{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T17:10:25Z","timestamp":1785431425472,"version":"3.56.0"},"reference-count":101,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T00:00:00Z","timestamp":1782172800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T00:00:00Z","timestamp":1782172800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32372976"],"award-info":[{"award-number":["32372976"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Virol"],"published-print":{"date-parts":[[2026,6,23]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Long non-coding RNAs (LncRNAs) play pivotal regulatory roles in various biological processes, notably in immune regulation and viral infection. We previously identified the broad anti-influenza activity for LncRNA#61. Here, we further investigate the mechanism underlying its antiviral effect, both\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      . Using a lipid-nanoparticle-based delivery strategy, LncRNA#61 was successfully delivered into mice and effectively attenuated the replication and virulence of the highly pathogenic H5N1 influenza virus. Integrative transcriptomic analysis revealed that forced expression of LncRNA#61 markedly activated lipid metabolism, cell death, and Ragulator-Rag-mTORC1 pathways. Quantitative reverse transcription PCR analysis and a targeting metabolic assay further confirmed that LncRNA#61 is actively involved in regulating these pathways. Subsequent functional studies demonstrated that LncRNA#61 consistently enhances GSDMD-mediated pyroptosis both in murine LET-1 and canine MDCK cells. Notably, such pyroptosis was found to restrict H5N1 influenza virus replication. Intriguingly, ectopic expression of viral PA-X protein enhanced antiviral activity of LncRNA#61 both\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      . Mechanistically, PA-X interacts with LncRNA#61 and promotes LncRNA#61-mediated pyroptosis via a RagA-dependent reactive oxygen species pathway. Collectively, we here propose a novel model in which viral and host factors cooperate to activate a pro-death antiviral pathway. Our findings not only advance the fundamental knowledge of virus\u2013host interactions but also cross-link cell death, innate immunity, and metabolic regulation, pinpointing novel therapeutic targets against influenza.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      A current priority in anti-influenza research is developing broad-spectrum, host-directed therapeutics with low resistance risk. Here, we reveal that LncRNA#61-induced pyroptosis exerts an antiviral effect by restricting H5N1 virus replication both\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      , highlighting a novel cooperative virus\u2013host interaction that enhances antiviral immunity. Key contributions include the following: (i) identifying pyroptosis as a direct executioner mechanism that restricts H5N1 virus infection; (ii) revealing the unexpected role of forced expression of viral PA-X in augmenting antiviral activity of host LncRNA#61; and (iii) deciphering that LncRNA#61 interacts with PA-X and synergistically promotes GSDMD-mediated pyroptosis through a RagA\u2011ROS signaling cascade. Collectively, our work elucidates a novel antiviral mechanism wherein host LncRNA and viral protein co-opt the RagA-ROS-GSDMD axis to drive pyroptosis and inhibit viral replication. This discovery innovatively establishes a novel connection among viral pathogenesis, host cell death, and cellular metabolism, offering a fresh, integrative perspective for future studies on host-directed antiviral strategies.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/jvi.02214-25","type":"journal-article","created":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T13:17:15Z","timestamp":1778591835000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Long noncoding RNA#61 synergizes with viral PA-X to augment pyroptosis and attenuate the virulence of highly pathogenic H5N1 influenza virus in mice"],"prefix":"10.1128","volume":"100","author":[{"given":"Xia","family":"Chen","sequence":"first","affiliation":[{"name":"Key Laboratory of Avian Bioproducts Development, College of Veterinary Medicine, Yangzhou University","place":["Yangzhou, People's Republic of 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