{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T00:15:26Z","timestamp":1790727326611,"version":"4.1.0"},"reference-count":46,"publisher":"BMJ","issue":"7","license":[{"start":{"date-parts":[[2024,7,23]],"date-time":"2024-07-23T00:00:00Z","timestamp":1721692800000},"content-version":"unspecified","delay-in-days":22,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["20K07601; 23K06778"],"award-info":[{"award-number":["20K07601; 23K06778"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Immunother Cancer"],"accepted":{"date-parts":[[2024,6,27]]},"published-print":{"date-parts":[[2024,7]]},"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Anti-programmed death-1 (PD-1)\/anti-PD-ligand-1 (PD-L1) pathway inhibition is a standard regimen for advanced urothelial carcinoma (UC); however, its limited efficacy has been reflected in reported medium response rates. This study explored the role of next-generation coinhibitory receptors (IRs; lymphocyte activation gene 3 (LAG-3), T-cell immunoglobulin and mucin domain 3 (TIM-3), and T-cell immunoreceptor with Ig and ITIM domains (TIGIT)) and their ligands (LGs) in the response to PD-(L)1 blockade therapy and the oncological outcomes in patients with UC.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We investigated metastatic UC cases who underwent PD-(L)1 therapy (cohort 1: n=348, cohort 2: n=89, and cohort 4: n=29) or advanced UC cases involving surgery (cohort 3: n=293\u2009and cohort 5: n=90). We assessed the mRNA expression profiles and corresponding clinical information regarding IRs and LGs using cohorts 1, 2, and 3. Additionally, we elucidated the spatial features of these targeted markers using multiplex immunohistochemistry (mIHC) on formalin-fixed paraffin-embedded samples from cohorts 4 and 5. Survival, differential expressed gene, and Gene Set Enrichment analyses were performed. For mIHC, quantitative analyses were also performed to correlate immune and tumor cell densities with patient survival.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      <jats:italic>LAG-3<\/jats:italic>\n                      expression was strongly associated with the responsiveness of PD-(L)1 blockade compared with the expression of\n                      <jats:italic>TIM-3<\/jats:italic>\n                      and\n                      <jats:italic>TIGIT<\/jats:italic>\n                      . In tumors with high\n                      <jats:italic>LAG-3<\/jats:italic>\n                      levels, the increased expression of\n                      <jats:italic>fibrinogen-like protein 1<\/jats:italic>\n                      (\n                      <jats:italic>FGL1<\/jats:italic>\n                      ) had a significantly negative effect on the response to PD-(L)1 blockade and overall survival. Moreover, high\n                      <jats:italic>FGL1<\/jats:italic>\n                      levels were associated with elevated CD4\n                      <jats:sup>+<\/jats:sup>\n                      regulatory T-cell gene signatures and the upregulation of\n                      <jats:italic>CD39<\/jats:italic>\n                      and\n                      <jats:italic>neuropilin-1<\/jats:italic>\n                      , with both indicating CD8\n                      <jats:sup>+<\/jats:sup>\n                      T-cell exhaustion. mIHC analyses revealed that patients with stromal CD8\n                      <jats:sup>+<\/jats:sup>\n                      LAG-3\n                      <jats:sup>+<\/jats:sup>\n                      cells\n                      <jats:sup>high<\/jats:sup>\n                      \u2013tumor FGL1\n                      <jats:sup>+<\/jats:sup>\n                      cells\n                      <jats:sup>high<\/jats:sup>\n                      exhibited a significant negative correlation with survival rates compared with those with stromal CD8\n                      <jats:sup>+<\/jats:sup>\n                      LAG-3\n                      <jats:sup>+<\/jats:sup>\n                      cells\n                      <jats:sup>high<\/jats:sup>\n                      \u2013tumor FGL1\n                      <jats:sup>+<\/jats:sup>\n                      cells\n                      <jats:sup>low<\/jats:sup>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>LAG-3 expression and high FGL1 coexpression are important predictive factors of adverse oncological outcomes related to the presence of immunosuppressive contextures. These findings are hypothesis-generating, warranting further mechanistic and clinical studies aimed to evaluate LAG-3\/FGL1 blockade in UC.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1136\/jitc-2024-009358","type":"journal-article","created":{"date-parts":[[2024,7,23]],"date-time":"2024-07-23T22:37:15Z","timestamp":1721774235000},"page":"e009358","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":17,"title":["Impact of LAG-3\/FGL1 pathway on immune evasive contexture and clinical outcomes in advanced urothelial carcinoma"],"prefix":"10.1136","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1119-312X","authenticated-orcid":false,"given":"Takashi","family":"Yoshida","sequence":"first","affiliation":[{"name":"Department of Urology and Andrology, Kansai Medical University, Osaka, Japan"},{"name":"Graduate School of Engineering, Tottori University, Tottori, Japan"},{"name":"Department of Urology, Osaka Saiseikai-Noe Hospital, Osaka, Japan"},{"name":"Corporate Sponsored Research Programs for Multicellular Interactions in Cancer, Kansai Medical University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-7540-1812","authenticated-orcid":false,"given":"Takahiro","family":"Nakamoto","sequence":"additional","affiliation":[{"name":"Department of Urology and Andrology, Kansai Medical University, Osaka, Japan"},{"name":"Department of Pathology, Kansai Medical University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naho","family":"Atsumi","sequence":"additional","affiliation":[{"name":"Corporate Sponsored Research Programs for Multicellular Interactions in Cancer, Kansai Medical University, Osaka, Japan"},{"name":"Department of Pathology, Kansai Medical University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chisato","family":"Ohe","sequence":"additional","affiliation":[{"name":"Department of Urology and Andrology, Kansai Medical University, Osaka, Japan"},{"name":"Department of Pathology, Graduate School of Medicine, Osaka Metropolitan University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takeshi","family":"Sano","sequence":"additional","affiliation":[{"name":"Department of Urology and Andrology, Kansai Medical University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshiki","family":"Yasukochi","sequence":"additional","affiliation":[{"name":"Department of Genome Analysis, Institute of Biomedical Science, Kansai Medical University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Koji","family":"Tsuta","sequence":"additional","affiliation":[{"name":"Corporate Sponsored Research Programs for Multicellular Interactions in Cancer, Kansai Medical University, Osaka, Japan"},{"name":"Department of Pathology, Kansai Medical University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hidefumi","family":"Kinoshita","sequence":"additional","affiliation":[{"name":"Department of Urology and Andrology, Kansai Medical University, Osaka, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"239","published-online":{"date-parts":[[2024,7,23]]},"reference":[{"key":"2025100315125900000_12.7.e009358.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.eururo.2020.03.055"},{"key":"2025100315125900000_12.7.e009358.2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2034442"},{"key":"2025100315125900000_12.7.e009358.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccell.2020.03.017"},{"key":"2025100315125900000_12.7.e009358.4","doi-asserted-by":"crossref","first-page":"1608","DOI":"10.1200\/JCO.19.00538","article-title":"Nivolumab alone and with Ipilimumab in previously treated metastatic urothelial carcinoma: checkmate 032 Nivolumab 1 mg\/kg plus Ipilimumab 3 mg\/kg expansion cohort results","volume":"37","author":"Sharma","year":"2019","journal-title":"JCO"},{"key":"2025100315125900000_12.7.e009358.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejca.2021.03.049"},{"key":"2025100315125900000_12.7.e009358.6","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2016.05.001"},{"key":"2025100315125900000_12.7.e009358.7","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-019-0512-0"},{"key":"2025100315125900000_12.7.e009358.8","doi-asserted-by":"publisher","DOI":"10.1016\/S1470-2045(22)00226-1"},{"key":"2025100315125900000_12.7.e009358.9","doi-asserted-by":"publisher","DOI":"10.1158\/1078-0432.CCR-20-4746"},{"key":"2025100315125900000_12.7.e009358.10","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2109970"},{"key":"2025100315125900000_12.7.e009358.11","doi-asserted-by":"crossref","DOI":"10.1080\/2162402X.2023.2261248","article-title":"Lag-3 expression and clinical outcomes in metastatic melanoma patients treated with combination anti-lag-3 + anti-PD-1-based immunotherapies","volume":"12","author":"Gide","year":"2023","journal-title":"Oncoimmunology"},{"key":"2025100315125900000_12.7.e009358.12","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(16)32455-2"},{"key":"2025100315125900000_12.7.e009358.13","doi-asserted-by":"crossref","first-page":"544","DOI":"10.1038\/nature25501","article-title":"TGFbeta attenuates tumour response to PD-L1 blockade by contributing to exclusion of T cells","volume":"554","author":"Mariathasan","year":"2018","journal-title":"Nat New Biol"},{"key":"2025100315125900000_12.7.e009358.14","doi-asserted-by":"publisher","DOI":"10.1038\/s41416-021-01488-6"},{"key":"2025100315125900000_12.7.e009358.15","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.25514"},{"key":"2025100315125900000_12.7.e009358.16","doi-asserted-by":"crossref","DOI":"10.1038\/s41467-021-22625-y","article-title":"A clinical transcriptome approach to patient stratification and therapy selection in acute myeloid leukemia","volume":"12","author":"Docking","year":"2021","journal-title":"Nat Commun"},{"key":"2025100315125900000_12.7.e009358.17","doi-asserted-by":"publisher","DOI":"10.1182\/bloodadvances.2016000786"},{"key":"2025100315125900000_12.7.e009358.18","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0506580102"},{"key":"2025100315125900000_12.7.e009358.19","doi-asserted-by":"publisher","DOI":"10.1038\/ng1180"},{"key":"2025100315125900000_12.7.e009358.20","doi-asserted-by":"publisher","DOI":"10.1101\/gr.239244.118"},{"key":"2025100315125900000_12.7.e009358.21","doi-asserted-by":"crossref","DOI":"10.1136\/jitc-2022-004569","article-title":"NKG2A and PD-L1 expression panel predicts clinical benefits from adjuvant chemotherapy and PD-L1 blockade in muscle-invasive bladder cancer","volume":"10","author":"Yan","year":"2022","journal-title":"J Immunother Cancer"},{"key":"2025100315125900000_12.7.e009358.22","doi-asserted-by":"crossref","DOI":"10.1136\/jitc-2020-002197","article-title":"Multi-institutional TSA-amplified multiplexed immunofluorescence reproducibility evaluation (MITRE) study","volume":"9","author":"Taube","year":"2021","journal-title":"J Immunother Cancer"},{"key":"2025100315125900000_12.7.e009358.23","doi-asserted-by":"crossref","first-page":"606","DOI":"10.1038\/s41416-021-01628-y","article-title":"Pretreatment tumour immune microenvironment predicts clinical response and prognosis of muscle-invasive bladder cancer in the neoadjuvant chemotherapy setting","volume":"126","author":"Ikarashi","year":"2022","journal-title":"Br J Cancer"},{"key":"2025100315125900000_12.7.e009358.24","doi-asserted-by":"crossref","DOI":"10.1038\/s41598-017-13942-8","article-title":"Validation of multiplex immunofluorescence panels using multispectral microscopy for immune-profiling of formalin-fixed and paraffin-embedded human tumor tissues","volume":"7","author":"Parra","year":"2017","journal-title":"Sci Rep"},{"key":"2025100315125900000_12.7.e009358.25","doi-asserted-by":"crossref","DOI":"10.1038\/s41467-019-11788-4","article-title":"Macrophage spatial heterogeneity in gastric cancer defined by multiplex immunohistochemistry","volume":"10","author":"Huang","year":"2019","journal-title":"Nat Commun"},{"key":"2025100315125900000_12.7.e009358.26","doi-asserted-by":"crossref","first-page":"3001","DOI":"10.1007\/s00262-021-03006-2","article-title":"Multiplexed single-cell pathology reveals the association of CD8 T-cell heterogeneity with prognostic outcomes in renal cell carcinoma","volume":"70","author":"Murakami","year":"2021","journal-title":"Cancer Immunol Immunother"},{"key":"2025100315125900000_12.7.e009358.27","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2018.11.010"},{"key":"2025100315125900000_12.7.e009358.28","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2022.03.013"},{"key":"2025100315125900000_12.7.e009358.29","doi-asserted-by":"publisher","DOI":"10.1038\/nri1708"},{"key":"2025100315125900000_12.7.e009358.30","doi-asserted-by":"publisher","DOI":"10.1158\/2326-6066.CIR-14-0150"},{"key":"2025100315125900000_12.7.e009358.31","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0903474106"},{"key":"2025100315125900000_12.7.e009358.32","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1038\/nature13848","article-title":"CEACAM1 regulates TIM-3-mediated tolerance and exhaustion","volume":"517","author":"Huang","year":"2015","journal-title":"Nat New Biol"},{"key":"2025100315125900000_12.7.e009358.33","doi-asserted-by":"publisher","DOI":"10.1038\/ni.2376"},{"key":"2025100315125900000_12.7.e009358.34","doi-asserted-by":"publisher","DOI":"10.1038\/ni1271"},{"key":"2025100315125900000_12.7.e009358.35","doi-asserted-by":"publisher","DOI":"10.1016\/j.juro.2015.08.082"},{"key":"2025100315125900000_12.7.e009358.36","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1817323"},{"key":"2025100315125900000_12.7.e009358.37","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1038\/nature12428","article-title":"Stability and function of regulatory T cells is maintained by a Neuropilin-1-semaphorin-4A axis","volume":"501","author":"Delgoffe","year":"2013","journal-title":"Nature New Biol"},{"key":"2025100315125900000_12.7.e009358.38","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.05.005"},{"key":"2025100315125900000_12.7.e009358.39","doi-asserted-by":"publisher","DOI":"10.1186\/s13045-021-01161-8"},{"key":"2025100315125900000_12.7.e009358.40","doi-asserted-by":"crossref","DOI":"10.1136\/jitc-2021-004440","article-title":"Role of tumor infiltrating lymphocytes and spatial immune heterogeneity in sensitivity to PD-1 axis blockers in non-small cell lung cancer","volume":"10","author":"Lopez de Rodas","year":"2022","journal-title":"J Immunother Cancer"},{"key":"2025100315125900000_12.7.e009358.41","doi-asserted-by":"crossref","DOI":"10.1038\/s41389-020-00283-x","article-title":"Nuclear FOXP3 inhibits tumor growth and induced apoptosis in hepatocellular carcinoma by targeting c-Myc","volume":"9","author":"Gong","year":"2020","journal-title":"Oncogenesis"},{"key":"2025100315125900000_12.7.e009358.42","doi-asserted-by":"crossref","first-page":"13155","DOI":"10.1002\/cam4.6000","article-title":"LAG-3 transcriptomic expression patterns across malignancies: implications for precision immunotherapeutics","volume":"12","author":"Adashek","year":"2023","journal-title":"Cancer Med"},{"key":"2025100315125900000_12.7.e009358.43","doi-asserted-by":"crossref","first-page":"368","DOI":"10.62347\/MVDH5025","article-title":"LAG-3 transcriptomic expression correlates linearly with other checkpoints, but not with clinical outcomes","volume":"14","author":"Adashek","year":"2024","journal-title":"Am J Cancer Res"},{"key":"2025100315125900000_12.7.e009358.44","doi-asserted-by":"publisher","DOI":"10.1038\/nri3790"},{"key":"2025100315125900000_12.7.e009358.45","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1613683"},{"key":"2025100315125900000_12.7.e009358.46","doi-asserted-by":"crossref","DOI":"10.1136\/jitc-2020-000967","article-title":"Neuropilin-1: a checkpoint target with unique implications for cancer immunology and immunotherapy","volume":"8","author":"Chuckran","year":"2020","journal-title":"J Immunother Cancer"}],"container-title":["Journal for ImmunoTherapy of Cancer"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1136\/jitc-2024-009358","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,3]],"date-time":"2025-10-03T22:13:05Z","timestamp":1759529585000},"score":1,"resource":{"primary":{"URL":"https:\/\/jitc.bmj.com\/lookup\/doi\/10.1136\/jitc-2024-009358"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,7]]},"references-count":46,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2024,7,23]]},"published-print":{"date-parts":[[2024,7]]}},"alternative-id":["10.1136\/jitc-2024-009358"],"URL":"https:\/\/doi.org\/10.1136\/jitc-2024-009358","relation":{},"ISSN":["2051-1426"],"issn-type":[{"value":"2051-1426","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,7]]}}}