Neuropilin-1 facilitates SARS-CoV-2 cell entry and infectivity

Another host factor for SARS-CoV-2 Virus-host interactions determine cellular entry and spreading in tissues. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the earlier SARS-CoV use angiotensin-converting enzyme 2 (ACE2) as a receptor; however, their tissue tropism differs, raising the possibility that additional host factors are involved. The spike protein of SARS-CoV-2 contains a cleavage site for the protease furin that is absent from SARS-CoV (see the Perspective by Kielian). Cantuti-Castelvetri et al. now show that neuropilin-1 (NRP1), which is known to bind furin-cleaved substrates, potentiates SARS-CoV-2 infectivity. NRP1 is abundantly expressed in the respiratory and olfactory epithelium, with highest expression in endothelial and epithelial cells. Daly et al. found that the furin-cleaved S1 fragment of the spike protein binds directly to cell surface NRP1 and blocking this interaction with a small-molecule inhibitor or monoclonal antibodies reduced viral infection in cell culture. Understanding the role of NRP1 in SARS-CoV-2 infection may suggest potential targets for future antiviral therapeutics. Science, this issue p. 856, p. 861; see also p. 765 NRP1 serves as a host factor for SARS-CoV-2 infection and may potentially provide a therapeutic target for COVID-19. The causative agent of coronavirus disease 2019 (COVID-19) is the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). For many viruses, tissue tropism is determined by the availability of virus receptors and entry cofactors on the surface of host cells. In this study, we found that neuropilin-1 (NRP1), known to bind furin-cleaved substrates, significantly potentiates SARS-CoV-2 infectivity, an effect blocked by a monoclonal blocking antibody against NRP1. A SARS-CoV-2 mutant with an altered furin cleavage site did not depend on NRP1 for infectivity. Pathological analysis of olfactory epithelium obtained from human COVID-19 autopsies revealed that SARS-CoV-2 infected NRP1-positive cells facing the nasal cavity. Our data provide insight into SARS-CoV-2 cell infectivity and define a potential target for antiviral intervention.

[1]  P. Lehner,et al.  How does SARS-CoV-2 cause COVID-19? , 2020, Science.

[2]  D. Matthews,et al.  Characterisation of the transcriptome and proteome of SARS-CoV-2 reveals a cell passage induced in-frame deletion of the furin-like cleavage site from the spike glycoprotein , 2020, Genome Medicine.

[3]  David H. Brann,et al.  COVID-19 and the Chemical Senses: Supporting Players Take Center Stage , 2020, Neuron.

[4]  Lin Cheng,et al.  Single-cell landscape of bronchoalveolar immune cells in patients with COVID-19 , 2020, Nature Medicine.

[5]  Ruth R. Montgomery,et al.  Single-cell longitudinal analysis of SARS-CoV-2 infection in human airway epithelium , 2020, bioRxiv.

[6]  M. Hoffmann,et al.  A Multibasic Cleavage Site in the Spike Protein of SARS-CoV-2 Is Essential for Infection of Human Lung Cells , 2020, Molecular Cell.

[7]  L. Mao,et al.  Neurologic Manifestations of Hospitalized Patients With Coronavirus Disease 2019 in Wuhan, China. , 2020, JAMA neurology.

[8]  C. Lindskog,et al.  The protein expression profile of ACE2 in human tissues , 2020, bioRxiv.

[9]  P. Vollmar,et al.  Virological assessment of hospitalized patients with COVID-2019 , 2020, Nature.

[10]  Dan Zhang,et al.  Construction of a human cell landscape at single-cell level , 2020, Nature.

[11]  A. Walls,et al.  Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein , 2020, Cell.

[12]  G. Herrler,et al.  SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor , 2020, Cell.

[13]  B. Graham,et al.  Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation , 2020, Science.

[14]  B. Canard,et al.  The spike glycoprotein of the new coronavirus 2019-nCoV contains a furin-like cleavage site absent in CoV of the same clade , 2020, Antiviral Research.

[15]  Kai Zhao,et al.  A pneumonia outbreak associated with a new coronavirus of probable bat origin , 2020, Nature.

[16]  G. Gao,et al.  A Novel Coronavirus from Patients with Pneumonia in China, 2019 , 2020, The New England journal of medicine.

[17]  H. Matsunami,et al.  Single cell analysis of olfactory neurogenesis and differentiation in adult humans , 2020, Nature Neuroscience.

[18]  Victor M Corman,et al.  Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR , 2020, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[19]  Priti Kumar,et al.  A Positioning Device for the Placement of Mice During Intranasal siRNA Delivery to the Central Nervous System. , 2019, Journal of visualized experiments : JoVE.

[20]  Paul J. Hoffman,et al.  Comprehensive Integration of Single-Cell Data , 2018, Cell.

[21]  M. Hoffmann,et al.  Functional analysis of potential cleavage sites in the MERS-coronavirus spike protein , 2018, Scientific Reports.

[22]  A. Leitner,et al.  An Unbiased Screen for Human Cytomegalovirus Identifies Neuropilin-2 as a Central Viral Receptor , 2018, Cell.

[23]  J. Dye,et al.  NRP2 and CD63 Are Host Factors for Lujo Virus Cell Entry. , 2017, Cell host & microbe.

[24]  Kevin W. Eliceiri,et al.  ImageJ2: ImageJ for the next generation of scientific image data , 2017, BMC Bioinformatics.

[25]  Trevor Bedford,et al.  Multiplex PCR method for MinION and Illumina sequencing of Zika and other virus genomes directly from clinical samples , 2017, Nature Protocols.

[26]  E. Kieff,et al.  Neuropilin 1 is an entry factor that promotes EBV infection of nasopharyngeal epithelial cells , 2015, Nature Communications.

[27]  Erkki Ruoslahti,et al.  Etchable plasmonic nanoparticle probes to image and quantify cellular internalization , 2014, Nature materials.

[28]  C. Ruhrberg,et al.  Neuropilin Regulation of Angiogenesis, Arteriogenesis, and Vascular Permeability , 2014, Microcirculation.

[29]  Björn Usadel,et al.  Trimmomatic: a flexible trimmer for Illumina sequence data , 2014, Bioinform..

[30]  G. Whittaker,et al.  A Novel Activation Mechanism of Avian Influenza Virus H9N2 by Furin , 2013, Journal of Virology.

[31]  D. Sikkema,et al.  Development of an enzymatic assay for the detection of neutralizing antibodies against therapeutic angiotensin-converting enzyme 2 (ACE2). , 2013, Journal of immunological methods.

[32]  A. Wilm,et al.  LoFreq: a sequence-quality aware, ultra-sensitive variant caller for uncovering cell-population heterogeneity from high-throughput sequencing datasets , 2012, Nucleic acids research.

[33]  E. Ruoslahti,et al.  C-end rule peptides mediate neuropilin-1-dependent cell, vascular, and tissue penetration , 2009, Proceedings of the National Academy of Sciences.

[34]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[35]  P. van Endert,et al.  Neuropilin-1 Is Involved in Human T-Cell Lymphotropic Virus Type 1 Entry , 2006, Journal of Virology.

[36]  A. Scheffold,et al.  Single-cell analysis of the murine chemokines MIP-1α, MIP-1β, RANTES and ATAC/lymphotactin by flow cytometry , 2003 .

[37]  T. Kitsukawa,et al.  Developmentally regulated expression of a cell surface protein, neuropilin, in the mouse nervous system. , 1996, Journal of neurobiology.