Establishment and characterization of primary retinoblastoma cell lines: a comprehensive molecular profiling approachDownload PDF Download PDF ArticleOpen accessPublished: 16 September 2026Angela Galardi ORCID: orcid.org/0000-0001-6945-49301 na1,Silvia Lampis1 na1,Virginia Di Paolo1,Lauriane Lemelle2,Valeria Vesprini1,Sara Patrizi ORCID: orcid.org/0000-0003-1898-15161,Luana Abballe ORCID: orcid.org/0000-0003-4446-22931,Valentina Di Ruscio ORCID: orcid.org/0000-0002-9581-27511,Sabina Barresi3,Isabella Giovannoni ORCID: orcid.org/0000-0002-9980-65943,Gemma D’Elia4,Emanuele Agolini4,Rita De Vito3,Paola Valente5,Antonino Romanzo5,Angela Mastronuzzi1,Debora De Pasquale1,Rita Alaggio3,Franco Locatelli1,6,Ida Russo1 na2,Evelina Miele ORCID: orcid.org/0000-0002-4747-10321 na2 &…Angela Di Giannatale1 na2 npj Precision Oncology (2026) Cite this articleSave articleView saved research We’re sharing this article early to provide faster access to peer-reviewed, accepted research. It is citable and carries a permanent DOI. This version is subject to further edits and will be replaced automatically by the final Version of Record. All legal disclaimers apply.AbstractRetinoblastoma (RB) research is significantly constrained by the limited availability of preclinical models that accurately recapitulate the disease complexity, particularly regarding the context of vitreous seeding, a major driver of therapeutic resistance and treatment failure. Here, we report the establishment and integrative characterization of patient-derived, three-dimensional (3D) spheroid models generated from primary retinal tumors (RBT) and vitreous seeds (RBVS). Across a cohort of 10 cell lines generated from 8 eyes and 8 patients, both RBT and RBVS models demonstrated sustained 3D growth and preserved key phenotypic features of the original tumors. Genome-wide DNA methylation and copy number profiling revealed strong concordance between primary tumors and derived models, supporting their molecular fidelity. RB1 inactivation was identified as the founding driver event, while additional alterations, including BCOR loss-of-function mutations and MYCN, ALK, and IKBKE amplifications, defined clinically aggressive subsets and were retained in corresponding models. Comparative analyses of matched samples showed that 3D spheroid generated models maintain core oncogenic drivers. Collectively, our data establish patient-derived 3D RB models as robust and translationally relevant platforms that recapitulate tumor heterogeneity and evolution. These systems provide a valuable resource for biomarker discovery, mechanistic investigation of resistance pathways, and development of precision therapeutic strategies in RB.AcknowledgementsThis work was supported by the Italian Ministry of Health with RC 2026. We would like to thank Fondation Nuovo Soldati and Institut Servier for their support of Lauriane Lemelle.Author informationAuthor notesThese authors contributed equally: Angela Galardi, Silvia Lampis.These authors jointly supervised this work: Ida Russo, Evelina Miele, Angela Di Giannatale.Authors and AffiliationsDepartment of Oncology/Hematology, Gene and Cell Therapy and Hemopoietic Transplant, Bambino Gesù Children’s Hospital, IRCCS, Rome, ItalyAngela Galardi, Silvia Lampis, Virginia Di Paolo, Valeria Vesprini, Sara Patrizi, Luana Abballe, Valentina Di Ruscio, Angela Mastronuzzi, Debora De Pasquale, Franco Locatelli, Ida Russo, Evelina Miele & Angela Di GiannataleSIREDO Oncology Center (Care, Innovation and Research for Children and AYA with Cancer), PSL Research University, Institut Curie, Paris, FranceLauriane LemellePathology Unit, Department of Laboratories, Bambino Gesù Children’s Hospital, IRCCS, Rome, ItalySabina Barresi, Isabella Giovannoni, Rita De Vito & Rita AlaggioLaboratory of Medical Genetics, Bambino Gesù Children’s Hospital, IRCCS, Rome, ItalyGemma D’Elia & Emanuele AgoliniOphthalmology Unit, Bambino Gesù Children’s Hospital, IRCCS, Rome, ItalyPaola Valente & Antonino RomanzoDepartment of Life Sciences and Public Health, Catholic University of the Sacred Heart, Rome, ItalyFranco LocatelliAuthorsAngela GalardiView author publicationsSearch author on:PubMed Google ScholarSilvia LampisView author publicationsSearch author on:PubMed Google ScholarVirginia Di PaoloView author publicationsSearch author on:PubMed Google ScholarLauriane LemelleView author publicationsSearch author on:PubMed Google ScholarValeria VespriniView author publicationsSearch author on:PubMed Google ScholarSara PatriziView author publicationsSearch author on:PubMed Google ScholarLuana AbballeView author publicationsSearch author on:PubMed Google ScholarValentina Di RuscioView author publicationsSearch author on:PubMed Google ScholarSabina BarresiView author publicationsSearch author on:PubMed Google ScholarIsabella GiovannoniView author publicationsSearch author on:PubMed Google ScholarGemma D’EliaView author publicationsSearch author on:PubMed Google ScholarEmanuele AgoliniView author publicationsSearch author on:PubMed Google ScholarRita De VitoView author publicationsSearch author on:PubMed Google ScholarPaola ValenteView author publicationsSearch author on:PubMed Google ScholarAntonino RomanzoView author publicationsSearch author on:PubMed Google ScholarAngela MastronuzziView author publicationsSearch author on:PubMed Google ScholarDebora De PasqualeView author publicationsSearch author on:PubMed Google ScholarRita AlaggioView author publicationsSearch author on:PubMed Google ScholarFranco LocatelliView author publicationsSearch author on:PubMed Google ScholarIda RussoView author publicationsSearch author on:PubMed Google ScholarEvelina MieleView author publicationsSearch author on:PubMed Google ScholarAngela Di GiannataleView author publicationsSearch author on:PubMed Google ScholarCorresponding authorCorrespondence to Virginia Di Paolo.Ethics declarationsCompeting interestsThe authors declare no competing interests.Additional informationPublisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.Supplementary informationRights and permissionsOpen Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. 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