{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T02:21:08Z","timestamp":1789438868631,"version":"build-2803163510"},"reference-count":22,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2025,4,14]],"date-time":"2025-04-14T00:00:00Z","timestamp":1744588800000},"content-version":"vor","delay-in-days":13,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2025,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Lenacapavir (LEN) is a potent, first\u2010in\u2010class long\u2010acting HIV\u20101 capsid (CA) inhibitor, approved for treatment of people with multi\u2010drug resistant HIV in combination with other antiretrovirals. In clinical studies with LEN, the CA resistance\u2010associated mutations (RAMs) M66I, Q67H\/K\/N, K70H\/N\/R\/S, N74D\/H\/K, A105S\/T, and T107A\/C\/N\/S were observed. Phenotypic analyses of these CA mutants were possible in single cycle (SC) PhenoSense Gag\u2010Pro assays; however, most CA mutants exhibited severely impaired replication capacity (RC), rendering phenotyping with multicycle (MC) MT\u20102 cytopathic assays unsuccessful for many. Here, we developed and optimized a novel MC phenotyping assay using a Rev\u2010dependent HIV reporter cell line, Rev\u2010CEM\u2010Luc\/GFP (RevLun assay), to further characterize these replication\u2010impaired LEN\u2010resistant HIV CA mutants. HIV Gag\u2010Protease fragments from patients' plasma samples with CA RAMs and associated site\u2010directed mutants were cloned into the pXXLAI HIV molecular clone, and replicative viral supernatants were evaluated in MT\u20102 and RevLun MC assays, with readouts of cytopathic effect and reporter gene expression, respectively. Viruses were also evaluated in the SC assay. We successfully phenotyped CA mutants in RevLun that were noninfectious in MT\u20102 assay, including clinical isolates containing M66I in various genetic contexts and combinations of LEN RAMs. LEN susceptibility in the RevLun MC assay aligned with data in the SC PhenoSense Gag\u2010Pro assay and MC MT\u20102 assay when available. All viruses with LEN RAMs remained sensitive to other HIV drug classes. Using a sensitive HIV\u2010dependent reporter system enhanced our ability to assess the phenotypes of viruses with low RC, allowing for further investigation into LEN resistance and CA RAMs.<\/jats:p>","DOI":"10.1002\/jmv.70340","type":"journal-article","created":{"date-parts":[[2025,4,14]],"date-time":"2025-04-14T05:57:01Z","timestamp":1744610221000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Phenotypic Characterization of Replication\u2010Impaired Lenacapavir\u2010Resistant HIV Clinical Isolates"],"prefix":"10.1002","volume":"97","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0659-0614","authenticated-orcid":false,"given":"Sally","family":"Demirdjian","sequence":"first","affiliation":[{"name":"HIV Clinical Virology Gilead Sciences Inc. Foster City California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vidula","family":"Naik","sequence":"additional","affiliation":[{"name":"HIV Clinical Virology Gilead Sciences Inc. Foster City California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0911-4237","authenticated-orcid":false,"given":"Nicolas","family":"Margot","sequence":"additional","affiliation":[{"name":"HIV Clinical Virology Gilead Sciences Inc. Foster City California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brie","family":"Falkard","sequence":"additional","affiliation":[{"name":"HIV Clinical Virology Gilead Sciences Inc. Foster City California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian","family":"Callebaut","sequence":"additional","affiliation":[{"name":"HIV Clinical Virology Gilead Sciences Inc. Foster City California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,4,14]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2443-1"},{"key":"e_1_2_9_3_1","unstructured":"SUNLENCA and Gilead Sciences Inc. SUNLENCA\u00ae (lenacapavir) injection for subcutaneous use. US Prescribing Information.Foster City CA.July.2024."},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2407001"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkab503"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/S2352-3018(23)00113-3"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/S2352-3018(22)00291-0"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2115542"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/ciae423"},{"key":"e_1_2_9_10_1","unstructured":"N.Margot V.Jogiraju L.VanderVeen et al. \u201cResistance Analysis of Long\u2010Acting Lenacapavir in Heavily Treatment\u2010Experienced People with HIV after 104 Weeks of Treatment. [Oral: PS8 O4]. The 19th European AIDS Conference \u201d in2023"},{"key":"e_1_2_9_11_1","unstructured":"D.Hagins E.Koenig R.Safran et al. Long\u2010Acting Lenacapavir in a Combination Regimen for Treatment Na\u00efve PWH: Week 80 [Poster 522]. Conference on Retroviruses and Opportunistic Infections (CROI) \u201d in2023. Seattle WA."},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jiac364"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1177\/13596535231220754"},{"key":"e_1_2_9_14_1","unstructured":"L. A.VanderVeen Resistance Analysis of Long\u2010Acting Lenacapavir in People With HIV Who are Treatment\u2010Na\u00efve After 80 Weeks of Treatment [Poster14]. European Meeting on HIV & Hepatitis \u201d in2023. Rome Italy."},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.2174\/157016207781024018"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.41.12.2781"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.02057-20"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.4.920-928.2000"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01183-10"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.3390\/v13010125"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002896"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkaa060"},{"key":"e_1_2_9_23_1","unstructured":"C. F.Kelley \u201cTwice\u2010Yearly Lenacapavir for HIV Prevention in Cisgender Gay Bisexual and Other Men Transgender Women Transgender Men and Gender\u2010Nonbinary People who Have Sex With Partners Assigned Male at Birth:Interim Analysis Results From the PURPOSE 2 Study [Oral Presentation] \u201d in5th HIV Research for Prevention Conference (HIVR4P).2024.Lima Peru."}],"container-title":["Journal of Medical Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmv.70340","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T09:03:15Z","timestamp":1764666195000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jmv.70340"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4]]},"references-count":22,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2025,4]]}},"alternative-id":["10.1002\/jmv.70340"],"URL":"https:\/\/doi.org\/10.1002\/jmv.70340","archive":["Portico"],"relation":{},"ISSN":["0146-6615","1096-9071"],"issn-type":[{"value":"0146-6615","type":"print"},{"value":"1096-9071","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4]]},"assertion":[{"value":"2024-11-20","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-03-28","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-04-14","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70340"}}