{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,26]],"date-time":"2025-09-26T03:40:11Z","timestamp":1758858011715,"version":"3.44.0"},"reference-count":53,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2025,9,25]],"date-time":"2025-09-25T00:00:00Z","timestamp":1758758400000},"content-version":"vor","delay-in-days":24,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Gene Medicine"],"published-print":{"date-parts":[[2025,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Tertiary lymphoid structures (TLSs) are lymphoid structures found in non\u2010lymphoid tissues, which have been involved in the pathogenesis of multiple chronic diseases including tumors. TLSs are associated with better prognosis and response to immunotherapy in most types of tumors. However, what their associations with prognosis and the exact roles of TLS\u2010related genes in gallbladder cancer (GBC) are still unclear. Here, we investigated the associations of TLS with the prognosis in 85 patients with GBC. Our results showed that the presence of intra\u2010tumoral TLS was negatively associated with T stage (<jats:italic>p<\/jats:italic>\u2009=\u20090.035) and vascular invasion (<jats:italic>p<\/jats:italic>\u2009=\u20090.048) and predicted a higher rate of overall survival (<jats:italic>p<\/jats:italic>\u2009=\u20090.004) and a decreased risk of early recurrence (<jats:italic>p<\/jats:italic>\u2009=\u20090.002). Moreover, using integrative analysis of single\u2010cell RNA sequencing, bulk RNA sequencing, and deep machine learning, we screened six TLS\u2010related candidate genes in GBC, including CTSG, FLNC, CCNB1, HSPB8, NR4A1, and MYLK. Furthermore, through the evaluation of biofunctions and clinical significance, we found that CTSG, FLNC, CCNB1, and HSPB8 played an important role in immune infiltration, diagnosis, and prognosis of GBC. Finally, cell\u2010based experimental validation indicated that either knockdown of CTSG or overexpression of HSPB8 promoted CD8+ T cell apoptosis and exhaustion. In conclusion, TLS had a protective effect on human GBC, and its related genes, played a potential role in diagnosis, prognosis, immune infiltration, and metastasis of GBC. Our findings provided a new insight for the research on clinical biomarkers of GBC and development of its therapeutic targets.<\/jats:p>","DOI":"10.1002\/jgm.70037","type":"journal-article","created":{"date-parts":[[2025,9,25]],"date-time":"2025-09-25T23:00:36Z","timestamp":1758841236000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Multi\u2010Omics Integrated Analysis of the Protective Effect of Tertiary Lymphoid Structures and Associated Key Regulatory Genes in Human Gallbladder Cancer"],"prefix":"10.1002","volume":"27","author":[{"given":"Xiaolong","family":"Chen","sequence":"first","affiliation":[{"name":"National Key Clinical Department, Department of Hepatobiliary Surgery The First Affiliated Hospital of Chongqing Medical University  Chongqing China"},{"name":"Department of General Surgery, Xi'an No. 1 Hospital The First Affiliated Hospital of Northwestern University  Xi'an Shaanxi Province China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yimin","family":"Nong","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery Shenzhen Baoan Women's and Children's Hospital  Shenzhen Guangdong Province China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ming","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Hepatobiliary Pancreatic Surgery The First People's Hospital of Yibin  Yibin Sichuan Province China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chengyou","family":"Du","sequence":"additional","affiliation":[{"name":"National Key Clinical Department, Department of Hepatobiliary Surgery The First Affiliated Hospital of Chongqing Medical University  Chongqing China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,9,25]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41572-022-00398-y"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijsu.2022.106234"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.3748\/wjg.v30.i32.3739"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1097\/JS9.0000000000001403"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.abf9419"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2014.09.006"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41568-024-00728-0"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbcan.2025.189312"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-024-01992-w"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccell.2024.07.006"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-021-02356-5"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-023-06510-w"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijbiomac.2024.133918"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12967-024-05934-w"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2021.06.023"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2023-331773"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/advs.202406633"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41392-024-01947-5"},{"key":"e_1_2_11_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41423-020-0457-0"},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-23-3325"},{"key":"e_1_2_11_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2022.02.001"},{"key":"e_1_2_11_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccell.2024.09.007"},{"key":"e_1_2_11_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-1914-8"},{"key":"e_1_2_11_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-52153-4"},{"key":"e_1_2_11_26_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12072-024-10694-2"},{"key":"e_1_2_11_27_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-24-1173"},{"key":"e_1_2_11_28_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20231519"},{"key":"e_1_2_11_29_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-0979-8"},{"key":"e_1_2_11_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2020.108284"},{"key":"e_1_2_11_31_1","doi-asserted-by":"publisher","DOI":"10.1038\/s12276-025-01430-3"},{"key":"e_1_2_11_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2024.216693"},{"key":"e_1_2_11_33_1","doi-asserted-by":"publisher","DOI":"10.1097\/MD.0000000000036302"},{"key":"e_1_2_11_34_1","doi-asserted-by":"publisher","DOI":"10.1002\/1878-0261.13208"},{"key":"e_1_2_11_35_1","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2021.687915"},{"key":"e_1_2_11_36_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2025.1458341"},{"key":"e_1_2_11_37_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-021-00928-y"},{"key":"e_1_2_11_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2009.06.035"},{"key":"e_1_2_11_39_1","doi-asserted-by":"publisher","DOI":"10.2147\/OTT.S293148"},{"key":"e_1_2_11_40_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2011-07-368514"},{"key":"e_1_2_11_41_1","doi-asserted-by":"publisher","DOI":"10.1111\/ajco.13747"},{"key":"e_1_2_11_42_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12885-024-13412-1"},{"key":"e_1_2_11_43_1","doi-asserted-by":"publisher","DOI":"10.1177\/18758592251317400"},{"key":"e_1_2_11_44_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neurot.2025.e00569"},{"key":"e_1_2_11_45_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2018.11.019"},{"key":"e_1_2_11_46_1","doi-asserted-by":"publisher","DOI":"10.2174\/1871520614666140223191626"},{"key":"e_1_2_11_47_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsnano.1c04570"},{"key":"e_1_2_11_48_1","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2020.00545"},{"key":"e_1_2_11_49_1","doi-asserted-by":"publisher","DOI":"10.1002\/hep.31585"},{"key":"e_1_2_11_50_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells10020335"},{"key":"e_1_2_11_51_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejcb.2018.03.003"},{"key":"e_1_2_11_52_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12192-017-0780-2"},{"key":"e_1_2_11_53_1","doi-asserted-by":"publisher","DOI":"10.1155\/2012\/395329"},{"key":"e_1_2_11_54_1","doi-asserted-by":"publisher","DOI":"10.1038\/aps.2017.181"}],"container-title":["The Journal of Gene Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jgm.70037","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,9,26]],"date-time":"2025-09-26T03:26:51Z","timestamp":1758857211000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jgm.70037"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9]]},"references-count":53,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2025,9]]}},"alternative-id":["10.1002\/jgm.70037"],"URL":"https:\/\/doi.org\/10.1002\/jgm.70037","archive":["Portico"],"relation":{},"ISSN":["1099-498X","1521-2254"],"issn-type":[{"value":"1099-498X","type":"print"},{"value":"1521-2254","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9]]},"assertion":[{"value":"2025-04-30","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-08-30","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-09-25","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70037"}}