TY - JOUR
T1 - Dendritic cell phenotype and function in a 3D co-culture model of patient-derived metastatic colorectal cancer organoids
AU - Subtil, Beatriz
AU - Iyer, Kirti K.
AU - Poel, Dennis
AU - Bakkerus, Lotte
AU - Gorris, Mark A. J.
AU - Escalona, Jorge Cuenca
AU - Dries, Koen van den
AU - Cambi, Alessandra
AU - Verheul, Henk M. W.
AU - de Vries, I. Jolanda M.
AU - Tauriello, Daniele V. F.
N1 - Funding Information: DT is funded by a Hypatia Tenure Track Fellowship grant from the Radboudumc and by the Dutch Research Council(NWO/ZonMW VIDI grant number 91719371). IV is funded by EU grant Oncobiome (825410) and Health Holland/SGF grant DC4Balance (LSHM18056-SGF). Publisher Copyright: Copyright © 2023 Subtil, Iyer, Poel, Bakkerus, Gorris, Escalona, Dries, Cambi, Verheul, de Vries and Tauriello.
PY - 2023/1/25
Y1 - 2023/1/25
N2 - Colorectal cancer (CRC) remains one of the most aggressive and lethal cancers, with metastasis accounting for most deaths. As such, there is an unmet need for improved therapies for metastatic CRC (mCRC). Currently, the research focus is shifting towards the reciprocal interactions within the tumor microenvironment (TME), which prevent tumor clearance by the immune system. Dendritic cells (DCs) play a key role in the initiation and amplification of anti-tumor immune responses and in driving the clinical success of immunotherapies. Dissecting the interactions between DCs and CRC cells may open doors to identifying key mediators in tumor progression, and possible therapeutic targets. This requires representative, robust and versatile models and tools. Currently, there is a shortage of such in vitro systems to model the CRC TME and its tumor-immune cell interactions. Here we develop and establish a dynamic organotypic 3D co-culture system to recapitulate and untangle the interactions between DCs and patient-derived mCRC tumor organoids. To our knowledge, this is the first study investigating human DCs in co-culture with tumor organoids in a 3D, organotypic setting. This system reveals how mCRC organoids modulate and shape monocyte-derived DCs (MoDCs) behavior, phenotype, and function, within a collagen matrix, using techniques such as brightfield and fluorescence microscopy, flow cytometry, and fluorescence-activated cell sorting. Our 3D co-culture model shows high viability and extensive interaction between DCs and tumor organoids, and its structure resembles patient tissue sections. Furthermore, it is possible to retrieve DCs from the co-cultures and characterize their phenotypic and functional profile. In our study, the expression of activation markers in both mature and immature DCs and their ability to activate T cells were impacted by co-culture with tumor organoids. In the future, this direct co-culture platform can be adapted and exploited to study the CRC-DC interplay in more detail, enabling novel and broader insights into CRC-driven DC (dys)function.
AB - Colorectal cancer (CRC) remains one of the most aggressive and lethal cancers, with metastasis accounting for most deaths. As such, there is an unmet need for improved therapies for metastatic CRC (mCRC). Currently, the research focus is shifting towards the reciprocal interactions within the tumor microenvironment (TME), which prevent tumor clearance by the immune system. Dendritic cells (DCs) play a key role in the initiation and amplification of anti-tumor immune responses and in driving the clinical success of immunotherapies. Dissecting the interactions between DCs and CRC cells may open doors to identifying key mediators in tumor progression, and possible therapeutic targets. This requires representative, robust and versatile models and tools. Currently, there is a shortage of such in vitro systems to model the CRC TME and its tumor-immune cell interactions. Here we develop and establish a dynamic organotypic 3D co-culture system to recapitulate and untangle the interactions between DCs and patient-derived mCRC tumor organoids. To our knowledge, this is the first study investigating human DCs in co-culture with tumor organoids in a 3D, organotypic setting. This system reveals how mCRC organoids modulate and shape monocyte-derived DCs (MoDCs) behavior, phenotype, and function, within a collagen matrix, using techniques such as brightfield and fluorescence microscopy, flow cytometry, and fluorescence-activated cell sorting. Our 3D co-culture model shows high viability and extensive interaction between DCs and tumor organoids, and its structure resembles patient tissue sections. Furthermore, it is possible to retrieve DCs from the co-cultures and characterize their phenotypic and functional profile. In our study, the expression of activation markers in both mature and immature DCs and their ability to activate T cells were impacted by co-culture with tumor organoids. In the future, this direct co-culture platform can be adapted and exploited to study the CRC-DC interplay in more detail, enabling novel and broader insights into CRC-driven DC (dys)function.
KW - 3D co-culture
KW - dendritic cell dysfunction
KW - human dendritic cells
KW - immunosuppression
KW - metastatic colorectal cancer
KW - patient-derived tumor organoids
KW - tumor microenvironment
UR - http://www.scopus.com/inward/record.url?scp=85147565954&partnerID=8YFLogxK
U2 - https://doi.org/10.3389/fimmu.2023.1105244
DO - https://doi.org/10.3389/fimmu.2023.1105244
M3 - Article
C2 - 36761758
SN - 1664-3224
VL - 14
JO - Frontiers in immunology
JF - Frontiers in immunology
M1 - 1105244
ER -