Hydroxychloroquine blocks SARS-CoV-2 entry into the endocytic pathway in mammalian cell culture

DOI record: { "DOI": "10.1038/s42003-022-03841-8", "ISSN": [ "2399-3642" ], "URL": "http://dx.doi.org/10.1038/s42003-022-03841-8", "abstract": "<jats:title>Abstract</jats:title><jats:p>Hydroxychloroquine (HCQ), a drug used to treat lupus and malaria, was proposed as a treatment for SARS-coronavirus-2 (SARS-CoV-2) infection, albeit with controversy. In vitro, HCQ effectively inhibits viral entry, but its use in the clinic has been hampered by conflicting results. A better understanding of HCQ’s mechanism of actions in vitro is needed. Recently, anesthetics were shown to disrupt ordered clusters of monosialotetrahexosylganglioside1 (GM1) lipid. These same lipid clusters recruit the SARS-CoV-2 surface receptor angiotensin converting enzyme 2 (ACE2) to endocytic lipids, away from phosphatidylinositol 4,5 bisphosphate (PIP<jats:sub>2</jats:sub>) clusters. Here we employed super-resolution imaging of cultured mammalian cells (VeroE6, A549, H1793, and HEK293T) to show HCQ directly perturbs clustering of ACE2 receptor with both endocytic lipids and PIP<jats:sub>2</jats:sub> clusters. In elevated (high) cholesterol, HCQ moves ACE2 nanoscopic distances away from endocytic lipids. In cells with resting (low) cholesterol, ACE2 primarily associates with PIP<jats:sub>2</jats:sub> clusters, and HCQ moves ACE2 away from PIP<jats:sub>2</jats:sub> clusters—erythromycin has a similar effect. We conclude HCQ inhibits viral entry through two distinct mechanisms in high and low tissue cholesterol and does so prior to inhibiting cathepsin-L. HCQ clinical trials and animal studies will need to account for tissue cholesterol levels when evaluating dosing and efficacy.</jats:p>", "alternative-id": [ "3841" ], "article-number": "958", "assertion": [ { "group": { "label": "Article History", "name": "ArticleHistory" }, "label": "Received", "name": "received", "order": 1, "value": "8 November 2021" }, { "group": { "label": "Article History", "name": "ArticleHistory" }, "label": "Accepted", "name": "accepted", "order": 2, "value": "12 August 2022" }, { "group": { "label": "Article History", "name": "ArticleHistory" }, "label": "First Online", "name": "first_online", "order": 3, "value": "14 September 2022" }, { "group": { "label": "Competing interests", "name": "EthicsHeading" }, "name": "Ethics", "order": 1, "value": "The authors declare no competing interests." } ], "author": [ { "ORCID": "http://orcid.org/0000-0003-0665-5086", "affiliation": [], "authenticated-orcid": false, "family": "Yuan", "given": "Zixuan", "sequence": "first" }, { "affiliation": [], "family": "Pavel", "given": "Mahmud Arif", "sequence": "additional" }, { "affiliation": [], "family": "Wang", "given": "Hao", "sequence": "additional" }, { "ORCID": "http://orcid.org/0000-0003-4313-9187", "affiliation": [], "authenticated-orcid": false, "family": "Kwachukwu", "given": "Jerome C.", "sequence": "additional" }, { "affiliation": [], "family": "Mediouni", "given": "Sonia", "sequence": "additional" }, { "affiliation": [], "family": "Jablonski", "given": "Joseph Anthony", "sequence": "additional" }, { "ORCID": "http://orcid.org/0000-0003-2917-7574", "affiliation": [], "authenticated-orcid": false, "family": "Nettles", "given": "Kendall W.", "sequence": "additional" }, { "affiliation": [], "family": "Reddy", "given": "Chakravarthy B.", "sequence": "additional" }, { "affiliation": [], "family": "Valente", "given": "Susana T.", "sequence": "additional" }, { "ORCID": "http://orcid.org/0000-0003-0086-9753", "affiliation": [], "authenticated-orcid": false, "family": "Hansen", "given": "Scott B.", "sequence": "additional" } ], "container-title": "Communications Biology", "container-title-short": "Commun Biol", "content-domain": { "crossmark-restriction": false, "domain": [ "link.springer.com" ] }, "created": { "date-parts": [ [ 2022, 9, 15 ] ], "date-time": "2022-09-15T09:29:55Z", "timestamp": 1663234195000 }, "deposited": { "date-parts": [ [ 2022, 9, 15 ] ], "date-time": "2022-09-15T09:34:34Z", "timestamp": 1663234474000 }, "funder": [ { "DOI": "10.13039/100000065", "award": [ "R01NS112534" ], "doi-asserted-by": "publisher", "name": "U.S. Department of Health & Human Services | NIH | National Institute of Neurological Disorders and Stroke" }, { "DOI": "10.13039/100000005", "award": [ "W81XWH1810782" ], "doi-asserted-by": "publisher", "name": "U.S. Department of Defense" } ], "indexed": { "date-parts": [ [ 2022, 9, 16 ] ], "date-time": "2022-09-16T06:23:53Z", "timestamp": 1663309433452 }, "is-referenced-by-count": 0, "issue": "1", "issued": { "date-parts": [ [ 2022, 9, 14 ] ] }, "journal-issue": { "issue": "1", "published-online": { "date-parts": [ [ 2022, 12 ] ] } }, "language": "en", "license": [ { "URL": "https://creativecommons.org/licenses/by/4.0", "content-version": "tdm", "delay-in-days": 0, "start": { "date-parts": [ [ 2022, 9, 14 ] ], "date-time": "2022-09-14T00:00:00Z", "timestamp": 1663113600000 } }, { "URL": "https://creativecommons.org/licenses/by/4.0", "content-version": "vor", "delay-in-days": 0, "start": { "date-parts": [ [ 2022, 9, 14 ] ], "date-time": "2022-09-14T00:00:00Z", "timestamp": 1663113600000 } } ], "link": [ { "URL": "https://www.nature.com/articles/s42003-022-03841-8.pdf", "content-type": "application/pdf", "content-version": "vor", "intended-application": "text-mining" }, { "URL": "https://www.nature.com/articles/s42003-022-03841-8", "content-type": "text/html", "content-version": "vor", "intended-application": "text-mining" }, { "URL": "https://www.nature.com/articles/s42003-022-03841-8.pdf", "content-type": "application/pdf", "content-version": "vor", "intended-application": "similarity-checking" } ], "member": "297", "original-title": [], "prefix": "10.1038", "published": { "date-parts": [ [ 2022, 9, 14 ] ] }, "published-online": { "date-parts": [ [ 2022, 9, 14 ] ] }, "publisher": "Springer Science and Business Media LLC", "reference": [ { "DOI": "10.1016/j.ijsu.2020.02.034", "author": "C Sohrabi", "doi-asserted-by": "publisher", "first-page": "71", "journal-title": "Int. 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