{"doi":"10.1128/jvi.01510-23","title":"Adaptation of SARS-CoV-2 to ACE2 <sub>H353K</sub> mice reveals new spike residues that drive mouse infection","abstract":"ABSTRACT The Coronavirus Disease 2019 (COVID-19) pandemic continues to cause extraordinary loss of life and economic damage. Animal models of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) infection are needed to better understand disease pathogenesis and evaluate preventive measures and therapies. While mice are widely used to model human disease, mouse angiotensin converting enzyme 2 (ACE2) does not bind the ancestral SARS-CoV-2 spike protein to mediate viral entry. To overcome this limitation, we “humanized” mouse Ace2 using CRISPR gene editing to introduce a single amino acid substitution, H353K, predicted to facilitate S protein binding. While H353K knockin Ace2 (mACE2 H353K ) mice supported SARS-CoV-2 infection and replication, they exhibited minimal disease manifestations. Following 30 serial passages of ancestral SARS-CoV-2 in mACE2 H353K mice, we generated and cloned a more virulent virus. A single isolate (SARS2 MA-H353K ) was prepared for detailed studies. In 7–11-month-old mACE2 H353K mice, a 10 4 PFU inocula resulted in diffuse alveolar disease manifested as edema, hyaline membrane formation, and interstitial cellular infiltration/thickening. Unexpectedly, the mouse-adapted virus also infected standard BALB/c and C57BL/6 mice and caused severe disease. The mouse-adapted virus acquired five new missense mutations including two in spike (K417E, Q493K), one each in nsp4, nsp9, and M and a single nucleotide change in the 5′ untranslated region. The Q493K spike mutation arose early in serial passage and is predicted to provide affinity-enhancing molecular interactions with mACE2 and further increase the stability and affinity to the receptor. This new model and mouse-adapted virus will be useful to evaluate COVID-19 disease and prophylactic and therapeutic interventions. IMPORTANCE We developed a new mouse model with a humanized angiotensin converting enzyme 2 (ACE2) locus that preserves native regulatory elements. A single point mutation in mouse ACE2 (H353K) was sufficient to confer in vivo infection with ancestral severe acute respiratory syndrome-coronavirus-2 virus. Through in vivo serial passage, a virulent mouse-adapted strain was obtained. In aged mACE2H353K mice, the mouse-adapted strain caused diffuse alveolar disease. The mouse-adapted virus also infected standard BALB/c and C57BL/6 mice, causing severe disease. The mouse-adapted virus acquired five new missense mutations including two in spike (K417E, Q493K), one each in nsp4, nsp9, and M and a single nucleotide change in the 5′ untranslated region. The Q493K spike mutation arose early in serial passage and is predicted to provide affinity-enhancing molecular interactions with mACE2 and further increase the stability and affinity to the receptor. This new model and mouse-adapted virus will be useful to evaluate COVID-19 disease and prophylactic and therapeutic interventions.","journal":"Journal of Virology","year":2024,"id":463394,"datarank":0.30008487024891417,"base_score":1.791759469228055,"endowment":1.791759469228055,"self_citation_contribution":0.26876392038420827,"citation_network_contribution":0.03132094986470591,"self_endowment_contribution":0.26876392038420827,"citer_contribution":0.03132094986470591,"corpus_percentile":null,"corpus_rank":null,"citation_count":5,"citer_count":4,"citers_with_citation_signal":2,"citers_with_endowment":2,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9585,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1293528,"name":"Abhishek Verma","orcid":"0000-0002-2374-5254","position":1,"is_corresponding":false},{"id":477099,"name":"Pengfei Li","orcid":"0000-0001-6436-4573","position":2,"is_corresponding":false},{"id":246347,"name":"Miguel Ortiz","orcid":"0000-0002-3573-5416","position":3,"is_corresponding":false},{"id":645808,"name":"Grant M. Hawkins","orcid":"0000-0003-0612-8948","position":4,"is_corresponding":false},{"id":398004,"name":"Nicholas Schnicker","orcid":"0000-0002-5189-4943","position":5,"is_corresponding":false},{"id":805262,"name":"Peter J. Szachowicz","orcid":"0000-0003-1735-297X","position":6,"is_corresponding":false},{"id":246349,"name":"Alejandro A. Pezzulo","orcid":"0000-0001-7544-5109","position":7,"is_corresponding":false},{"id":248538,"name":"Christine Wohlford-Lenane","orcid":null,"position":8,"is_corresponding":false},{"id":645806,"name":"Tom Kicmal","orcid":"0000-0001-7673-4690","position":9,"is_corresponding":false},{"id":227015,"name":"David K. Meyerholz","orcid":"0000-0003-1552-3253","position":10,"is_corresponding":false},{"id":241084,"name":"Tom Gallagher","orcid":"0000-0002-8601-5961","position":11,"is_corresponding":false},{"id":77538,"name":"Stanley Perlman","orcid":"0000-0003-4213-2354","position":12,"is_corresponding":false},{"id":225536,"name":"Paul B. McCray","orcid":"0000-0002-4067-577X","position":13,"is_corresponding":false},{"id":1053705,"name":"Kun Li","orcid":"0000-0002-8582-2913","position":0,"is_corresponding":true}],"reference_count":63,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:04:28.838123Z","pmid":"38168680","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}