SARS-CoV-2 Infects Human ACE2-Negative Endothelial Cells through an αvβ3 Integrin-Mediated Endocytosis Even in the Presence of Vaccine-Elicited Neutralizing Antibodies

Bugatti, Antonella; Filippini, Federica; Bardelli, Marta; Zani, Alberto; Chiodelli, Paola; Messali, Serena; Caruso, Arnaldo; Caccuri, Francesca

doi:10.3390/v14040705

Cited by 28 publications

(32 citation statements)

References 61 publications

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“…Of reports investigating integrin-spike interactions, α 5 β 1 has been proposed as a receptor of interest, in part due to its functional association with ACE2 (75,76), the ability of β1-selective integrin antagonists to reduce SARS-Co-V2 invasion (35,38,39), and observed α 5 β 1 integrin-S1-RBD interactions by ELISA (35) and SPR (79) assays. Some reports have also implicated α v β 3 integrins in viral entry (37,80), while others found no effects of integrin antagonists on viral invasion (34). In the present study, we compared effects of β 1 - and β 3 -integrin blocking antibodies on cell attachment to RBD using a well-characterized fibroblast cell line that expresses both functional α 5 β 1 and α v β 3 integrin receptors (41,81).…”

Section: Discussionmentioning

confidence: 99%

Receptor binding domain of SARS-CoV-2 is a functional αv-integrin agonist

Norris

Pan

Hocking

2022

Preprint

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Among the novel mutations distinguishing SARS-CoV-2 from similar respiratory coronaviruses is a K403R substitution in the receptor-binding domain (RBD) of the viral spike (S) protein within its S1 region. This amino acid substitution occurs near the angiotensin-converting enzyme 2 (ACE2)-binding interface and gives rise to a canonical RGD adhesion motif that is often found in native extracellular matrix proteins, including fibronectin. In the present study, the ability of recombinant S1-RBD to bind to cell surface integrins and trigger downstream signaling pathways was assessed and compared to RGD-containing, integrin-binding fragments of fibronectin. S1-RBD supported adhesion of both fibronectin-null mouse embryonic fibroblasts as well as primary human small airway epithelial cells. Cell adhesion to S1-RBD was cation- and RGD-dependent, and was inhibited by blocking antibodies against αv and β3, but not α5 or β1, integrins. Similarly, direct binding of S1-RBD to recombinant human αvβ3 and αvβ6 integrins, but not α5β1 integrins, was observed by surface plasmon resonance. Adhesion to S1-RBD initiated cell spreading, focal adhesion formation, and actin stress fiber organization to a similar extent as fibronectin. Moreover, S1-RBD stimulated tyrosine phosphorylation of the adhesion mediators FAK, Src, and paxillin, Akt activation, and supported cell proliferation. Together, these data demonstrate that the RGD sequence within S1-RBD can function as an αv-selective integrin agonist. This study provides evidence that cell surface αv-containing integrins can respond functionally to spike protein and raise the possibility that S1-mediated dysregulation of ECM dynamics may contribute to the pathogenesis and/or post-acute sequelae of SARS-CoV-2 infection.

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Section: Discussionmentioning

confidence: 99%

Receptor binding domain of SARS-CoV-2 is a functional αv-integrin agonist

Norris

Pan

Hocking

2022

Preprint

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“…There are other several potential host entry factors on ECs as well as ACE2 (Fig. 1A ): neuropilin-1 (NRP1), which is a promoter of virus entry in the presence of ACE2 and TMPRESS2 [ 78 , 79 ], scavenger receptor B type 1 (SR-B1) [ 80 , 81 ] and extracellular vimentin [ 82 ], which facilitate ACE2-dependent virus entry; CD147 [ 83 ] and αvβ3 integrin [ 84 ], which mediate virus entry by endocytosis; and liver/lymph node-specific ICAM-3 grabbing nonintegrin (L-SIGN), which are highly expressed on human liver sinusoidal endothelial cells and serve as the virus receptor [ 85 ]. Histamine also contributes to the entry of spike protein into ECs [ 86 ].…”

Section: Mechanisms For Long Covid and Cardiovascular Damagementioning

confidence: 99%

Long COVID and hypertension-related disorders: a report from the Japanese Society of Hypertension Project Team on COVID-19

et al. 2022

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s The coronavirus disease 2019 (COVID-19) affects infected patients even after the acute phase and impairs their health and quality of life by causing a wide variety of symptoms, referred to as long COVID. Although the evidence is still insufficient, hypertension is suspected to be a potential risk factor for long COVID, and the occurrence of cardiovascular diseases seems to be a key facet of multiple conditions observed in long COVID. Nonetheless, there are few reports that comprehensively review the impacts of long COVID on hypertension and related disorders. As a sequel to our previous report in 2020 which reviewed the association of COVID-19 and hypertension, we summarize the possible influences of long COVID on hypertension-related organs, including the cardiovascular system, kidney, and endocrine system, as well as the pathophysiological mechanisms associated with the disorders in this review. Given that the clinical course of COVID-19 is highly affected by age and sex, we also review the impacts of these factors on long COVID. Lastly, we discuss areas of uncertainty and future directions, which may lead to better understanding and improved prognosis of clinical problems associated with COVID-19.

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“…1 hour later cells were infected with SARS-CoV-2 at a multiplicity of infection (MOI) of 0.01 and cultured for 72 hours. Four strains were used: 1) the human 2019-nCoV strain 2019-nCoV/Italy-INMI1, isolated in Italy (ex-China) from a sample collected on January 29, 2020, kindly provided by the Istituto Lazzaro Spallanzani, Rome, Italy [19], 2) GZ69 (genomic data are available at EBI under study accession n. PRJEB38101), 3) Delta variant B.1.167.2 and 4) Omicron BA.1 variant, the last three viral strains kindly provided by the Department of Molecular and Translational Medicine, Section of Microbiology and Virology, University of Brescia Medical School, Brescia, Italy [20][21][22][23]. The 2019-nCoV/Italy-INMI1 strain was used for initial experiments and for drug combination studies, GZ69 was used as confirmatory strain as it was the main circulating variant in Italy at the time experiments were performed.…”

Section: Antiviral Activity Of Drugsmentioning

confidence: 99%

Synergistic drug combinations designed to fully suppress SARS-CoV-2 in the lung of COVID-19 patients

Forni¹,

Poddesu²,

Cugia³

et al. 2022

PLoS ONE

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Despite new antivirals are being approved against SARS-CoV-2 they suffer from significant constraints and are not indicated for hospitalized patients, who are left with few antiviral options. Repurposed drugs have previously shown controversial clinical results and it remains difficult to understand why certain trials delivered positive results and other trials failed. Our manuscript contributes to explaining the puzzle: this might have been caused by a suboptimal drug exposure and, consequently, an incomplete virus suppression, also because the drugs have mostly been used as add-on monotherapies. As with other viruses (e.g., HIV and HCV) identifying synergistic combinations among such drugs could overcome monotherapy-related limitations. In a cell culture model for SARS-CoV-2 infection the following stringent criteria were adopted to assess drug combinations: 1) identify robust, synergistic antiviral activity with no increase in cytotoxicity, 2) identify the lowest drug concentration inhibiting the virus by 100% (LIC100) and 3) understand whether the LIC100 could be reached in the lung at clinically indicated drug doses. Among several combinations tested, remdesivir with either azithromycin or ivermectin synergistically increased the antiviral activity with no increase in cytotoxicity, improving the therapeutic index and lowering the LIC100 of every one of the drugs to levels that are expected to be achievable and maintained in the lung for a therapeutically relevant period of time. These results are consistent with recent clinical observations showing that intensive care unit admission was significantly delayed by the combination of AZI and RDV, but not by RDV alone, and could have immediate implications for the treatment of hospitalized patients with COVID-19 as the proposed “drug cocktails” should have antiviral activity against present and future SARS-CoV-2 variants without significant overlapping toxicity, while minimizing the onset of drug resistance. Our results also provide a validated methodology to help sort out which combination of drugs are most likely to be efficacious in vivo, based on their in vitro activity, potential synergy and PK profiles.

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SARS-CoV-2 Infects Human ACE2-Negative Endothelial Cells through an αvβ3 Integrin-Mediated Endocytosis Even in the Presence of Vaccine-Elicited Neutralizing Antibodies

Cited by 28 publications

References 61 publications

Receptor binding domain of SARS-CoV-2 is a functional αv-integrin agonist

Receptor binding domain of SARS-CoV-2 is a functional αv-integrin agonist

Long COVID and hypertension-related disorders: a report from the Japanese Society of Hypertension Project Team on COVID-19

Synergistic drug combinations designed to fully suppress SARS-CoV-2 in the lung of COVID-19 patients

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