{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T20:10:46Z","timestamp":1782936646433,"version":"3.54.5"},"reference-count":25,"publisher":"American Society of Hematology","issue":"12","content-domain":{"domain":["ashpublications.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2026,6,23]]},"abstract":"<jats:sec>\n                    <jats:title>Abstract<\/jats:title>\n                    <jats:p>Diffuse large B-cell lymphoma (DLBCL) is a biologically heterogeneous disease. Two genomic classification systems, LymphGen and DLBclass, are capable of reproducibly classifying single tumors into subtypes with prognostic relevance in the context of standard-of-care R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) chemoimmunotherapy. Preliminary data from subgroup analyses of clinical trials suggest that the distinct biological features are targetable with drugs that exploit subtype-specific vulnerabilities, and large clinical trials are being designed to test this hypothesis. Although whole-exome sequencing (WES) remains the gold standard for each of these classification systems, smaller targeted sequencing panels reduce costs associated with sequencing, data storage, and processing. Here, we describe the design and validation of a targeted sequencing panel (LySeqST) that captures the genomic features required specifically for LymphGen classification. We perform in silico theoretical and real-world validation vs WES data and demonstrate that LySeqST can be used to accurately classify DLBCL tumors using both LymphGen and DLBclass. In an unselected population-based cohort, we determine the real-world proportions and validate the prognostic relevance of LymphGen subtypes, confirming that only tumors expressing the dark zone gene expression signature are consistently associated with inferior outcomes. Our results support the use of LySeqST for accurate genomic classification of DLBCL.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1182\/bloodadvances.2026020105","type":"journal-article","created":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T18:05:30Z","timestamp":1776276330000},"page":"4208-4214","update-policy":"https:\/\/doi.org\/10.1182\/blood.2019cm0000","source":"Crossref","is-referenced-by-count":1,"title":["LySeqST: a targeted sequencing assay for robust genomic classification of diffuse large B-cell lymphoma"],"prefix":"10.1182","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6413-6586","authenticated-orcid":false,"given":"Laura K.","family":"Hilton","sequence":"first","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"2Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3418-4958","authenticated-orcid":false,"given":"Sierra","family":"Gillis","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Faraneh","family":"Y-Moayyed","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5257-6277","authenticated-orcid":false,"given":"Joshua","family":"Bridgers","sequence":"additional","affiliation":[{"name":"3Department of Basic and Translational Research, BC Cancer Research Institute, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2365-5371","authenticated-orcid":false,"given":"Waleed","family":"Alduaij","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6153-7595","authenticated-orcid":false,"given":"Aixiang","family":"Jiang","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"4Department of Pathology and Laboratory Medicine, The University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2867-4037","authenticated-orcid":false,"given":"Susana","family":"Ben-Neriah","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5860-2798","authenticated-orcid":false,"given":"Brett","family":"Collinge","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8140-6517","authenticated-orcid":false,"given":"Jasper C. H.","family":"Wong","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7618-0103","authenticated-orcid":false,"given":"Merrill","family":"Boyle","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Meissner","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Graham W.","family":"Slack","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"4Department of Pathology and Laboratory Medicine, The University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9364-9391","authenticated-orcid":false,"given":"Pedro","family":"Farinha","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"4Department of Pathology and Laboratory Medicine, The University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1295-3258","authenticated-orcid":false,"given":"Jeffrey W.","family":"Craig","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"4Department of Pathology and Laboratory Medicine, The University of British Columbia, Vancouver, BC, Canada"},{"name":"5Department of Pathology, University of Virginia Health, Charlottesville, VA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4727-1425","authenticated-orcid":false,"given":"Alina S.","family":"Gerrie","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"6Division of Medical Oncology, Department of Medicine, The University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4625-3009","authenticated-orcid":false,"given":"Diego","family":"Villa","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"6Division of Medical Oncology, Department of Medicine, The University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5835-9863","authenticated-orcid":false,"given":"Kerry J.","family":"Savage","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"6Division of Medical Oncology, Department of Medicine, The University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1860-9765","authenticated-orcid":false,"given":"Laurie H.","family":"Sehn","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"6Division of Medical Oncology, Department of Medicine, The University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6753-892X","authenticated-orcid":false,"given":"Aly","family":"Karsan","sequence":"additional","affiliation":[{"name":"3Department of Basic and Translational Research, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"4Department of Pathology and Laboratory Medicine, The University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0003-0843","authenticated-orcid":false,"given":"George W.","family":"Wright","sequence":"additional","affiliation":[{"name":"7Biometric Research Branch, Division of Cancer Diagnosis and Treatment, National Cancer Institute, National Institutes of Health, Bethesda, MD"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1735-2892","authenticated-orcid":false,"given":"Louis M.","family":"Staudt","sequence":"additional","affiliation":[{"name":"8Lymphoid Malignancies Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2932-7800","authenticated-orcid":false,"given":"Ryan D.","family":"Morin","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"2Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada"},{"name":"3Department of Basic and Translational Research, BC Cancer Research Institute, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9842-9750","authenticated-orcid":false,"given":"Christian","family":"Steidl","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"4Department of Pathology and Laboratory Medicine, The University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0435-5947","authenticated-orcid":false,"given":"David W.","family":"Scott","sequence":"additional","affiliation":[{"name":"1Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada"},{"name":"6Division of Medical Oncology, Department of Medicine, The University of British Columbia, Vancouver, BC, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"234","published-online":{"date-parts":[[2026,6,11]]},"reference":[{"issue":"6769","key":"2026062308471700700_bib1","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1038\/35000501","article-title":"Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling","volume":"403","author":"Alizadeh","year":"2000","journal-title":"Nature"},{"issue":"3","key":"2026062308471700700_bib2","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1200\/JCO.18.01583","article-title":"Double-hit gene expression signature defines a distinct subgroup of germinal center B-cell-like diffuse large B-cell lymphoma","volume":"37","author":"Ennishi","year":"2019","journal-title":"J Clin Oncol"},{"issue":"20","key":"2026062308471700700_bib3","first-page":"2493","article-title":"Molecular determinants of clinical outcomes in a real-world diffuse large B-cell lymphoma population","volume":"141","author":"Alduaij","year":"2023","journal-title":"Blood"},{"issue":"28","key":"2026062308471700700_bib4","doi-asserted-by":"crossref","first-page":"4587","DOI":"10.1200\/JCO.2007.15.9277","article-title":"Addition of rituximab to standard chemotherapy improves the survival of both the germinal center B-cell\u2013like and non\u2013germinal center B-cell\u2013like subtypes of diffuse large B-cell lymphoma","volume":"26","author":"Fu","year":"2008","journal-title":"J Clin Oncol"},{"issue":"15","key":"2026062308471700700_bib5","doi-asserted-by":"crossref","first-page":"1396","DOI":"10.1056\/NEJMoa1801445","article-title":"Genetics and pathogenesis of diffuse large B-cell lymphoma","volume":"378","author":"Schmitz","year":"2018","journal-title":"N Engl J Med"},{"issue":"5","key":"2026062308471700700_bib6","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1038\/s41591-018-0016-8","article-title":"Molecular subtypes of diffuse large B cell lymphoma are associated with distinct pathogenic mechanisms and outcomes","volume":"24","author":"Chapuy","year":"2018","journal-title":"Nat Med"},{"issue":"4","key":"2026062308471700700_bib7","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1016\/j.ccell.2020.03.015","article-title":"A probabilistic classification tool for genetic subtypes of diffuse large B cell lymphoma with therapeutic implications","volume":"37","author":"Wright","year":"2020","journal-title":"Cancer Cell"},{"issue":"18","key":"2026062308471700700_bib8","doi-asserted-by":"crossref","first-page":"2041","DOI":"10.1182\/blood.2024025652","article-title":"DLBclass: a probabilistic molecular classifier to guide clinical investigation and practice in diffuse large B-cell lymphoma","volume":"145","author":"Chapuy","year":"2025","journal-title":"Blood"},{"issue":"20","key":"2026062308471700700_bib9","doi-asserted-by":"crossref","first-page":"1759","DOI":"10.1182\/blood.2019003535","article-title":"Targeted sequencing in DLBCL, molecular subtypes, and outcomes: a Haematological Malignancy Research Network report","volume":"135","author":"Lacy","year":"2020","journal-title":"Blood"},{"issue":"1","key":"2026062308471700700_bib10","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1038\/s41392-023-01358-y","article-title":"Simplified algorithm for genetic subtyping in diffuse large B-cell lymphoma","volume":"8","author":"Shen","year":"2023","journal-title":"Sig Transduct Target Ther"},{"issue":"4","key":"2026062308471700700_bib11","doi-asserted-by":"crossref","first-page":"814","DOI":"10.1111\/bjh.17811","article-title":"Molecular profiling in diffuse large B-cell lymphoma: why so many types of subtypes?","volume":"196","author":"Morin","year":"2022","journal-title":"Br J Haematol"},{"issue":"5","key":"2026062308471700700_bib12","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1053\/j.seminhematol.2023.11.006","article-title":"Biological heterogeneity in diffuse large B-cell lymphoma","volume":"60","author":"Hilton","year":"2023","journal-title":"Semin Hematol"},{"issue":"12","key":"2026062308471700700_bib13","doi-asserted-by":"crossref","first-page":"1643","DOI":"10.1016\/j.ccell.2021.10.006","article-title":"Effect of ibrutinib with R-CHOP chemotherapy in genetic subtypes of DLBCL","volume":"39","author":"Wilson","year":"2021","journal-title":"Cancer Cell"},{"issue":"10","key":"2026062308471700700_bib14","doi-asserted-by":"crossref","first-page":"1705","DOI":"10.1016\/j.ccell.2023.09.004","article-title":"Genetic subtype-guided immunochemotherapy in diffuse large B cell lymphoma: the randomized GUIDANCE-01 trial","volume":"41","author":"Zhang","year":"2023","journal-title":"Cancer Cell"},{"issue":"1","key":"2026062308471700700_bib15","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1093\/clinchem\/hvad147","article-title":"Chronic lymphocytic leukemia IGHV somatic hypermutation detection by targeted capture next-generation sequencing","volume":"70","author":"Grants","year":"2024","journal-title":"Clin Chem"},{"issue":"1","key":"2026062308471700700_bib16","doi-asserted-by":"crossref","first-page":"4001","DOI":"10.1038\/s41467-018-06354-3","article-title":"Genome-wide discovery of somatic regulatory variants in diffuse large B-cell lymphoma","volume":"9","author":"Arthur","year":"2018","journal-title":"Nat Commun"},{"issue":"25","key":"2026062308471700700_bib17","doi-asserted-by":"crossref","first-page":"4164","DOI":"10.1200\/JCO.23.00570","article-title":"Relapse timing is associated with distinct evolutionary dynamics in diffuse large B-cell lymphoma","volume":"41","author":"Hilton","year":"2023","journal-title":"J Clin Oncol"},{"issue":"7","key":"2026062308471700700_bib18","doi-asserted-by":"crossref","first-page":"2002","DOI":"10.1038\/s41375-021-01251-z","article-title":"Mutational mechanisms shaping the coding and noncoding genome of germinal center derived B-cell lymphomas","volume":"35","author":"H\u00fcbschmann","year":"2021","journal-title":"Leukemia"},{"issue":"24","key":"2026062308471700700_bib19","doi-asserted-by":"crossref","first-page":"2549","DOI":"10.1182\/blood.2022016095","article-title":"Minimal information for reporting a genomics experiment","volume":"140","author":"Dreval","year":"2022","journal-title":"Blood"},{"key":"2026062308471700700_bib20","doi-asserted-by":"publisher","DOI":"10.1101\/2025.09.19.676923","article-title":"LCR-modules: a collection of workflows for cancer genome analysis","author":"Dreval","year":"24  2025","journal-title":"bioRxiv"},{"issue":"8","key":"2026062308471700700_bib21","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.1182\/blood-2013-11-536433","article-title":"Determining cell-of-origin subtypes of diffuse large B-cell lymphoma using gene expression in formalin-fixed paraffin-embedded tissue","volume":"123","author":"Scott","year":"2014","journal-title":"Blood"},{"issue":"7","key":"2026062308471700700_bib22","first-page":"2326","article-title":"Validation of LymphGen classification on a 400-gene clinical next-generation sequencing panel in diffuse large B-cell lymphoma: real-world experience from a cancer center","volume":"109","author":"Zhu","year":"2024","journal-title":"Haematologica"},{"issue":"1","key":"2026062308471700700_bib23","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1111\/bjh.17132","article-title":"Application of the LymphGen classification tool to 928 clinically and genetically-characterised cases of diffuse large B cell lymphoma (DLBCL)","volume":"192","author":"Runge","year":"2021","journal-title":"Br J Haematol"},{"issue":"6","key":"2026062308471700700_bib24","doi-asserted-by":"crossref","first-page":"1929","DOI":"10.1182\/bloodadvances.2025017049","article-title":"Distribution and prognostic impact of LymphGen subtypes in Korean diffuse large B-cell lymphoma using clinical NGS panel","volume":"10","author":"Yim","year":"2026","journal-title":"Blood Adv"},{"key":"2026062308471700700_bib25","doi-asserted-by":"publisher","DOI":"10.64898\/2026.02.20.26346725","article-title":"LymphGen-Sig: integrating genetic and transcriptional states to predict therapeutic response in DLBCL","author":"Tumuluru","year":"2  2026","journal-title":"medRxiv"}],"container-title":["Blood Advances"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/ashpublications.org\/bloodadvances\/article-pdf\/10\/12\/4208\/2526841\/blooda_adv-2026-020105-main.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/ashpublications.org\/bloodadvances\/article-pdf\/10\/12\/4208\/2526841\/blooda_adv-2026-020105-main.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T12:50:42Z","timestamp":1782219042000},"score":1,"resource":{"primary":{"URL":"https:\/\/ashpublications.org\/bloodadvances\/article\/10\/12\/4208\/567780\/LySeqST-a-targeted-sequencing-assay-for-robust"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,11]]},"references-count":25,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2026,6,23]]}},"URL":"https:\/\/doi.org\/10.1182\/bloodadvances.2026020105","relation":{},"ISSN":["2473-9529","2473-9537"],"issn-type":[{"value":"2473-9529","type":"print"},{"value":"2473-9537","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2026,6,23]]},"published":{"date-parts":[[2026,6,11]]}}}