CoChat
Sign Up
Cited by 240interferon and immune responsesSARS-CoV-2 and COVID-19 ResearchCOVID-19 Clinical Research Studies

Cholesterol 25-hydroxylase suppresses SARS-CoV-2 replication by blocking membrane fusion

Open access
Zang, Ruochen, Case, James Brett, Yutuc, Eylan, Ma, Xiucui, Shen, Sheng, Gomez Castro, Maria Florencia, Liu, Zhuoming, Zeng, Qiru +19 more·2020·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.2012197117PMC PMC7749331

Published version of a preprint

Open this paper in CoChat — free account

Abstract

Significance: The novel severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the etiological agent of coronavirus disease-2019 (COVID-19), has swept the world in unprecedented speed. In a few months, SARS-CoV-2 has infected millions of people and caused tens of thousands of deaths. There are no Food and Drug Administration-approved antivirals or vaccines yet available and clinical treatments are limited to supportive therapies that help alleviate the symptoms. Thus, there is an urgent need to identify effective antivirals as countermeasures before safe and effective vaccines are developed, tested, and then produced on a large scale. Our approach is to harness the germline-encoded interferon antiviral response to inhibit SARS-CoV-2 replication thereby limiting its pathogenicity.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1073/pnas.2012197117

Cites

5 of 5 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • 25-hydroxycholesterol confers broad antiviral activity by blocking entry, replication and production of infectious viruses of diverse enveloped viruses.

    “Previous reports have demonstrated that the natural cholesterol metabolite 25-hydroxycholesterol (25-HC), an immune modulator secreted by macrophages during the host response, confers broad antiviral activity by blocking entry, replication and production of infectious viruses of diverse enveloped viruses39–42”

    From Peroxisomal very long-chain fatty acid transport is targeted by herpesviruses and the antiviral host response (2022) · cites this paper as [42]

    The paper reports that 25HC has broad activity against enveloped viruses and shows that it inhibits SARS-CoV-2 entry and replication (Abstract, page not provided). It does not establish that 25HC blocks production of infectious virus: the authors say that effects on viral maturation, release, or progeny infectivity warrant further investigation (Abstract, page not provided). (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • 25-HC is effective in vitro against a wide range of viruses including SARS-CoV-2.

    “25-HC is effective in vitro against a wide range of viruses including SARS-CoV-242”

    From Peroxisomal very long-chain fatty acid transport is targeted by herpesviruses and the antiviral host response (2022) · cites this paper as [42]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 70

  1. [2]Zhou P., A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature 579, 270– 273 (2020). 32015507 PMC7095418
  2. [3]Donnelly R. P., Kotenko S. V., Interferon-lambda: A new addition to an old family. J. Interferon Cytokine Res. 30, 555– 564 (2010). 20712453 PMC2925029
  3. [4]Durbin J. E., Type I IFN modulates innate and specific antiviral immunity. J. Immunol. 164, 4220– 4228 (2000). 10754318
  4. [5]Schoggins J. W., Rice C. M., Interferon-stimulated genes and their antiviral effector functions. Curr. Opin. Virol. 1, 519– 525 (2011). 22328912 PMC3274382
  5. [6]Blanco-Melo D., Imbalanced hostresponse to SARS-CoV-2 drives development of COVID-19. Cell 181, 1036– 1045.e9 (2020). 32416070 PMC7227586
  6. [7]Lamers M. M., SARS-CoV-2 productively infects human gut enterocytes. Science 369, 50– 54 (2020). 32358202 PMC7199907
  7. [8]Ogando N. S., SARS-coronavirus-2 replication in Vero E6 cells: Replication kinetics, rapid adaptation and cytopathology. bioRxiv: 10.1101/2020.04.20.049924 (20 April 2020). PMC7654748 32568027 · DOI
  8. [9]Lokugamage K. G., Hage A., Schindewolf C., Rajsbaum R., Menachery V. D., SARS-CoV-2 is sensitive to type I interferon pretreatment. bioRxiv: 10.1101/2020.03.07.982264 (18 March 2020). · DOI
  9. [10]Stanifer M. L., Critical role of type III interferon in controlling SARS-CoV-2 infection in human intestinal epithelial cells. Cell Rep. 32, 107863 (2020). 32610043 PMC7303637
  10. [11]Vanderheiden A., Type I and type III IFN restrict SARS-CoV-2 infection of human airway epithelial cultures. bioRxiv: 10.1101/2020.05.19.105437 (20 May 2020). PMC7495371 32699094 · DOI
  11. [12]Dusheiko G., Side effects of alpha interferon in chronic hepatitis C. Hepatology 26(suppl. 1) 112S– 121S (1997). 9305675
  12. [13]Major J., Type I and III interferons disrupt lung epithelial repair during recovery from viral infection. Science 369, 712– 717 (2020). 32527928 PMC7292500
  13. [14]Case J. B., Neutralizing antibody and soluble ACE2 inhibition of a replication-competent VSV-SARS-CoV-2 and a clinical isolate of SARS-CoV-2. Cell Host Microbe 28, 475– 485.e5 (2020). 32735849 PMC7332453
  14. [15]Hoffmann M., SARS-CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell 181, 271– 280.e8 (2020). 32142651 PMC7102627
  15. [17]Walls A. C., Structure, function, and antigenicity of the SARS-CoV-2 spike glycoprotein. Cell 181, 281– 292.e6 (2020). 32155444 PMC7102599
  16. [18]Zang R., TMPRSS2 and TMPRSS4 promote SARS-CoV-2 infection of human small intestinal enterocytes. Sci. Immunol. 5, eabc3582 (2020). 32404436 PMC7285829
  17. [19]Loo Y. M., Gale M. Jr, Immune signaling by RIG-I-like receptors. Immunity 34, 680– 692 (2011). 21616437 PMC3177755
  18. [20]Pfaender S., LY6E impairs coronavirus fusion and confers immune control of viral disease. Nat. Microbiol. 5, 1330– 1339 (2020). 32704094 PMC7916999
  19. [21]Zhao X., LY6E restricts entry of human coronaviruses, including currently pandemic SARS-CoV-2. J. Virol. 94, e00562-20 (2020). 32641482 PMC7459569
  20. [22]Shi G., Opposing activities of IFITM proteins in SARS-CoV-2 infection. bioRxiv: 10.1101/2020.08.11.246678 (11 August 2020). PMC7744865 33270927 · DOI
  21. [23]Lund E. G., Kerr T. A., Sakai J., Li W. P., Russell D. W., cDNA cloning of mouse and human cholesterol 25-hydroxylases, polytopic membrane proteins that synthesize a potent oxysterol regulator of lipid metabolism. J. Biol. Chem. 273, 34316– 34327 (1998). 9852097
  22. [24]Hannedouche S., Oxysterols direct immune cell migration via EBI2. Nature 475, 524– 527 (2011). 21796212 PMC4297623
  23. [25]Schoggins J. W., Interferon-stimulated genes: What do they all do? Annu. Rev. Virol. 6, 567– 584 (2019). 31283436
  24. [26]Blanc M., The transcription factor STAT-1 couples macrophage synthesis of 25-hydroxycholesterol to the interferon antiviral response. Immunity 38, 106– 118 (2013). 23273843 PMC3556782
  25. [27]Zhang Y., Cholesterol 25-hydroxylase negatively regulates porcine intestinal coronavirus replication by the production of 25-hydroxycholesterol. Vet. Microbiol. 231, 129– 138 (2019). 30955800 PMC7117535
  26. [28]Liu S. Y., Interferon-inducible cholesterol-25-hydroxylase broadly inhibits viral entry by production of 25-hydroxycholesterol. Immunity 38, 92– 105 (2013). 23273844 PMC3698975
  27. [29]Sharma A., Human iPSC-derived cardiomyocytes are susceptible to SARS-CoV-2 infection. Cell Rep Med 1, 100052 (2020). 32835305 PMC7323681
  28. [30]Chen Y., Interferon-inducible cholesterol-25-hydroxylase inhibits hepatitis C virus replication via distinct mechanisms. Sci. Rep. 4, 7242 (2014). 25467815 PMC4252895
  29. [31]Matsuyama S., Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells. Proc. Natl. Acad. Sci. U.S.A. 117, 7001– 7003 (2020). 32165541 PMC7132130
  30. [32]Bertram S., TMPRSS2 activates the human coronavirus 229E for cathepsin-independent host cell entry and is expressed in viral target cells in the respiratory epithelium. J. Virol. 87, 6150– 6160 (2013). 23536651 PMC3648130
  31. [33]Huang I. C., Distinct patterns of IFITM-mediated restriction of filoviruses, SARS coronavirus, and influenza A virus. PLoS Pathog. 7, e1001258 (2011). 21253575 PMC3017121
  32. [35]Podbilewicz B., Virus and cell fusion mechanisms. Annu. Rev. Cell Dev. Biol. 30, 111– 139 (2014). 25000995
  33. [36]Xia S., Inhibition of SARS-CoV-2 (previously 2019-nCoV) infection by a highly potent pan-coronavirus fusion inhibitor targeting its spike protein that harbors a high capacity to mediate membrane fusion. Cell Res. 30, 343– 355 (2020). 32231345 PMC7104723
  34. [37]Li K., IFITM proteins restrict viral membrane hemifusion. PLoS Pathog. 9, e1003124 (2013). 23358889 PMC3554583
  35. [38]Sengupta P., A lipid-based partitioning mechanism for selective incorporation of proteins into membranes of HIV particles. Nat. Cell Biol. 21, 452– 461 (2019). 30936472
  36. [39]Kwon H. J., Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol. Cell 137, 1213– 1224 (2009). 19563754 PMC2739658
  37. [40]Carstea E. D., Niemann-pick C1 disease gene: Homology to mediators of cholesterol homeostasis. Science 277, 228– 231 (1997). 9211849
  38. [42]Richardson R. B., A CRISPR screen identifies IFI6 as an ER-resident interferon effector that blocks flavivirus replication. Nat. Microbiol. 3, 1214– 1223 (2018). 30224801 PMC6202210
  39. [44]Schoggins J. W., A diverse range of gene products are effectors of the type I interferon antiviral response. Nature 472, 481– 485 (2011). 21478870 PMC3409588
  40. [45]Ganser-Pornillos B. K., Pornillos O., Restriction of HIV-1 and other retroviruses by TRIM5. Nat. Rev. Microbiol. 17, 546– 556 (2019). 31312031 PMC6858284
  41. [46]Chen D., GILT restricts the cellular entry mediated by the envelope glycoproteins of SARS-CoV, Ebola virus and Lassa fever virus. Emerg. Microbes Infect. 8, 1511– 1523 (2019). 31631785 PMC6818130
  42. [47]Nchioua R., The zinc finger antiviral protein restricts SARS-CoV-2. bioRxiv: 10.1101/2020.06.04.134379 (8 June 2020). · DOI
  43. [48]Anggakusuma, Interferon-inducible cholesterol-25-hydroxylase restricts hepatitis C virus replication through blockage of membranous web formation. Hepatology 62, 702– 714 (2015). 25999047
  44. [49]Doms A., Sanabria T., Hansen J. N., Altan-Bonnet N., Holm G. H., 25-Hydroxycholesterol production by the cholesterol-25-hydroxylase interferon-stimulated gene restricts mammalian reovirus infection. J. Virol. 92, e01047-18 (2018). 29950420 PMC6146694
  45. [50]Shawli G. T., Adeyemi O. O., Stonehouse N. J., Herod M. R., The oxysterol 25-hydroxycholesterol inhibits replication of murine norovirus. Viruses 11, 97 (2019). PMC6409565 30682775
  46. [51]Civra A., 25-Hydroxycholesterol and 27-hydroxycholesterol inhibit human rotavirus infection by sequestering viral particles into late endosomes. Redox Biol. 19, 318– 330 (2018). 30212801 PMC6138790
  47. [52]Arias C. F., Silva-Ayala D., López S., Rotavirus entry: A deep journey into the cell with several exits. J. Virol. 89, 890– 893 (2015). 25378490 PMC4300671
  48. [53]Wang J., Cholesterol 25-hydroxylase acts as a host restriction factor on pseudorabies virus replication. J. Gen. Virol. 98, 1467– 1476 (2017). 28631596
  49. [54]Civra A., Inhibition of pathogenic non-enveloped viruses by 25-hydroxycholesterol and 27-hydroxycholesterol. Sci. Rep. 4, 7487 (2014). 25501851 PMC4265783
  50. [55]Shrivastava-Ranjan P., 25-Hydroxycholesterol inhibition of Lassa virus infection through aberrant GP1 glycosylation. MBio 7, e01808-16 (2016). 27999160 PMC5181775
  51. [56]Xiang Y., Identification of cholesterol 25-hydroxylase as a novel host restriction factor and a part of the primary innate immune responses against hepatitis C virus infection. J. Virol. 89, 6805– 6816 (2015). 25903345 PMC4468479
  52. [57]Kang Y. L., Inhibition of PIKfyve kinase prevents infection by Zaire ebolavirus and SARS-CoV-2. Proc. Natl. Acad. Sci. U.S.A. 117, 20803– 20813 (2020). 32764148 PMC7456157
  53. [58]Zu S., 25-Hydroxycholesterol is a potent SARS-CoV-2 inhibitor. Cell Res. 30, 1043– 1045 (2020). 32811977 PMC7431750
  54. [59]Wang S., Cholesterol 25-Hydroxylase inhibits SARS-CoV-2 and other coronaviruses by depleting membrane cholesterol. EMBO J. 39, e106057 (2020). 32944968 PMC7537045
  55. [60]Carette J. E., Ebola virus entry requires the cholesterol transporter Niemann-Pick C1. Nature 477, 340– 343 (2011). 21866103 PMC3175325
  56. [61]Jin Z., Structure of Mpro from SARS-CoV-2 and discovery of its inhibitors. Nature 582, 289– 293 (2020). 32272481
  57. [62]Zhang L., Crystal structure of SARS-CoV-2 main protease provides a basis for design of improved α-ketoamide inhibitors. Science 368, 409– 412 (2020). 32198291 PMC7164518
  58. [63]Gao Y., Structure of the RNA-dependent RNA polymerase from COVID-19 virus. Science 368, 779– 782 (2020). 32277040 PMC7164392
  59. [64]Wolf M., Deal E. M., Greenberg H. B., Rhesus rotavirus trafficking during entry into MA104 cells is restricted to the early endosome compartment. J. Virol. 86, 4009– 4013 (2012). 22278225 PMC3302545
  60. [65]Gordon D. E., A SARS-CoV-2-human protein-protein interaction map reveals drug targets and potential drug-repurposing. bioRxiv: 10.1101/2020.03.22.002386 (27 March 2020). · DOI
  61. [66]Simmons G., Characterization of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike glycoprotein-mediated viral entry. Proc. Natl. Acad. Sci. U.S.A. 101, 4240– 4245 (2004). 15010527 PMC384725
  62. [67]Kanai Y., Entirely plasmid-based reverse genetics system for rotaviruses. Proc. Natl. Acad. Sci. U.S.A. 114, 2349– 2354 (2017). 28137864 PMC5338561
  63. [68]Li B., Drebrin restricts rotavirus entry by inhibiting dynamin-mediated endocytosis. Proc. Natl. Acad. Sci. U.S.A. 114, E3642– E3651 (2017). 28416666 PMC5422808
  64. [69]Ding S., Rotavirus VP3 targets MAVS for degradation to inhibit type III interferon expression in intestinal epithelial cells. eLife 7, e39494 (2018). 30460894 PMC6289572
  65. [70]Crick P. J., Quantitative charge-tags for sterol and oxysterol analysis. Clin. Chem. 61, 400– 411 (2015). 25512642
  66. [71]Campeau E., A versatile viral system for expression and depletion of proteins in mammalian cells. PLoS One 4, e6529 (2009). 19657394 PMC2717805
  67. [72]Shang J., Structural basis of receptor recognition by SARS-CoV-2. Nature 581, 221– 224 (2020). 32225175 PMC7328981
  68. [73]Bolen C. R., Ding S., Robek M. D., Kleinstein S. H., Dynamic expression profiling of type I and type III Interferon-stimulated hepatocytes reveals a stable hierarchy of gene expression. Hepatology 59, 1262– 1272 (2013). PMC3938553 23929627
  69. [74]Ding S., STAG2 deficiency induces interferon responses via cGAS-STING pathway and restricts virus infection. Nat. Commun. 9, 1485 (2018). 29662124 PMC5902600
  70. [75]Ding S., Khoury-Hanold W., Iwasaki A., Robek M. D., Epigenetic reprogramming of the type III interferon response potentiates antiviral activity and suppresses tumor growth. PLoS Biol. 12, e1001758 (2014). 24409098 PMC3883642

Cited by

Sorted by citations

Authors
Zang, R., Case, J. B., Yutuc, E., Ma, X., Shen, S., Gomez Castro, M. F., Liu, Z., Zeng, Q., Zhao, H., Son, J., Rothlauf, P. W., Kreutzberger, A. J. B., Hou, G., Zhang, H., Bose, S., Wang, X., Vahey, M. D., Mani, K., Griffiths, W. J., Kirchhausen, T., Fremont, D. H., Guo, H., Diwan, A., Wang, Y., Diamond, M. S., Whelan, S. P. J., & Ding, S.
Affiliations
  • Washington University in St. Louis (US)
  • Swansea University (GB)
  • Indiana University Indianapolis (US)
  • Department of Molecular Microbiology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110;
  • Harvard University (US)
  • Boston Children's Hospital (US)
  • Autonomous Therapeutics, Inc., New York, NY 10013;
  • Ocean University of China (CN)
  • Program in Cellular and Molecular Medicine, Boston Children’s Hospital, Boston, MA 02115;
Corresponding author
Ding, Siyuan
ORCID
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
2020-11-25
Volume
117
Issue
50
Pages
32105-32113
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.2012197117
PMC
PMC7749331
OpenAlex
W3109188198
Cited by
240
References
5
Citations by year
2026: 19 · 2025: 16 · 2024: 28 · 2023: 48 · 2022: 60 · 2021: 62 · 2020: 7
FWCI
15.59
Type
journal article
Language
en
Open access
Licence
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Digital Commons@Becker (Washington University School of Medicine) (submittedVersion, cc-by)
  • IUScholarWorks (Indiana University) (submittedVersion, cc-by)
Funders
  • RCUK | Biotechnology and Biological Sciences Research Council (BB/L001942/1)
  • HHS | NIH | National Institute of Allergy and Infectious Diseases (R00 AI135031, R01 AI150796, R01 AI127828, 75N93019C00062, R37 AI059371)
  • HHS | NIH | National Institute of Diabetes and Digestive and Kidney Diseases (P30 DK052574)
  • DOD | Defense Advanced Research Projects Agency (HR001117S0019)
  • Helen Hay Whitney Foundation (postdoctoral fellowship)
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › interferon and immune responses
  • Health Sciences › Medicine › Infectious Diseases › SARS-CoV-2 and COVID-19 Research
  • Health Sciences › Medicine › Infectious Diseases › COVID-19 Clinical Research Studies
Keywords
cholesterol 25-hydroxylase, 25-hydroxycholesterol, SARS-CoV-2, viral fusion, spike protein, interferon-stimulated genes, antiviral mechanism, vesicular stomatitis virus, late endosomes, cholesterol export, antiviral drug discovery
MeSH terms
COVID-19, SARS-CoV-2, COVID-19 Drug Treatment, Antiviral Agents, Humans, Hydroxycholesterols, Interferons, Membrane Fusion, Endosomes, Virus Replication, Severe acute respiratory syndrome-related coronavirus, Virus Internalization, Spike Glycoprotein, Coronavirus

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review