{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T19:00:00Z","timestamp":1785438000875,"version":"3.56.0"},"reference-count":30,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T00:00:00Z","timestamp":1780012800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100009326","name":"Cancer Research Society","doi-asserted-by":"publisher","award":["1058389"],"award-info":[{"award-number":["1058389"]}],"id":[{"id":"10.13039\/100009326","id-type":"DOI","asserted-by":"publisher"}]},{"award":["1058389"],"award-info":[{"award-number":["1058389"]}],"id":[{"id":"https:\/\/ror.org\/00t38a349","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Michael Smith Health Research","award":["HPI-2024-03646"],"award-info":[{"award-number":["HPI-2024-03646"]}]},{"award":["HPI-2024-03646"],"award-info":[{"award-number":["HPI-2024-03646"]}],"id":[{"id":"https:\/\/ror.org\/020x39229","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background: Immunohistochemistry (IHC) plays a central role in subtyping of muscle-invasive bladder cancer (MIBC), yet conventional semi-quantitative scoring lacks objectivity and scalability. Automated digital pathology offers potential solutions, but requires robust, marker-specific validation against expert consensus scoring. Methods: We developed and internally validated an automated digital pathology pipeline for continuous IHC H-score quantification using QuPath (v0.6.0-arm64). Tissue microarrays (TMAs) were generated from transurethral resection of bladder tumor (TURBT) specimens from a cohort of patients with MIBC treated at Vancouver General Hospital. Cell detection was performed using StarDist (v0.9), followed by automated intensity-based H-score calculation for four epithelial IHC marker stains (CK14, CK20, CK5\/6, and Uroplakin II). H-scoring was then restricted to tumor epithelium by object-level classification using a supervised tumor\/non-tumor classifier trained on pathologist-reviewed annotations. Automated scores were compared with consensus scores from three blinded pathologists using Pearson correlation, linear regression, intraclass correlation coefficients (ICC), and Bland\u2013Altman analysis. Results: Automated H-scores demonstrated strong agreement with pathologist consensus across all four markers. CK14 showed near-perfect agreement (ICC \u2248 0.99) with minimal bias and narrow limits of agreement. CK20 also showed high agreement (ICC \u2248 0.95). CK5\/6 and Uroplakin II demonstrated slightly lower agreement (ICC \u2248 0.92 to 0.93) with mild proportional bias. Across markers, the automated pipeline preserved a broad H-score range, with range ratios of 0.96 to 0.99. Conclusions: This study establishes a robust, methods-forward pipeline for automated continuous IHC H-scoring in MIBC. The internally validated framework provides a scalable foundation for external cohort testing and future clinical outcome-associated biomarker analyses.<\/jats:p>","DOI":"10.3390\/diagnostics16111673","type":"journal-article","created":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T10:06:56Z","timestamp":1780049216000},"page":"1673","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Automated H-Scoring in Muscle-Invasive Bladder Cancer IHC: An Internal Validation Study"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-5247-246X","authenticated-orcid":false,"given":"Matthew","family":"Yap","sequence":"first","affiliation":[{"name":"Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 1Z7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0617-4178","authenticated-orcid":false,"given":"Ioana-Maria","family":"Mihai","sequence":"additional","affiliation":[{"name":"Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 1Z7, Canada"},{"name":"Pathology and Laboratory Medicine, BC Cancer, Vancouver, BC V5Z 4E6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maram Awadh A.","family":"Alanazi","sequence":"additional","affiliation":[{"name":"Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 1Z7, Canada"},{"name":"Anatomic Pathology, King Salman Specialist Hospital, Hail 55471, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5921-4634","authenticated-orcid":false,"given":"Gheorghe-Emilian","family":"Olteanu","sequence":"additional","affiliation":[{"name":"Pathology and Laboratory Medicine, BC Cancer, Vancouver, BC V5Z 4E6, Canada"},{"name":"Cellular Pathology Department, Royal Brompton Hospital, London SW3 6NP, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alberto","family":"Contreras-Sanz","sequence":"additional","affiliation":[{"name":"Department of Urologic Sciences, University of British Columbia, Vancouver, BC V6H 3Z6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2919-7068","authenticated-orcid":false,"given":"Peter","family":"Black","sequence":"additional","affiliation":[{"name":"Department of Urologic Sciences, University of British Columbia, Vancouver, BC V6H 3Z6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gang","family":"Wang","sequence":"additional","affiliation":[{"name":"Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 1Z7, Canada"},{"name":"Pathology and Laboratory Medicine, BC Cancer, Vancouver, BC V5Z 4E6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Yap, M., Mihai, I.M., and Wang, G. 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