA
Massimo Ciccozzi
Medical Statistic and Molecular Epidemiology Unit, University of Biomedical Campus, 00128 Rome, Italy
Marco Salemi
Emerging Pathogens Institute, Department of Pathology, Immunology and Laboratory Medicine, College of Medicine, University of Florida, Gainesville, FL 32610, USA
Olga S. Latinovic
Institute of Human Virology and Global Virus Network Center, Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, MD 21201, USA
Davide Zella
Institute of Human Virology and Global Virus Network Center, Department of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USA
SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) is primarily responsible for coronavirus disease (COVID-19) and it is characterized by respiratory illness with fever and dyspnea. Severe vascular problems and several other manifestations, including neurological ones, have also been frequently reported, particularly in the great majority of “long hauler” patients. SARS-CoV-2 infects and replicates in lung epithelial cells, while dysfunction of endothelial and neuronal brain cells has been observed in the absence of productive infection. It has been shown that the Spike protein can interact with specific cellular receptors, supporting both viral entry and cellular dysfunction. It is thus clear that understanding how and when these receptors are regulated, as well as how much they are expressed would help in unveiling the multifaceted aspects of this disease. Here, we show that SH-SY5Y neuroblastoma cells express three important cellular surface molecules that interact with the Spike protein, namely ACE2, TMPRSS2, and NRP1. Their levels increase when cells are treated with retinoic acid (RA), a commonly used agent known to promote differentiation. This increase matched the higher levels of receptors observed on HUVEC (primary human umbilical vein endothelial cells). We also show by confocal imaging that replication-defective pseudoviruses carrying the SARS-CoV-2 Spike protein can infect differentiated and undifferentiated SH-SY5Y, and HUVEC cells, although with different efficiencies. Neuronal cells and endothelial cells are potential targets for SARS-CoV-2 infection and the interaction of the Spike viral protein with these cells may cause their dysregulation. Characterizing RNA and protein expression tempo, mode, and levels of different SARS-CoV-2 receptors on both cell subpopulations may have clinical relevance for the diagnosis and treatment of COVID-19-infected subjects, including long hauler patients with neurological manifestations.
