{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T23:24:17Z","timestamp":1791415457131,"version":"4.3.3"},"reference-count":51,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T00:00:00Z","timestamp":1771286400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T00:00:00Z","timestamp":1771286400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100008545","name":"Georgia State University","doi-asserted-by":"publisher","award":["Research Initiation Grant (RIG)"],"award-info":[{"award-number":["Research Initiation Grant (RIG)"]}],"id":[{"id":"10.13039\/100008545","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Virol"],"published-print":{"date-parts":[[2026,2,17]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Venezuelan equine encephalitis virus (VEEV) is a prototypical encephalitic alphavirus. Members of the\n                      <jats:italic toggle=\"yes\">Alphavirus<\/jats:italic>\n                      genus are found across the globe, transmitted by arthropod vectors, and cause significant disease burdens in humans and animals. There are currently no FDA-approved antivirals for human use against any member of the\n                      <jats:italic toggle=\"yes\">Alphavirus<\/jats:italic>\n                      genus. While a vaccine exists against chikungunya virus (CHIKV), a member of the arthritogenic alphaviruses, FDA-approved vaccines are not available for other members of this genus, particularly the encephalitic alphaviruses such as VEEV, Eastern equine encephalitis virus, and Western equine encephalitis virus. 4\u2032-Fluorouridine (4\u2032-FlU, EIDD-2749) was recently identified as a broad-spectrum antiviral against multiple RNA viruses, including alphaviruses. 4\u2032-FlU can potently inhibit VEEV-TC83 replication, with submicromolar potency in cell culture. However, the emergence of antiviral resistance represents a hurdle for antiviral drug development and the implementation of effective treatment strategies. Here, we have identified novel mutations in the VEEV nsP4 RNA-dependent RNA polymerase that reduce susceptibility to 4\u2032-FlU, including P187A, Q191L, L289F, and T296I. We rebuilt each mutation in recombinant VEEV-TC83 and characterized the effects of these mutations on fitness and pathogenicity. In addition, we assessed the impact of mutations reducing sensitivity to 4\u2032-FlU in a mouse model. Although mutations against 4\u2032-FlU arise quickly\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      , treatment can still alleviate severe disease and lethal encephalitis. Together, these data highlight the promising therapeutic potential of 4\u2032-FlU for the treatment of alphavirus encephalitis.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Venezuelan equine encephalitis virus (VEEV) is a mosquito-spread virus that can cause encephalitis in people and animals. There are no FDA-approved countermeasures to treat VEEV infections in humans. 4\u2032-Fluorouridine (4\u2032-FlU) is currently being developed to treat multiple viral infections, including VEEV. A major problem with antivirals is the appearance of virus populations that are less susceptible to treatment. In this study, we treated infected mice with 4\u2032-FlU and measured how well the compound inhibited virus replication and prevented severe disease. In addition, we identified mutations in VEEV\u2019s polymerase that confer reduced susceptibility to 4\u2032-FlU. We then assessed if viruses encoding for these mutations were still pathogenic. Although VEEV can develop mild resistance to 4\u2032-FlU\n                        <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                        , administration of 4\u2032-FlU still reduced severe disease and prevented lethality in the animals infected with viruses that possess mutations that decrease susceptibility to 4\u2032-FlU. These results suggest that 4\u2032-FlU has strong potential as a future treatment for alphavirus infections.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jvi.01541-25","type":"journal-article","created":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T14:00:34Z","timestamp":1768485634000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":2,"title":["Impact of mutations affecting 4\u2032-fluorouridine susceptibility on fitness and treatment outcomes for Venezuelan equine encephalitis virus"],"prefix":"10.1128","volume":"100","author":[{"given":"Jenny","family":"Wong","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03qt6ba18","id-type":"ROR","asserted-by":"publisher"}],"name":"Center for Translational Antiviral Research, Georgia State University Institute for Biomedical Sciences","place":["Atlanta, 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