The inactivation of the Niemann Pick C1 cholesterol transporter restricts SARS-CoV-2 entry into host cells by decreasing ACE2 abundance at the plasma membrane
Abstract Background The Niemann Pick C1 (NPC1) protein is an intracellular cholesterol transporter located in the late endosome/lysosome (LE/Ly) that is involved in the mobilization of endocytosed cholesterol. Loss-of-function mutations in the NPC1 gene lead to the accumulation of cholesterol and sp...
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BMC
2024-12-01
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| Series: | Cell & Bioscience |
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| Online Access: | https://doi.org/10.1186/s13578-024-01331-4 |
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| author | Piergiorgio La Rosa Jessica Tiberi Enrico Palermo Roberta Stefanelli Sofia Maria Luigia Tiano Sonia Canterini Mirko Cortese John Hiscott Maria Teresa Fiorenza |
| author_facet | Piergiorgio La Rosa Jessica Tiberi Enrico Palermo Roberta Stefanelli Sofia Maria Luigia Tiano Sonia Canterini Mirko Cortese John Hiscott Maria Teresa Fiorenza |
| author_sort | Piergiorgio La Rosa |
| collection | DOAJ |
| description | Abstract Background The Niemann Pick C1 (NPC1) protein is an intracellular cholesterol transporter located in the late endosome/lysosome (LE/Ly) that is involved in the mobilization of endocytosed cholesterol. Loss-of-function mutations in the NPC1 gene lead to the accumulation of cholesterol and sphingolipids in LE/Ly, resulting in severe fatal NPC1 disease. Cellular alterations associated with NPC1 inactivation affect both the integrity of lipid rafts and the endocytic pathway. Because the angiotensin-converting enzyme 2 (ACE2) and type 2 serine transmembrane protease (TMPRSS2), interactors of the SARS-CoV-2 Spike protein also localize to lipid rafts, we sought to investigate the hypothesis that NPC1 inactivation would generate an intrinsically unfavorable barrier to SARS-CoV-2 entry. Results In this study, we show that inhibition of the cholesterol transporter activity of NPC1 in cells that express both ACE2 and TMPRSS2, considerably reduces SARS-CoV-2 infectivity, evaluated as early as 4 h post-infection. Mechanistically, treatment with NPC1 specific inhibitor U18666A relocalizes ACE2 from the plasma membrane to the autophagosomal/lysosomal compartment, thereby reducing SARS-CoV-2 entry into treated cells. Reduction of viral entry was observed for both fully infectious SARS-CoV-2 virus and with a pseudotyped VSV-Spike-GFP virus. For instance, U18666A-treated Caco-2 cells infected with the pseudotyped VSV-Spike-GFP showed a > threefold and > 40-fold reduction in virus titer when infectivity was measured at 4 h or 24 h post-infection, respectively. A similar effect was observed in CRISP/R-Cas9-edited Caco-2 cells, which were even more resistant to SARS-CoV-2 infection as indicated by a 97% reduction of viral titers. Conclusion Overall, this study provides compelling evidence that the inhibition of NPC1 cholesterol transporter activity generates a cellular environment that hinders SARS-CoV-2 entry. ACE2 depletion from the plasma membrane appears to play a major role as limiting factor for viral entry. |
| format | Article |
| id | doaj-art-ee4df504b129455c9e01b1a609987f84 |
| institution | DOAJ |
| issn | 2045-3701 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | BMC |
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| series | Cell & Bioscience |
| spelling | doaj-art-ee4df504b129455c9e01b1a609987f842025-08-20T02:40:18ZengBMCCell & Bioscience2045-37012024-12-0114111610.1186/s13578-024-01331-4The inactivation of the Niemann Pick C1 cholesterol transporter restricts SARS-CoV-2 entry into host cells by decreasing ACE2 abundance at the plasma membranePiergiorgio La Rosa0Jessica Tiberi1Enrico Palermo2Roberta Stefanelli3Sofia Maria Luigia Tiano4Sonia Canterini5Mirko Cortese6John Hiscott7Maria Teresa Fiorenza8Division of Neuroscience, Dept. of Psychology, University La SapienzaDivision of Neuroscience, Dept. of Psychology, University La SapienzaIstituto Pasteur Italia-Cenci Bolognetti FoundationDivision of Neuroscience, Dept. of Psychology, University La SapienzaTelethon Institute of Genetics and Medicine, TIGEMDivision of Neuroscience, Dept. of Psychology, University La SapienzaTelethon Institute of Genetics and Medicine, TIGEMIstituto Pasteur Italia-Cenci Bolognetti FoundationDivision of Neuroscience, Dept. of Psychology, University La SapienzaAbstract Background The Niemann Pick C1 (NPC1) protein is an intracellular cholesterol transporter located in the late endosome/lysosome (LE/Ly) that is involved in the mobilization of endocytosed cholesterol. Loss-of-function mutations in the NPC1 gene lead to the accumulation of cholesterol and sphingolipids in LE/Ly, resulting in severe fatal NPC1 disease. Cellular alterations associated with NPC1 inactivation affect both the integrity of lipid rafts and the endocytic pathway. Because the angiotensin-converting enzyme 2 (ACE2) and type 2 serine transmembrane protease (TMPRSS2), interactors of the SARS-CoV-2 Spike protein also localize to lipid rafts, we sought to investigate the hypothesis that NPC1 inactivation would generate an intrinsically unfavorable barrier to SARS-CoV-2 entry. Results In this study, we show that inhibition of the cholesterol transporter activity of NPC1 in cells that express both ACE2 and TMPRSS2, considerably reduces SARS-CoV-2 infectivity, evaluated as early as 4 h post-infection. Mechanistically, treatment with NPC1 specific inhibitor U18666A relocalizes ACE2 from the plasma membrane to the autophagosomal/lysosomal compartment, thereby reducing SARS-CoV-2 entry into treated cells. Reduction of viral entry was observed for both fully infectious SARS-CoV-2 virus and with a pseudotyped VSV-Spike-GFP virus. For instance, U18666A-treated Caco-2 cells infected with the pseudotyped VSV-Spike-GFP showed a > threefold and > 40-fold reduction in virus titer when infectivity was measured at 4 h or 24 h post-infection, respectively. A similar effect was observed in CRISP/R-Cas9-edited Caco-2 cells, which were even more resistant to SARS-CoV-2 infection as indicated by a 97% reduction of viral titers. Conclusion Overall, this study provides compelling evidence that the inhibition of NPC1 cholesterol transporter activity generates a cellular environment that hinders SARS-CoV-2 entry. ACE2 depletion from the plasma membrane appears to play a major role as limiting factor for viral entry.https://doi.org/10.1186/s13578-024-01331-4Spike-ACE2 interactionACE2Virus entryLipid dyshomeostasis |
| spellingShingle | Piergiorgio La Rosa Jessica Tiberi Enrico Palermo Roberta Stefanelli Sofia Maria Luigia Tiano Sonia Canterini Mirko Cortese John Hiscott Maria Teresa Fiorenza The inactivation of the Niemann Pick C1 cholesterol transporter restricts SARS-CoV-2 entry into host cells by decreasing ACE2 abundance at the plasma membrane Cell & Bioscience Spike-ACE2 interaction ACE2 Virus entry Lipid dyshomeostasis |
| title | The inactivation of the Niemann Pick C1 cholesterol transporter restricts SARS-CoV-2 entry into host cells by decreasing ACE2 abundance at the plasma membrane |
| title_full | The inactivation of the Niemann Pick C1 cholesterol transporter restricts SARS-CoV-2 entry into host cells by decreasing ACE2 abundance at the plasma membrane |
| title_fullStr | The inactivation of the Niemann Pick C1 cholesterol transporter restricts SARS-CoV-2 entry into host cells by decreasing ACE2 abundance at the plasma membrane |
| title_full_unstemmed | The inactivation of the Niemann Pick C1 cholesterol transporter restricts SARS-CoV-2 entry into host cells by decreasing ACE2 abundance at the plasma membrane |
| title_short | The inactivation of the Niemann Pick C1 cholesterol transporter restricts SARS-CoV-2 entry into host cells by decreasing ACE2 abundance at the plasma membrane |
| title_sort | inactivation of the niemann pick c1 cholesterol transporter restricts sars cov 2 entry into host cells by decreasing ace2 abundance at the plasma membrane |
| topic | Spike-ACE2 interaction ACE2 Virus entry Lipid dyshomeostasis |
| url | https://doi.org/10.1186/s13578-024-01331-4 |
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