Potential of targeting host cell calcium dynamics to curtail SARS-CoV-2 infection and COVID-19 pathogenesis

Sultan, Farina; Ahuja, Kriti; Motiani, Rajender K.

doi:10.1016/j.ceca.2022.102637

Cited by 11 publications

(10 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…It is apparent from these data that this approach could not assay specific FP residues’ sensitivity to intracellular calcium. Disruption of physiological concentrations of calcium is known to impact the fusion and infectivity of SARS-CoV-1 and SARS-CoV-2 (28). We propose that calcium chelation via BAPTA-AM treatment disrupted endosomal calcium concentrations causing a major reduction in endosomal fusion of WT pseudoparticles, as SARS-CoV-1 enters VeroE6 cells primarily via the endosomal/lysosomal pathway.…”

Section: Discussionmentioning

confidence: 99%

A mechanistic understanding of the modes of Ca ion binding to the SARS-CoV-1 fusion peptide and their role in the dynamics of host membrane penetration

Carten

Bidon

Khelashvili

et al. 2022

Preprint

View full text Add to dashboard Cite

Viral envelope fusion with the host cell membrane is dependent on a specific viral fusion peptide (FP) or loop, which becomes exposed during virus entry to drive the process of membrane fusion. In coronaviruses, the FP is a highly conserved domain that sits in the center of spike protein and in SARS-CoV, is adjacent to the S2’ proteolytic cleavage site. This peptide contains a hydrophobic LLF motif, as well as several conserved negatively charged amino acids that interact with Ca2+ ions to promote membrane fusion. In this work we perform a systematic mutagenesis study of the negatively charged amino acids within the SARS-CoV fusion peptide (FP1/FP2) and combine this with molecular dynamics simulations to define the membrane interactions that regulate virus infectivity. We show that the E801/D802 amino acid pair in the SARS-CoV FP is predicted to bind to one Ca2+ ion to promote FP-membrane interaction, with a second Ca2+ ion likely pairing residue D812 with either E821 or D825. The D812/D821 residue pair promotes membrane interaction, whereas the D821/D825 is inhibitory to membrane insertion. Taken together, our results demonstrate the dynamic nature of the coronavirus FP region that likely facilitates its interactions with and insertion into the host cell membrane.Author SummaryCoronaviruses have reemerged as a highly pathogenic virus family through the rise of SARS-CoV, MERS-CoV, and more recently, SARS-CoV-2. As more transmissible variants of SARS-CoV-2 arise, it is imperative that we understand the mechanisms of CoV viral entry to enable the development of effective therapeutics. Recent reviews have suggested the repurposing of FDA-approved calcium channel blockers to treat infection by coronaviruses; however, calcium’s method of action on viral-host cell fusion events is unknown. We have found that increased calcium availability leads to increased viral infection across the CoV family, suggesting that calcium is involved in mediating the interaction between the viral fusion peptide and the host cell membrane. As such, we hypothesize that the highly conserved fusion peptide interacts directly with calcium and this interaction is required for viral entry and infection. Through mutagenesis studies of specific negatively charged residues in the fusion peptide, we have identified residues that impact viral infectivity. We have also compared the infectivity of wild-type and mutant CoV pseudoparticles in calcium-rich or -depleted environments using chelating drugs. Our data mirrors the residue coordination observed SARS-CoV-2, as both between SARS-CoV and SARS-CoV-2 FPs bind to two calcium ions. These results demonstrate the importance of Ca2+ for CoV FP function during viral entry and opens the possibility of utilizing FDA-approved calcium-blocking drugs as a treatment for COVID-19.

show abstract

Section: Discussionmentioning

confidence: 99%

A mechanistic understanding of the modes of Ca ion binding to the SARS-CoV-1 fusion peptide and their role in the dynamics of host membrane penetration

Carten

Bidon

Khelashvili

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…PERK activation also results in enhanced ATF4 levels that activate a transcriptional response program that can either be adaptive (through autophagy induction) or apoptotic (through production of CHOP and activation of pro-apoptotic proteins) [28,29] One of the most critical contributors to ER stress induction is dysregulated ER Ca 2+ homeostasis. It is important to note that perturbations in ER Ca 2+ signaling can lead to a variety of viral pathogenesis and therefore, it is emerging as a potential therapeutic target [30,31]. Since MTP induces ER stress response, we examined if it can modulate ER Ca 2+ signaling.…”

Section: Phenothiazines Are Mtor Independent Autophagy Inducersmentioning

confidence: 99%

Methotrimeprazine exerts antiviral and neuroprotective effects in Japanese encephalitis virus infection through activation of adaptive ER stress and autophagy

Prajapat

Mishra

Khera

et al. 2023

Preprint

View full text Add to dashboard Cite

Japanese encephalitis virus (JEV) is the leading global cause of virus-induced encephalitis. Its pathogenesis is driven by a combination of neuronal cell death and neuroinflammation. We hypothesized that pharmacological upregulation of autophagy could exert a neuroprotective antiviral effect, and tested a panel of forty-two FDA-approved drugs that were shown to induce autophagy. Four drugs were tested in the JE mouse model based on in vitro protective effects on neuronal cell death, inhibition of viral replication, and anti-inflammatory effects in microglial cells. The antipsychotic phenothiazines Methotrimeprazine (MTP) and Trifluoperazine (TFP) showed a significant survival benefit with reduced virus titers in the brain, prevention of blood-brain barrier (BBB) breach, and inhibition of neuroinflammation. Both drugs were potent mTOR-independent autophagy flux inducers. Mechanistically MTP inhibited SERCA channel functioning, thereby resulting in rise in cytosolic calcium levels, and induction of a unique adaptive ER stress response. In virus infected drug treated cells, there was a strong transcriptional downregulation of type I interferon and interferon-stimulated genes and upregulation of cholesterol metabolic pathway genes. The drugs exerted an autophagy-dependent antiviral effect at the level of JEV protein translation/replication complex formation in diverse cell types. Inhibition of inflammatory cytokine/chemokine release from mouse microglial cells was partly autophagy-dependent. Our study suggests that MTP exerts a combined antiviral and anti-inflammatory effect in JEV infection, and has therapeutic potential to be repurposed for JE treatment.

show abstract

“…Accordingly, the E protein is a viroporin, normally used by RNA viruses to subvert the host cell ion homeostasis to facilitate viral infection [ 30 , 31 ], forming a cation-selective voltage-dependent channel across the ERGIC membrane [ 32 , 33 ]. The cation permeability of the E protein can change based on ions concentration (K + , Na + , Ca 2+ and Mg 2+ ) [ 32 , 34 ], composition of the cell membrane [ 33 , 35 ] and pH [ 30 ]. SARS-CoV-2 has also been shown to perturb host cell Ca 2+ homeostasis highlighting the critical role of Ca 2+ for infection and pathogenesis [ 34 ].…”

Section: Introductionmentioning

confidence: 99%

“…The cation permeability of the E protein can change based on ions concentration (K + , Na + , Ca 2+ and Mg 2+ ) [ 32 , 34 ], composition of the cell membrane [ 33 , 35 ] and pH [ 30 ]. SARS-CoV-2 has also been shown to perturb host cell Ca 2+ homeostasis highlighting the critical role of Ca 2+ for infection and pathogenesis [ 34 ]. Therefore, ion leakage via the E protein might play an important role in the development of respiratory inflammation via inflammasome activation [ 29 , 36 ].…”

Section: Introductionmentioning

confidence: 99%

Perturbation of the host cell Ca2+ homeostasis and ER-mitochondria contact sites by the SARS-CoV-2 structural proteins E and M

et al. 2023

View full text Add to dashboard Cite

Coronavirus disease (COVID-19) is a contagious respiratory disease caused by the SARS-CoV-2 virus. The clinical phenotypes are variable, ranging from spontaneous recovery to serious illness and death. On March 2020, a global COVID-19 pandemic was declared by the World Health Organization (WHO). As of February 2023, almost 670 million cases and 6,8 million deaths have been confirmed worldwide. Coronaviruses, including SARS-CoV-2, contain a single-stranded RNA genome enclosed in a viral capsid consisting of four structural proteins: the nucleocapsid (N) protein, in the ribonucleoprotein core, the spike (S) protein, the envelope (E) protein, and the membrane (M) protein, embedded in the surface envelope. In particular, the E protein is a poorly characterized viroporin with high identity amongst all the β-coronaviruses (SARS-CoV-2, SARS-CoV, MERS-CoV, HCoV-OC43) and a low mutation rate. Here, we focused our attention on the study of SARS-CoV-2 E and M proteins, and we found a general perturbation of the host cell calcium (Ca2+) homeostasis and a selective rearrangement of the interorganelle contact sites. In vitro and in vivo biochemical analyses revealed that the binding of specific nanobodies to soluble regions of SARS-CoV-2 E protein reversed the observed phenotypes, suggesting that the E protein might be an important therapeutic candidate not only for vaccine development, but also for the clinical management of COVID designing drug regimens that, so far, are very limited.

show abstract

Potential of targeting host cell calcium dynamics to curtail SARS-CoV-2 infection and COVID-19 pathogenesis

Cited by 11 publications

References 60 publications

A mechanistic understanding of the modes of Ca ion binding to the SARS-CoV-1 fusion peptide and their role in the dynamics of host membrane penetration

A mechanistic understanding of the modes of Ca ion binding to the SARS-CoV-1 fusion peptide and their role in the dynamics of host membrane penetration

Methotrimeprazine exerts antiviral and neuroprotective effects in Japanese encephalitis virus infection through activation of adaptive ER stress and autophagy

Perturbation of the host cell Ca2+ homeostasis and ER-mitochondria contact sites by the SARS-CoV-2 structural proteins E and M

Contact Info

Product

Resources

About