{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T09:54:18Z","timestamp":1776938058138,"version":"3.51.4"},"reference-count":40,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T00:00:00Z","timestamp":1776902400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH\/NIDDK","award":["1R56DK129575"],"award-info":[{"award-number":["1R56DK129575"]}]},{"name":"startup funds"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Cancer immunotherapy has transformed oncology, including the management of urothelial carcinoma, yet response rates remain limited and mechanisms of resistance are incompletely understood. At the same time, recent initiatives from the National Institutes of Health and the United States Food and Drug Administration have emphasized the development of standardized human organoid platforms to improve preclinical modeling and reduce reliance on traditional animal systems. Urothelial cancer provides a compelling context in which to advance these efforts, given its established responsiveness to Bacillus Calmette\u2013Gu\u00e9rin and immune checkpoint blockade and its marked heterogeneity in clinical outcomes. Here, we review current organoid-based platforms for modeling bladder cancer immunotherapy, including patient biopsy-derived epithelial tumor organoids, air\u2013liquid interface cultures that retain endogenous stromal and immune components, engineered bladder cancer assembloids that reconstruct defined multicellular circuits, and induced pluripotent stem cell-derived urothelial organoids that offer renewable multilineage systems. For each approach, we outline strengths, technical constraints, and limitations in scalability, immune fidelity, and genetic matching. We conclude by discussing the critical challenges that must be addressed\u2014including benchmarking to patient tumors, reproducibility across laboratories, and standardized validation metrics\u2014to enable regulatory acceptance and clinical translation of organoid-based immunotherapy testing.<\/jats:p>","DOI":"10.3390\/cancers18091338","type":"journal-article","created":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T09:09:12Z","timestamp":1776935352000},"page":"1338","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Modeling Urothelial Carcinoma and Immunotherapy Response Using Organoid Platforms: From Epithelial Tumor Organoids to Multilineage Systems"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9435-275X","authenticated-orcid":false,"given":"Jorge O.","family":"M\u00fanera","sequence":"first","affiliation":[{"name":"Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC 29425, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e076743","DOI":"10.1136\/bmj-2023-076743","article-title":"Advances in diagnosis and treatment of bladder cancer","volume":"384","author":"Cookson","year":"2024","journal-title":"BMJ"},{"key":"ref_2","first-page":"10","article-title":"Cancer statistics, 2025","volume":"75","author":"Siegel","year":"2025","journal-title":"CA Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1016\/S0022-5347(17)58737-6","article-title":"Intracavitary bacillus calmette-guerin in the treatment of superficial bladder tumors","volume":"116","author":"Morales","year":"1976","journal-title":"J. Urol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1016\/S0022-5347(01)66424-3","article-title":"Urinary cytokines during intravesical bacillus calmette-guerin therapy for superficial bladder cancer: Processing, stability and prognostic value","volume":"155","author":"Kurth","year":"1996","journal-title":"J. Urol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1016\/S0022-5347(05)65469-9","article-title":"Prognostic value of a t helper 1 urinary cytokine response after intravesical bacillus calmette-guerin treatment for superficial bladder cancer","volume":"167","author":"Saint","year":"2002","journal-title":"J. Urol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"882","DOI":"10.1158\/1078-0432.CCR-19-1920","article-title":"Adaptive immune resistance to intravesical bcg in non-muscle invasive bladder cancer: Implications for prospective bcg-unresponsive trials","volume":"26","author":"Kates","year":"2020","journal-title":"Clin. Cancer Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1909","DOI":"10.1016\/S0140-6736(16)00561-4","article-title":"Atezolizumab in patients with locally advanced and metastatic urothelial carcinoma","volume":"387","author":"Rosenberg","year":"2016","journal-title":"Lancet"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1056\/NEJMoa1613683","article-title":"Pembrolizumab for advanced urothelial carcinoma after platinum-based chemotherapy","volume":"376","author":"Bellmunt","year":"2017","journal-title":"N. Engl. J. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1016\/S1470-2045(17)30065-7","article-title":"Nivolumab in metastatic urothelial carcinoma after platinum therapy (checkmate 275): A multicentre, single-arm, phase 2 trial","volume":"18","author":"Sharma","year":"2017","journal-title":"Lancet Oncol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e172411","DOI":"10.1001\/jamaoncol.2017.2411","article-title":"Efficacy and safety of durvalumab in locally advanced or metastatic urothelial carcinoma: Updated results from a phase 1\/2 open-label study","volume":"3","author":"Powles","year":"2017","journal-title":"JAMA Oncol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/S1470-2045(17)30900-2","article-title":"Avelumab in metastatic urothelial carcinoma after platinum failure (javelin solid tumor): Pooled results from two expansion cohorts of an open-label, phase 1 trial","volume":"19","author":"Patel","year":"2018","journal-title":"Lancet Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1218","DOI":"10.1056\/NEJMoa2002788","article-title":"Avelumab maintenance therapy for advanced or metastatic urothelial carcinoma","volume":"383","author":"Powles","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1483","DOI":"10.1016\/S1470-2045(17)30616-2","article-title":"First-line pembrolizumab in cisplatin-ineligible patients with locally advanced and unresectable or metastatic urothelial cancer (keynote-052): A multicentre, single-arm, phase 2 study","volume":"18","author":"Balar","year":"2017","journal-title":"Lancet Oncol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Snyder, A., Nathanson, T., Funt, S.A., Ahuja, A., Buros Novik, J., Hellmann, M.D., Chang, E., Aksoy, B.A., Al-Ahmadie, H., and Yusko, E. (2017). Contribution of systemic and somatic factors to clinical response and resistance to pd-l1 blockade in urothelial cancer: An exploratory multi-omic analysis. PLoS Med., 14.","DOI":"10.1371\/journal.pmed.1002309"},{"key":"ref_15","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":"Nature"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4287","DOI":"10.1158\/1078-0432.CCR-20-4574","article-title":"Myeloid cell-associated resistance to pd-1\/pd-l1 blockade in urothelial cancer revealed through bulk and single-cell rna sequencing","volume":"27","author":"Wang","year":"2021","journal-title":"Clin. Cancer Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1586","DOI":"10.1016\/j.cell.2016.05.082","article-title":"Modeling development and disease with organoids","volume":"165","author":"Clevers","year":"2016","journal-title":"Cell"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1016\/j.devcel.2016.08.014","article-title":"Organoids: Modeling development and the stem cell niche in a dish","volume":"38","author":"Kretzschmar","year":"2016","journal-title":"Dev. Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1146\/annurev-cellbio-100814-125218","article-title":"Mini-gut organoids: Reconstitution of the stem cell niche","volume":"31","author":"Date","year":"2015","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1016\/j.cell.2018.03.017","article-title":"Tumor evolution and drug response in patient-derived organoid models of bladder cancer","volume":"173","author":"Lee","year":"2018","journal-title":"Cell"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2214","DOI":"10.1038\/s41467-023-37696-2","article-title":"Bladder cancer organoids as a functional system to model different disease stages and therapy response","volume":"14","author":"Minoli","year":"2023","journal-title":"Nat. Commun."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4567","DOI":"10.1073\/pnas.1803595116","article-title":"Mouse and human urothelial cancer organoids: A tool for bladder cancer research","volume":"116","author":"Mullenders","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1038\/s41416-023-02494-6","article-title":"Urine-derived bladder cancer organoids (urinoids) as a tool for cancer longitudinal response monitoring and therapy adaptation","volume":"130","author":"Viergever","year":"2024","journal-title":"Br. J. Cancer"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e202317613","DOI":"10.1002\/anie.202317613","article-title":"Patient-derived immunocompetent tumor organoids: A platform for chemotherapy evaluation in the context of t-cell recognition","volume":"63","author":"Zhao","year":"2024","journal-title":"Angew. Chem. Int. Ed. Engl."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1972","DOI":"10.1016\/j.cell.2018.11.021","article-title":"Organoid modeling of the tumor immune microenvironment","volume":"175","author":"Neal","year":"2018","journal-title":"Cell"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1038\/nm.1951","article-title":"Sustained in vitro intestinal epithelial culture within a wnt-dependent stem cell niche","volume":"15","author":"Ootani","year":"2009","journal-title":"Nat. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"664","DOI":"10.1038\/s41586-020-3034-x","article-title":"Creation of bladder assembloids mimicking tissue regeneration and cancer","volume":"588","author":"Kim","year":"2020","journal-title":"Nature"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e1248","DOI":"10.1002\/cti2.1248","article-title":"Patient-derived organoids of bladder cancer recapitulate antigen expression profiles and serve as a personal evaluation model for car-t cells in vitro","volume":"10","author":"Yu","year":"2021","journal-title":"Clin. Transl. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"861","DOI":"10.1016\/j.cell.2007.11.019","article-title":"Induction of pluripotent stem cells from adult human fibroblasts by defined factors","volume":"131","author":"Takahashi","year":"2007","journal-title":"Cell"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1145","DOI":"10.1126\/science.282.5391.1145","article-title":"Embryonic stem cell lines derived from human blastocysts","volume":"282","author":"Thomson","year":"1998","journal-title":"Science"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"10506","DOI":"10.1038\/s41598-019-46848-8","article-title":"Directed differentiation of human induced pluripotent stem cells into mature stratified bladder urothelium","volume":"9","author":"Suzuki","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"7139","DOI":"10.3390\/ijms15057139","article-title":"Generation of bladder urothelium from human pluripotent stem cells under chemically defined serum- and feeder-free system","volume":"15","author":"Kang","year":"2014","journal-title":"Int. J. Mol. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1016\/j.eururo.2013.03.054","article-title":"A novel model of urinary tract differentiation, tissue regeneration, and disease: Reprogramming human prostate and bladder cells into induced pluripotent stem cells","volume":"64","author":"Moad","year":"2013","journal-title":"Eur. Urol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"610","DOI":"10.5966\/sctm.2013-0131","article-title":"Induction of human embryonic and induced pluripotent stem cells into urothelium","volume":"3","author":"Osborn","year":"2014","journal-title":"Stem Cells Transl. Med."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"112398","DOI":"10.1016\/j.isci.2025.112398","article-title":"Differentiation of human pluripotent stem cells into urothelial organoids via transient activation of wnt signaling","volume":"28","author":"Qu","year":"2025","journal-title":"iScience"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1201","DOI":"10.1038\/s41588-025-02182-6","article-title":"An integrated transcriptomic cell atlas of human endoderm-derived organoids","volume":"57","author":"Xu","year":"2025","journal-title":"Nat. Genet."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1021","DOI":"10.1038\/bjc.2014.166","article-title":"Guidelines for the use of cell lines in biomedical research","volume":"111","author":"Geraghty","year":"2014","journal-title":"Br. J. Cancer"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1434","DOI":"10.1016\/j.stem.2023.10.002","article-title":"Development of functional resident macrophages in human pluripotent stem cell-derived colonic organoids and human fetal colon","volume":"30","author":"Munera","year":"2023","journal-title":"Cell Stem Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.stem.2024.10.004","article-title":"Maturation and persistence of car t cells derived from human pluripotent stem cells via chemical inhibition of g9a\/glp","volume":"32","author":"Jing","year":"2025","journal-title":"Cell Stem Cell"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1181","DOI":"10.1016\/j.stem.2022.06.014","article-title":"Ezh1 repression generates mature ipsc-derived car t cells with enhanced antitumor activity","volume":"29","author":"Jing","year":"2022","journal-title":"Cell Stem Cell"}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/18\/9\/1338\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T09:17:50Z","timestamp":1776935870000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/18\/9\/1338"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,23]]},"references-count":40,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,5]]}},"alternative-id":["cancers18091338"],"URL":"https:\/\/doi.org\/10.3390\/cancers18091338","relation":{},"ISSN":["2072-6694"],"issn-type":[{"value":"2072-6694","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,23]]}}}