De novo H3.3K27M-altered diffuse midline glioma in human brainstem organoids to dissect GD2 CAR T cell function
Diffuse midline glioma (DMG) is a highly aggressive and untreatable pediatric cancer primarily arising in the pontine brainstem region, necessitating the development of representative models for treatment advance. Here we developed an FGF4-driven human brainstem organoid model, which we used to genetically engineer H3.3K27M-altered DMG. We demonstrated that brainstem pontine glial specification is critical for DMG tumorigenesis, yielding infiltrative tumors that recapitulate patient-representative intratumoral heterogeneity. Prolonged GD2 chimeric antigen receptor (CAR) T cell treatment mirrored clinical outcomes and revealed extensive transcriptional heterogeneity, from which both potent effector and dysfunctional CAR T cell populations could be identified. Furthermore, incorporation of myeloid cells generated DMG-specific microglia that reduced treatment efficacy and revealed CAR T cell functional states most vulnerable to microglia-mediated immunosuppression. Thus, we present a representative DMG model offering a months-long experimental window in vitro, which we leveraged to delineate CAR T cell functionality and microglial impact, aiding therapy development for this devastating disease.
Authors
- Nils Bessler
- Celina Honhoff (ORCID: https://orcid.org/0000-0003-0167-3541)
- Annette Künkele (ORCID: https://orcid.org/0000-0002-8406-5412)
- Farid Keramati (ORCID: https://orcid.org/0000-0002-3034-4133)
- Sara Patrizi (ORCID: https://orcid.org/0000-0003-3133-8015)
- Anna Alemany (ORCID: https://orcid.org/0000-0002-0795-0290)
- Mariëtte E.G. Kranendonk (ORCID: https://orcid.org/0000-0001-9683-7494)
- Esmée J. van Vliet (ORCID: https://orcid.org/0000-0002-9301-1598)
- Ali Ertürk (ORCID: https://orcid.org/0000-0001-5163-5100)
- Sam de Blank (ORCID: https://orcid.org/0000-0003-2191-5675)
- Jurgen H. E. Kuball (ORCID: https://orcid.org/0000-0002-3914-7806)
- Annelisa M. Cornel (ORCID: https://orcid.org/0000-0001-8349-121X)
- Mario Barrera Román (ORCID: https://orcid.org/0000-0002-5919-2962)
- Cristian Ruiz Moreno
- Noëlle Dommann (ORCID: https://orcid.org/0000-0003-0475-0899)
- Christian Mayer (ORCID: https://orcid.org/0000-0003-3152-5574)
- María Alieva (ORCID: https://orcid.org/0000-0002-1425-9908)
- Raphaël Collot (ORCID: https://orcid.org/0000-0003-2080-1422)
- M. Kool
- Stefan Nierkens (ORCID: https://orcid.org/0000-0003-3406-817X)
- Mieke Roosen (ORCID: https://orcid.org/0000-0002-1880-5785)
- Anne C. Rios (ORCID: https://orcid.org/0000-0002-9082-8068)
- Daan J. Kloosterman (ORCID: https://orcid.org/0000-0003-1082-070X)
- Thijs J. M. van den Broek
- Ellen J Wehrens
- Evelina Miele
- Zsolt Sebestyén
- Hendrikus C. R. Ariese
- Hendrik G. Stunnenberg
- Amber K. L. Wezenaar
- Lucrezia C. D. E. Gatti
Institutions
- Radboud University Nijmegen (NL)
- German Cancer Research Center (DE)
- Utrecht University (NL)
- Heidelberg University (DE)
- Leiden University Medical Center (NL)
- Novo Nordisk Foundation (DK)
- Radboud University Medical Center (NL)
- Helmholtz Zentrum München (DE)
- University Medical Center Utrecht (NL)
- Humboldt-Universität zu Berlin (DE)
- Instituto de Investigaciones Biomédicas Sols-Morreale (ES)
- Radboud Institute for Molecular Life Sciences (NL)
- Oncode Institute (NL)
- Hopp Children's Cancer Center Heidelberg (DE)
- Bambino Gesù Children's Hospital (IT)
- Princess Máxima Center (NL)
- Max Planck Institute of Neurobiology (DE)
- Universidad Autónoma de Madrid (ES)
- Freie Universität Berlin (DE)
- Charité - Universitätsmedizin Berlin (DE)
Publication Details
- Journal
- Nature Cancer
- Published
- 2026-01-05
- DOI
- https://doi.org/10.1038/s43018-025-01084-0
- Citations
- 1
- Primary Topic
- CAR-T cell therapy research
- Type
- article
- Field-Weighted Citation Impact
- 24.20