{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,14]],"date-time":"2026-02-14T15:12:45Z","timestamp":1771081965852,"version":"3.50.1"},"reference-count":8,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2026,12,12]],"date-time":"2026-12-12T00:00:00Z","timestamp":1797033600000},"content-version":"am","delay-in-days":365,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#am"},{"start":{"date-parts":[[2025,12,12]],"date-time":"2025-12-12T00:00:00Z","timestamp":1765497600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2025,12,12]],"date-time":"2025-12-12T00:00:00Z","timestamp":1765497600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000005","name":"U.S. Department of Defense","doi-asserted-by":"publisher","award":["CDMRP W81XWH\u201016\u20131\u20130544"],"award-info":[{"award-number":["CDMRP W81XWH\u201016\u20131\u20130544"]}],"id":[{"id":"10.13039\/100000005","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Prostate"],"published-print":{"date-parts":[[2026,3]]},"abstract":"ABSTRACT<\/jats:title>\n \n Background<\/jats:title>\n Molecular heterogeneity in prostate cancer is defined by distinct gene fusions and expression profiles involving ERG, SPINK1, ETV1, and ETV4. These markers are typically mutually exclusive and represent biologically distinct molecular subtypes with clinical and therapeutic relevance. Conventional methods assess these markers independently, limiting the ability to study their spatial relationships within the same tumor focus.<\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n We developed a combined dual immunohistochemistry (IHC) and dual RNA in situ hybridization (RNA\u2010ISH) assay for simultaneous detection of ERG, SPINK1, ETV1, and ETV4 in formalin\u2010fixed paraffin\u2010embedded (FFPE) prostate cancer tissues. Validated antibodies were used for ERG and SPINK1 protein detection, while RNAscope probes were employed for ETV1 and ETV4 mRNA visualization. The assay was optimized for sequential staining compatibility, chromogen contrast, and morphological preservation, and subsequently applied to prostate cancer cores representing various molecular subtypes.<\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n The combined assay enabled clear, concurrent visualization of ERG and SPINK1 protein expression together with ETV1 and ETV4 transcripts in a single tissue section. Expression patterns were mutually exclusive across tumor foci, consistent with known molecular subtypes of prostate cancer. The method maintained histological integrity and signal specificity, providing high\u2010resolution spatial information that could not be obtained by separate assays. This approach allowed detailed assessment of intra\u2010 and inter\u2010tumoral heterogeneity within the same histological context.<\/jats:p>\n <\/jats:sec>\n \n Conclusions<\/jats:title>\n This dual IHC and dual RNA\u2010ISH approach represents a novel and reliable platform for multiplex detection of key prostate cancer biomarkers on a single slide. The method offers significant advantages for molecular classification, tissue\u2010based biomarker validation, and comprehensive evaluation of tumor heterogeneity in translational and diagnostic research.<\/jats:p>\n <\/jats:sec>","DOI":"10.1002\/pros.70108","type":"journal-article","created":{"date-parts":[[2025,12,13]],"date-time":"2025-12-13T01:46:24Z","timestamp":1765590384000},"page":"493-504","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Mapping Molecular Diversity in Prostate Cancer With a Combined Multiplex IHC and RNA\u2010ISH Assay"],"prefix":"10.1002","volume":"86","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6950-0308","authenticated-orcid":false,"given":"Shannon","family":"Carskadon","sequence":"first","affiliation":[{"name":"Department of Urology Henry Ford Health Detroit MI USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3898-1460","authenticated-orcid":false,"given":"Sean","family":"Williamson","sequence":"additional","affiliation":[{"name":"Department of Genitourinary Pathology Cleveland Clinic OH USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-6533-4108","authenticated-orcid":false,"given":"Sangeetha","family":"Jyothilingam","sequence":"additional","affiliation":[{"name":"Department of Urology Henry Ford Health Detroit MI USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3740-5947","authenticated-orcid":false,"given":"Nilesh","family":"Gupta","sequence":"additional","affiliation":[{"name":"Department of Pathology Henry Ford Health Detroit MI USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0633-9772","authenticated-orcid":false,"given":"Nallasivam","family":"Palanisamy","sequence":"additional","affiliation":[{"name":"Department of Urology Henry Ford Health Detroit MI USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,12,12]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/modpathol.2012.234"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41379-020-0525-0"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/pros.23914"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.lfs.2023.122270"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.4103\/1008-682X.178852"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/labinvest.2010.179"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/modpathol.2015.148"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1097\/PAI.0000000000000095"}],"container-title":["The Prostate"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pros.70108","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/pros.70108","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/am-pdf\/10.1002\/pros.70108","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pros.70108","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,14]],"date-time":"2026-02-14T14:37:54Z","timestamp":1771079874000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/pros.70108"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,12]]},"references-count":8,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2026,3]]}},"alternative-id":["10.1002\/pros.70108"],"URL":"https:\/\/doi.org\/10.1002\/pros.70108","archive":["Portico"],"relation":{},"ISSN":["0270-4137","1097-0045"],"issn-type":[{"value":"0270-4137","type":"print"},{"value":"1097-0045","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,12]]},"assertion":[{"value":"2025-08-27","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-11-26","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-12","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}