Evaluating cytokine-based therapeutics in human 3D in vitro immune-containing assembloids
5 affiliations
- 1Roche Innovation Center Zurich, Roche Pharma Research and Early Development, F. Hoffmann-La Roche , ,
- 2Roche Innovation Center Basel, Roche Pharma Research and Early Development, F. Hoffmann-La Roche , ,
- 3Computational Sciences Center of Excellence, F. Hoffmann-La Roche , ,
- 4Institute of Human Biology (IHB), Roche Pharma Research and Early Development, F. Hoffmann-La Roche , ,
- 5Roche Innovation Center Munich, Roche Pharma Research and Early Development, F. Hoffmann-La Roche , ,
Abstract
Introduction
Cancer immunotherapies require accurate preclinical models that recapitulate the complex cellular interactions within the human tumor microenvironment. While patient-derived organoids have emerged as powerful tools for cancer research, standard cultures lack the stromal and immune compartments necessary to evaluate immunotherapeutic mechanisms. This limitation hampers the development of next-generation targeted cytokine therapies.Methods
We applied a 3D immune-containing colorectal cancer assembloid platform integrating patient-derived organoids, cancer-associated fibroblasts and allogeneic peripheral blood mononuclear cells within a collagen matrix. We characterized cellular responses to interferon-gamma (IFNg) and engineered IFNg receptor agonists targeted to fibroblast activation protein (FAP) through single-cell transcriptomics, immunohistochemistry, cytokine profiling, and functional T-cell migration assays.Results
The assembloid model successfully recapitulated key features of IFNg signaling, including tissue growth suppression, enhanced chemokine secretion (CXCL10), and CD8⁺ T-cell recruitment. Single-cell transcriptomics revealed that fibroblasts were the most responsive cell population, undergoing a phenotypic transition from myofibroblasts toward antigen-presenting CAFs (apCAFs), characterized by robust upregulation of MHC class II genes. FAP-targeted IFNg molecules exhibited attenuated activity compared to recombinant IFNg, successfully avoiding upregulation of the severe immunosuppressive checkpoint molecule PD-L1.Discussion
This 3D assembloid model provides a physiologically relevant platform for dissecting complex tumor-immune interactions and optimizing targeted immunotherapies. This approach enables the study of epithelial-stromal-immune crosstalk in cancer and offers a robust preclinical framework for screening and mechanistically validating next-generation targeted cytokine therapies.
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BibTeX
@article{maenhoudt2026evaluating,
title = {{Evaluating cytokine-based therapeutics in human 3D in vitro immune-containing assembloids}},
author = {Nina Maenhoudt and Mechthild Lütge and Melanie Obenloch and Aljona Caironi and Maria Kiratzis and Pol Jean-Mairet and Samuele Graziano and Philipp Fröbel and Tamara Hüsser and Emilio Yángüez and Maria Antsiferova and Adrian M. Filip and Christian Müller and Stephane Leclair and Stephan Gasser and Pablo Umaña and Sara Colombetti and Floriana Cremasco and Samuele Calabro},
journal = {Frontiers in cell and developmental biology},
year = {2026},
month = oct,
doi = {10.3389/fcell.2026.1930651},
url = {https://europepmc.org/article/PMC/PMC13717148},
note = {CC BY},
}
RIS
TY - JOUR
TI - Evaluating cytokine-based therapeutics in human 3D in vitro immune-containing assembloids
AU - Nina Maenhoudt
AU - Mechthild Lütge
AU - Melanie Obenloch
AU - Aljona Caironi
AU - Maria Kiratzis
AU - Pol Jean-Mairet
AU - Samuele Graziano
AU - Philipp Fröbel
AU - Tamara Hüsser
AU - Emilio Yángüez
AU - Maria Antsiferova
AU - Adrian M. Filip
AU - Christian Müller
AU - Stephane Leclair
AU - Stephan Gasser
AU - Pablo Umaña
AU - Sara Colombetti
AU - Floriana Cremasco
AU - Samuele Calabro
PY - 2026
DA - 2026/10/15
JO - Frontiers in cell and developmental biology
DO - 10.3389/fcell.2026.1930651
UR - https://europepmc.org/article/PMC/PMC13717148
AB - Introduction Cancer immunotherapies require accurate preclinical models that recapitulate the complex cellular interactions within the human tumor microenvironment. While patient-derived organoids have emerged as powerful tools for cancer research, standard cultures lack the stromal and immune compartments necessary to evaluate immunotherapeutic mechanisms. This limitation hampers the development of next-generation targeted cytokine therapies.Methods We applied a 3D immune-containing colorectal cancer assembloid platform integrating patient-derived organoids, cancer-associated fibroblasts and allogeneic peripheral blood mononuclear cells within a collagen matrix. We characterized cellular responses to interferon-gamma (IFNg) and engineered IFNg receptor agonists targeted to fibroblast activation protein (FAP) through single-cell transcriptomics, immunohistochemistry, cytokine profiling, and functional T-cell migration assays.Results The assembloid model successfully recapitulated key features of IFNg signaling, including tissue growth suppression, enhanced chemokine secretion (CXCL10), and CD8⁺ T-cell recruitment. Single-cell transcriptomics revealed that fibroblasts were the most responsive cell population, undergoing a phenotypic transition from myofibroblasts toward antigen-presenting CAFs (apCAFs), characterized by robust upregulation of MHC class II genes. FAP-targeted IFNg molecules exhibited attenuated activity compared to recombinant IFNg, successfully avoiding upregulation of the severe immunosuppressive checkpoint molecule PD-L1.Discussion This 3D assembloid model provides a physiologically relevant platform for dissecting complex tumor-immune interactions and optimizing targeted immunotherapies. This approach enables the study of epithelial-stromal-immune crosstalk in cancer and offers a robust preclinical framework for screening and mechanistically validating next-generation targeted cytokine therapies.
SN - 2296-634X
ER -