Heparan Sulfate Facilitates Spike Protein-Mediated SARS-CoV-2 Host Cell Invasion and Contributes to Increased Infection of SARS-CoV-2 G614 Mutant and in Lung Cancer

Abstract

The severe acute respiratory syndrome (SARS)-like coronavirus disease (COVID-19) is caused by SARS-CoV-2 and has been a serious threat to global public health with limited treatment. Cellular heparan sulfate (HS) has been found to bind SARS-CoV-2 spike protein (SV2-S) and co-operate with cell surface receptor angiotensin-converting enzyme 2 (ACE2) to mediate SARS-CoV-2 infection of host cells. In this study, we determined that host cell surface SV2-S binding depends on and correlates with host cell surface HS expression. This binding is required for SARS-Cov-2 virus to infect host cells and can be blocked by heparin lyase, HS antagonist surfen, heparin, and heparin derivatives. The binding of heparin/HS to SV2-S is mainly determined by its overall sulfation with potential, minor contribution of specific SV2-S binding motifs. The higher binding affinity of SV2-S G614 mutant to heparin and upregulated HS expression may be one of the mechanisms underlying the higher infectivity of the SARS-CoV-2 G614 variant and the high vulnerability of lung cancer patients to SARS-CoV-2 infection, respectively. The higher host cell infection by SARS-CoV-2 G614 variant pseudovirus and the increased infection caused by upregulated HS expression both can be effectively blocked by heparin lyase and heparin, and possibly surfen and heparin derivatives too. Our findings support blocking HS-SV2-S interaction may provide one addition to achieve effective prevention and/treatment of COVID-19.

GlycoGrip: Cell Surface-Inspired Universal Sensor for Betacoronaviruses

“…Cellular HS is incredibly heterogeneous in both uronic acid identity and degree of sulfation. 23 , 26 , 27 , 29 , 56 − 59 To better match HS considered in our LFSA design and testing (where HS was purchased from Sigma and reported to be only 5–7% sulfation by mass), we constructed 6- O -sulfated heparan sulfate, from here on referred to as our H6S model. Although this model does not capture the full heterogeneity of cellular HS, it does better reflect the low sulfation rate of HS considered experimentally in this work.…”

“…H6S was chosen as a defined sequence for modeling and docking as it captures the 5–7% sulfation rate reported by Sigma-Aldrich while also capturing potentially important interactions facilitated by the 6-O sulfation position reported by several groups. 23 , 26 , 27 , 29 , 56 , 59 As docking of long chain polysaccharides to large protein structures is combinatorially intractable, we modeled small dimer and tetramer structures of each sulfated polysaccharide. Dimeric sulfated polysaccharides were modeled with the intention of capturing highly localized interactions, while tetrameric sulfated polysaccharides were modeled with the intention of capturing steric hindrance effects encountered with larger substrates.…”

A glycosaminoglycan microarray identifies the binding of SARS‐CoV‐2 spike protein to chondroitin sulfate E

“…The GAG microarrays were then applied to screen for GAGs recognized by the SARS‐CoV‐2 S protein, which consists of S1 and S2 subunits [ 3 , 4 ]. The binding capacity of the SARS‐CoV‐2 S protein to various GAGs has been analyzed using various binding analyses such as surface plasmon resonance (SPR), pulldown assay, flow cytometry, cell‐based ELISA, and viral entry assays (Table S2 ) [ 5 , 8 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 ]. The SARS‐CoV‐2 S protein (monomeric form) showed the highest signal intensity to HEP‐BSA (Fig.…”

“…The addition of HEP and its derivatives was demonstrated to inhibit SARS‐CoV‐2 infection [ 5 ]. The binding capacity of the SARS‐CoV‐2 S protein to various GAGs has also been reported using various binding analyses such as SPR, pulldown assay, flow cytometry, cell‐based ELISA, and viral entry assays (Table S2 ) [ 5 , 8 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 ].…”

Structural characteristics of Heparan sulfate required for the binding with the virus processing Enzyme Furin

“…Inspired by these facts, we hypothesize that the Furin is also a heparin/heparan sulfate binding protein. The most recent discovery by ourselves and other groups that cell surface heparan sulfate is a potential co-receptor for SARS-CoV-2 infection further stimulates us to test the hypothesis [ 11 – 14 ].…”