Identification of NPC1 as a novel SARS-CoV-2 intracellular target

García-Dorival, Isabel; Cuesta-Geijo, Miguel Ángel; Barrado-Gil, Lucía; Galindo, Inmaculada; Urquiza, Jesús; Puerto, Ana del; Gil, Carmen; Campillo, Nuria E.; Martínez, Ana María; Alonso, Covadonga

doi:10.1101/2020.12.19.423584

Cited by 3 publications

(6 citation statements)

References 46 publications

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“…Further elucidation of the latter and how it is modulated by viruses thus presents as important aspect of focus. After cholesterol-dependent steps in endocytosis, shuttling of the lipid within virus-containing endosomes further is important for the uncoating, and therefore for final cellular entry, of enveloped viruses, such as members of the Filoviridae family, IAV, HIV, SARS-CoV-2, and Alphaviruses ( Schroeder, 2010 ; Carette et al, 2011 ; Côté et al, 2011 ; Garcia-Dorival et al, 2020 ; Martyna et al, 2020 ; Sousa et al, 2020 ). This process is reported to be mediated via cholesterol-shuttling proteins, such as NPC1.…”

Section: Endosomal Cholesterol Trafficking As Shared Feature In Viral Infectionsmentioning

confidence: 99%

Endosomal Cholesterol in Viral Infections – A Common Denominator?

Glitscher

Hildt

2021

Front. Physiol.

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Cholesterol has gained tremendous attention as an essential lipid in the life cycle of virtually all viruses. These seem to have developed manifold strategies to modulate the cholesterol metabolism to the side of lipid uptake and de novo synthesis. In turn, affecting the cholesterol homeostasis has emerged as novel broad-spectrum antiviral strategy. On the other hand, the innate immune system is similarly regulated by the lipid and stimulated by its derivatives. This certainly requires attention in the design of antiviral strategies aiming to decrease cellular cholesterol, as evidence accumulates that withdrawal of cholesterol hampers innate immunity. Secondly, there are exceptions to the rule of the abovementioned virus-induced metabolic shift toward cholesterol anabolism. It therefore is of interest to dissect underlying regulatory mechanisms, which we aimed for in this minireview. We further collected evidence for intracellular cholesterol concentrations being less important in viral life cycles as compared to the spatial distribution of the lipid. Various routes of cholesterol trafficking were found to be hijacked in viral infections with respect to organelle-endosome contact sites mediating cholesterol shuttling. Thus, re-distribution of cellular cholesterol in the context of viral infections requires more attention in ongoing research. As a final aim, a pan-antiviral treatment could be found just within the transport and re-adjustment of local cholesterol concentrations. Thus, we aimed to emphasize the importance of the regulatory roles the endosomal system fulfils herein and hope to stimulate research in this field.

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Section: Endosomal Cholesterol Trafficking As Shared Feature In Viral Infectionsmentioning

confidence: 99%

Endosomal Cholesterol in Viral Infections – A Common Denominator?

Glitscher

Hildt

2021

Front. Physiol.

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“…Recently, a positive interaction between NPC1 and SARS-CoV-2 Nucleoprotein was reported (22). Consequently, NPC1 appears to be relevant in a variety of RNA or DNA viral families.…”

Section: Discussionmentioning

confidence: 99%

“…These compounds act either by binding to NPC1 sterol sensing domain (SSD) or by blocking EBOV Glycoprotein (GP) GP-NPC1 interaction, respectively. This indicates that NPC1 and intact cholesterol transport are pivotal for a successful infection (21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32).…”

mentioning

confidence: 99%

Endosomal proteins NPC1 and NPC2 at African swine fever virus entry/fusion

Alonso

Cuesta-Geijo

Urquiza

et al. 2021

Preprint

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African swine fever virus (ASFV) infectious cycle starts with the viral adsorption and entry into the host cell. The virus is internalized via clathrin/dynamin mediated endocytosis and macropinocytosis. As several other viruses, ASF virion is then internalized and incorporated into the endocytic pathway. Endosomal maturation entails luminal acidification and the lowering of pH acting on the multi-layered virion structure dissolves the outer capsid. Upon decapsidation, the inner viral membrane is exposed to interact with the limiting membrane of the late endosome for fusion. Egress from endosome is related to cholesterol efflux, but it remains an intriguing process albeit essential for infection, specifically for the viral nucleic acid exit to the cytoplasm for replication. ASFV proteins E248R and E199L, with structural homology to the VACV proteins of the fusion complex, seem to have similar functions in ASFV. A direct interaction between these ASFV proteins with the cholesterol transporter protein NPC1 (Niemann-Pick C type 1) was observed, which was also shared by the E248R homologous protein L1R of VACV. Binding occurs between the transmembrane domain of E248R with the loop C of NPC1 at the same domain than EBOV binding site. These interactions suggest that these ASFV proteins are crucial for membrane fusion. CRISPR NPC1 KO Vero cells lacking NPC1 protein that were resistant to EBOV, reduced ASFV infection levels significantly. Reductions on ASFV infectivity and replication in NPC1 KO cells were accompanied by lesser viral factories of smaller size and lacking the typical cohesive morphology between endosomes and viral proteins. We observed a compensatory effect in NPC1 KO cells, elevating NPC2 levels while silencing NPC2 in Vero cells with shRNA, also reduced ASFV infection. Our findings pave the way to understand the role of these proteins at the membrane viral fusion step for several viruses.

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“…82,83 A recent study identified an interaction between SARS-CoV-2 nucleoprotein and NPC1, suggesting NPC1 to be a facilitator for the endocytosis of SARS-CoV-2. 26 U18666A, an inhibitor of NPC1, 84 significantly inhibited the infection and replication of SARS-CoV-2 in Vero E6 and Calu-3 cells. 27 This anti-SARS-CoV-2 activity was associated with increased cholesterol storage and reduced acidification in endo/lysosomes, consequences of blocking NPC1.…”

Section: Npc1mentioning

confidence: 94%

“…In this review, we focus on the proteins ACE2, TMPRSS2, CTSL, PIKfyve, NPC1, CLTC, EGFR, PPIA, eEF1A, and S100A8. These proteins are of interest because their roles in SARS-CoV-2 infection, replication, or pathogenesis have been well-characterized (e.g., ACE2, TMPRSS2, 24 CTSL, 25 NPC1, 26,27 CLTC, 28 and S100A8 29 ) by cell or animal models, or their inhibitors have shown promising anti-SARS-CoV-2 activities in preclinical studies (e.g., PIKfyve, 12,30 EGFR, 8,31 PPIA, [32][33][34] and eEF1A 11 ). These host proteins can be categorized according to their putative roles in the SARS-CoV-2 life cycle: (a) cell membrane attachment (ACE2), 24 (b) membrane fusion (TMPRSS2), 24 (c) endocytosis (CTSL, 25 PIKfyve, 12,30 NPC1, 26,27 and CLTC 28 ), (d) replication and translation of the viral genome (EGFR, 8,31 PPIA, [32][33][34] and eEF1A 11 ), and (e) immune response (S100A8 29 ; Figure 1).…”

Section: Host Protein Selectionmentioning

confidence: 99%

Tissue‐ and cell‐expression of druggable host proteins provide insights into repurposing drugs for COVID‐19

Xue

Wang

et al. 2022

Clinical Translational Sci

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Several human host proteins play important roles in the lifecycle of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2). Many drugs targeting these host proteins have been investigated as potential therapeutics for coronavirus disease 2019 (COVID‐19). The tissue‐specific expressions of selected host proteins were summarized using proteomics data retrieved from the Human Protein Atlas, ProteomicsDB, Human Proteome Map databases, and a clinical COVID‐19 study. Protein expression features in different cell lines were summarized based on recent proteomics studies. The half‐maximal effective concentration or half‐maximal inhibitory concentration values were collected from in vitro studies. The pharmacokinetic data were mainly from studies in healthy subjects or non‐COVID‐19 patients. Considerable tissue‐specific expression patterns were observed for several host proteins. ACE2 expression in the lungs was significantly lower than in many other tissues (e.g., the kidneys and intestines); TMPRSS2 expression in the lungs was significantly lower than in other tissues (e.g., the prostate and intestines). The expression levels of endocytosis‐associated proteins CTSL, CLTC, NPC1, and PIKfyve in the lungs were comparable to or higher than most other tissues. TMPRSS2 expression was markedly different between cell lines, which could be associated with the cell‐dependent antiviral activities of several drugs. Drug delivery receptor ICAM1 and CTSB were expressed at a higher level in the lungs than in other tissues. In conclusion, the cell‐ and tissue‐specific proteomics data could help interpret the in vitro antiviral activities of host‐directed drugs in various cells and aid the transition of the in vitro findings to clinical research to develop safe and effective therapeutics for COVID‐19.

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Identification of NPC1 as a novel SARS-CoV-2 intracellular target

Cited by 3 publications

References 46 publications

Endosomal Cholesterol in Viral Infections – A Common Denominator?

Endosomal Cholesterol in Viral Infections – A Common Denominator?

Endosomal proteins NPC1 and NPC2 at African swine fever virus entry/fusion

Tissue‐ and cell‐expression of druggable host proteins provide insights into repurposing drugs for COVID‐19

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