{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:18:45Z","timestamp":1785403125250,"version":"3.56.0"},"reference-count":32,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T00:00:00Z","timestamp":1775174400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100010211","name":"Education Department of Jilin Province","doi-asserted-by":"crossref","award":["JJKH20262142BS"],"award-info":[{"award-number":["JJKH20262142BS"]}],"id":[{"id":"10.13039\/501100010211","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100010211","name":"Education Department of Jilin Province","doi-asserted-by":"crossref","award":["JJKH20261232KJ"],"award-info":[{"award-number":["JJKH20261232KJ"]}],"id":[{"id":"10.13039\/501100010211","id-type":"DOI","asserted-by":"crossref"}]},{"award":["JJKH20262142BS"],"award-info":[{"award-number":["JJKH20262142BS"]}],"id":[{"id":"https:\/\/ror.org\/05782e512","id-type":"ROR","asserted-by":"publisher"}]},{"award":["JJKH20261232KJ"],"award-info":[{"award-number":["JJKH20261232KJ"]}],"id":[{"id":"https:\/\/ror.org\/05782e512","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Science and Technology Projects of Xizang Autonomous Region","award":["XZ202501ZY0143"],"award-info":[{"award-number":["XZ202501ZY0143"]}]},{"name":"Changsha Science and Technology Plan Natural Science Foundation","award":["kq2402188"],"award-info":[{"award-number":["kq2402188"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32330098"],"award-info":[{"award-number":["32330098"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"award":["32330098"],"award-info":[{"award-number":["32330098"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012226","name":"Fundamental Research Funds for the Central Universities","doi-asserted-by":"crossref","award":["SWU-KT22013"],"award-info":[{"award-number":["SWU-KT22013"]}],"id":[{"id":"10.13039\/501100012226","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Veterinary Sciences"],"abstract":"<jats:p>Background: The ORF3 protein of swine hepatitis E virus (HEV-4) is a key virulence factor involved in viral assembly, egress, and host signaling regulation. The mammalian target of rapamycin (mTOR) pathway plays a pivotal role in autophagy, metabolism, and immunity, and is often modulated by viruses to promote replication. However, it remains unknown whether HEV-4 ORF3 modulates the mTOR pathway via circular RNAs (circRNAs). Methods: Using an adenovirus-mediated ORF3 overexpression system in HepG2 cells, we integrated circRNA and transcriptome high-throughput sequencing data, followed by KEGG enrichment analysis to identify mTOR-associated differentially expressed genes. A circRNA\u2013miRNA regulatory network was constructed using bioinformatics tools, and the expression changes of m6A-related genes, including YTHDF3, were evaluated. Results: ORF3 overexpression significantly activated the mTOR pathway (p &lt; 0.05) and led to the identification of 20 mTOR-related circRNAs (e.g., circRNA5142). These circRNAs regulated downstream autophagy and lipid metabolism genes by sponging miRNAs such as hsa-let-7d-5p and hsa-miR-132-3p. Altered YTHDF3 expression indicated possible m6A-dependent epitranscriptomic regulation of the mTOR pathway. Conclusions: Our integrated analysis suggests that HEV-4 ORF3 may modulate the mTOR pathway through a circRNA\u2013miRNA network, perturbing host autophagy and metabolic balance, which may contribute to viral immune evasion. Targeting the ORF3-mediated circRNA-mTOR regulatory axis represents a promising therapeutic approach and provides a theoretical basis for novel anti-HEV-4 strategies.<\/jats:p>","DOI":"10.3390\/vetsci13040350","type":"journal-article","created":{"date-parts":[[2026,4,6]],"date-time":"2026-04-06T02:06:21Z","timestamp":1775441181000},"page":"350","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Integrated Analysis of the mTOR Signaling Pathway Mediated by the ORF3 Protein of Swine Hepatitis E Virus in HepG2 Cells via a circRNA\u2013miRNA Network"],"prefix":"10.3390","volume":"13","author":[{"given":"Jiya","family":"Li","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine, Jilin Agricultural University, Changchun 130118, China"},{"name":"The College of Veterinary Medicine, Southwest University, Chongqing 402460, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shengping","family":"Wu","sequence":"additional","affiliation":[{"name":"The College of Veterinary Medicine, Southwest University, Chongqing 402460, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingjie","family":"Wang","sequence":"additional","affiliation":[{"name":"The College of Veterinary Medicine, Southwest University, Chongqing 402460, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Cao","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Jilin Agricultural University, Changchun 130118, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yulong","family":"Yin","sequence":"additional","affiliation":[{"name":"Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leli","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanwei","family":"Jiao","sequence":"additional","affiliation":[{"name":"The College of Veterinary Medicine, Southwest University, Chongqing 402460, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"886","DOI":"10.1002\/vms3.707","article-title":"Research on a rat model of genotype IV swine hepatitis E virus","volume":"8","author":"Jian","year":"2022","journal-title":"Vet. 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