{"doi":"10.5281/zenodo.7307413","title":"Human Norovirus Efficiently Replicates in Differentiated 3D-Human Intestinal Enteroids","abstract":"Human norovirus (HNoV) accounts for one-fifth of all acute viral gastroenteri- tis worldwide and an economic burden of ;$60 billion globally. The lack of treatment options against HNoV is in part due to the lack of cultivation systems. Recently, a model of infection in biopsy-derived human intestinal enteroids (HIE) has been described: 3D-HIE are first dispersed in 2D-monolayers and differentiated prior to infection, resulting in a labor-intensive, time-consuming procedure. Here, we present an alternative protocol for HNoV infection of 3D-HIE. We found that 3D-HIE differentiated as efficiently as 2D-mono- layers. In addition, immunofluorescence-based quantification of UEA-1, a lectin that stains the villus brush border, revealed that ;80% of differentiated 3D-HIE spontaneously undergo polarity inversion, allowing for viral infection without the need for microinjection. Infection with HNoV GII.4-positive stool samples attained a fold-increase over inoculum of ;2 Log10 at 2 days postinfection or up to 3.5 Log10 when ruxolitinib, a JAK1/2-inhibitor, was added. Treatment of GII.4-infected 3D-HIE with the polymerase inhibitor 29-C- Methylcytidine (2CMC) and other antivirals showed a reduction in viral infection, suggest- ing that 3D-HIE are an excellent platform to test anti-infectives. The transcriptional host response to HNoV was then investigated by RNA sequencing in infected versus uninfected 3D-HIE in the presence of ruxolitinib to focus on virus-associated signatures while limiting interferon-stimulated gene signatures. The analysis revealed upregulated hormone and neurotransmitter signal transduction pathways and downregulated glycolysis and hypoxia- response pathways upon HNoV infection. Overall, 3D-HIE have proven to be a highly ro- bust model to study HNoV infection, screen antivirals, and to investigate the host response to HNoV infection.","journal":"Zenodo (CERN European Organization for Nuclear Research)","year":2022,"id":312963,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9532,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":868938,"name":"Nânci Santos-Ferreira","orcid":"0000-0003-4995-5580","position":1,"is_corresponding":false},{"id":109658,"name":"Merritt Gillilland","orcid":"0000-0002-6750-8541","position":2,"is_corresponding":false},{"id":583279,"name":"Roberto J. Cieza","orcid":"0000-0001-7400-722X","position":3,"is_corresponding":false},{"id":324124,"name":"Justin A. Colacino","orcid":"0000-0002-5882-4569","position":4,"is_corresponding":false},{"id":288649,"name":"Jonathan Z. Sexton","orcid":"0000-0002-9244-5888","position":5,"is_corresponding":false},{"id":218441,"name":"Johan Neyts","orcid":"0000-0002-0033-7514","position":6,"is_corresponding":false},{"id":715818,"name":"Stefan Taube","orcid":"0000-0002-9159-3651","position":7,"is_corresponding":false},{"id":218431,"name":"Joana Rocha‐Pereira","orcid":"0000-0001-8299-3787","position":8,"is_corresponding":false},{"id":288650,"name":"Christiane E. Wobus","orcid":"0000-0001-5286-0924","position":9,"is_corresponding":false},{"id":93854,"name":"Carmen Mirabelli","orcid":"0000-0002-4785-5482","position":0,"is_corresponding":true}],"reference_count":0,"raw_metadata":null,"created_at":"2026-07-19T00:33:40.460845Z","pmid":null,"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":[]}