Systematic analysis of SARS-CoV-2 infection of an ACE2-negative human airway cell

Puray-Chavez, Maritza; LaPak, Kyle M.; Schrank, Travis P.; Elliott, Jennifer L; Bhatt, Dhaval P.; Agajanian, Megan; Jasuja, Ria; Lawson, Dana Q.; Davis, Keanu; Rothlauf, Paul W.; Liu, Zhuoming; Jo, Heejoon; Lee, Nakyung; Tenneti, Kasyap; Eschbach, Jenna E.; Mugisha, Christian Shema; Cousins, Emily; Cloer, Erica W.; Vuong, Hung R.; VanBlargan, Laura A.; Bailey, Adam L.; Gilchuk, Pavlo; Crowe, James E.; Diamond, Michael S.; Hayes, D. Neil; Whelan, Sean P. J.; Horani, Amjad; Brody, Steven L.; Goldfarb, Dennis; Major, Michael B.

doi:10.1016/j.celrep.2021.109364

Cited by 116 publications

(128 citation statements)

References 127 publications

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“…Moreover, SARS-CoV-2 infects the brain, and viral RNA has been detected in immune cells such as neutrophils, macrophages, T/B cells, and NK cells (Grant et al, 2021;Ren et al, 2021), whereas ACE2 is barely detected in these tissues or cells (Figure S1 in Supporting Information). Second, a recent report showed that an ACE2-null lung adenocarcinoma cell is highly permissive to SARS-CoV-2 (Puray- Chavez et al, 2021), indicating that SARS-CoV-2 could leverage an alternative receptor for its entry. Third, a cell surface protein, AXL, has recently been reported to facilitate SARS-CoV-2 entry independently of ACE2 (Wang et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

Genome-wide CRISPR activation screen identifies candidate receptors for SARS-CoV-2 entry

Zhu

Liu

Zhou

et al. 2021

Sci. China Life Sci.

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The outbreak of coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 has created a global health crisis. SARS-CoV-2 infects varieties of tissues where the known receptor ACE2 is low or almost absent, suggesting the existence of alternative viral entry pathways. Here, we performed a genome-wide barcoded-CRISPRa screen to identify novel host factors that enable SARS-CoV-2 infection. Beyond known host proteins, i.e., ACE2, TMPRSS2, and NRP1, we identified multiple host components, among which LDLRAD3, TMEM30A, and CLEC4G were confirmed as functional receptors for SARS-CoV-2. All these membrane proteins bind directly to spike’s N-terminal domain (NTD). Their essential and physiological roles have been confirmed in either neuron or liver cells. In particular, LDLRAD3 and CLEC4G mediate SARS-CoV-2 entry and infection in an ACE2-independent fashion. The identification of the novel receptors and entry mechanisms could advance our understanding of the multiorgan tropism of SARS-CoV-2, and may shed light on the development of COVID-19 countermeasures. Supporting Information The supporting information is available online at 10.1007/s11427-021-1990-5. The supporting materials are published as submitted, without typesetting or editing. The responsibility for scientific accuracy and content remains entirely with the authors.

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Section: Introductionmentioning

confidence: 99%

Genome-wide CRISPR activation screen identifies candidate receptors for SARS-CoV-2 entry

Zhu

Liu

Zhou

et al. 2021

Sci. China Life Sci.

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“…There is evidence to suggest that SARS-CoV-2 uses plasma membrane fusion as the default pathway but can use endosomal fusion if the plasma membrane protease, TMPRSS2, is not available; hence, the micro-environment is important in dictating the entry pathway [14]. However, it has been found that infection of ACE2-deficient lung cells depends on clathrin-mediated endocytosis and endosomal cathepsin L, indicating that endosomal fusion may well be the major entry pathway in a subset of cell types [15]. Endosomal fusion is preceded by receptor-mediated endocytosis and trafficking to an acidic compartment to trigger fusion [5].…”

Section: Introductionmentioning

confidence: 99%

Kite-Shaped Molecules Block SARS-CoV-2 Cell Entry at a Post-Attachment Step

Chan

Shafi

Ford

2021

Viruses

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Anti-viral small molecules are currently lacking for treating coronavirus infection. The long development timescales for such drugs are a major problem, but could be shortened by repurposing existing drugs. We therefore screened a small library of FDA-approved compounds for potential severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) antivirals using a pseudovirus system that allows a sensitive read-out of infectivity. A group of structurally-related compounds, showing moderate inhibitory activity with IC50 values in the 2–5 μM range, were identified. Further studies demonstrated that these “kite-shaped” molecules were surprisingly specific for SARS-CoV-1 and SARS-CoV-2 and that they acted early in the entry steps of the viral infectious cycle, but did not affect virus attachment to the cells. Moreover, the compounds were able to prevent infection in both kidney- and lung-derived human cell lines. The structural homology of the hits allowed the production of a well-defined pharmacophore that was found to be highly accurate in predicting the anti-viral activity of the compounds in the screen. We discuss the prospects of repurposing these existing drugs for treating current and future coronavirus outbreaks.

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“…Heat-inactivated plasma from vaccinated or control mice were serially diluted in DMEM with 2% FBS, 1% Penicillin-Streptomycin, and 10 mM Non-Essential Amino Acids (Gibco; mixture of glycine, l -alanine, l -asparagine, l -aspartic acid, l -glutamic acid, l -proline, & l -serine)), and subsequently incubated with 400 plaque-forming units of SARS-CoV-2 virus (strain nCov/Washington/1/2020, provided by the National Biocontainment Laboratory, Galveston TX, USA) for 1 h at 37 °C. These mixtures were then applied to Vero-E6 cells, which endogenously express ACE2 at high levels and show ACE-2 dependent SARS-CoV-2 infection [ 6 , 46 ]. These mixtures were maintained with the Vero-E6 cells until >90% cell death occurred in the “no serum" control condition (about 4–5 days).…”

Section: Methodsmentioning

confidence: 99%

Generation of potent cellular and humoral immunity against SARS-CoV-2 antigens via conjugation to a polymeric glyco-adjuvant

Gray¹,

Raczy²,

Briquez³

et al. 2021

Biomaterials

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The SARS-CoV-2 virus has caused an unprecedented global crisis, and curtailing its spread requires an effective vaccine which elicits a diverse and robust immune response. We have previously shown that vaccines made of a polymeric glyco-adjuvant conjugated to an antigen were effective in triggering such a response in other disease models and hypothesized that the technology could be adapted to create an effective vaccine against SARS-CoV-2. The core of the vaccine platform is the copolymer p(Man-TLR7), composed of monomers with pendant mannose or a toll-like receptor 7 (TLR7) agonist. Thus, p(Man-TLR7) is designed to target relevant antigen-presenting cells (APCs) via mannose-binding receptors and then activate TLR7 upon endocytosis. The p(Man-TLR7) construct is amenable to conjugation to protein antigens such as the Spike protein of SARS-CoV-2, yielding Spike-p(Man-TLR7). Here, we demonstrate Spike-p(Man-TLR7) vaccination elicits robust antigen-specific cellular and humoral responses in mice. In adult and elderly wild-type mice, vaccination with Spike-p(Man-TLR7) generates high and long-lasting titers of anti-Spike IgGs, with neutralizing titers exceeding levels in convalescent human serum. Interestingly, adsorbing Spike-p(Man-TLR7) to the depot-forming adjuvant alum amplified the broadly neutralizing humoral responses to levels matching those in mice vaccinated with formulations based off of clinically-approved adjuvants. Additionally, we observed an increase in germinal center B cells, antigen-specific antibody secreting cells, activated T follicular helper cells, and polyfunctional Th1-cytokine producing CD4 + and CD8 + T cells. We conclude that Spike-p(Man-TLR7) is an attractive, next-generation subunit vaccine candidate, capable of inducing durable and robust antibody and T cell responses.

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Systematic analysis of SARS-CoV-2 infection of an ACE2-negative human airway cell

Cited by 116 publications

References 127 publications

Genome-wide CRISPR activation screen identifies candidate receptors for SARS-CoV-2 entry

Genome-wide CRISPR activation screen identifies candidate receptors for SARS-CoV-2 entry

Kite-Shaped Molecules Block SARS-CoV-2 Cell Entry at a Post-Attachment Step

Generation of potent cellular and humoral immunity against SARS-CoV-2 antigens via conjugation to a polymeric glyco-adjuvant

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