Co-cultures of colon cancer cells and cancer-associated fibroblasts recapitulate the aggressive features of mesenchymal-like colon cancer
BackgroundPoor prognosis in colon cancer is associated with a high content of cancer-associated fibroblasts (CAFs) and an immunosuppressive tumor microenvironment. The relationship between these two features is incompletely understood. Here, we aimed to generate a model system for studying the inter...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-05-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1053920/full |
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| author | Esther Strating Mathijs P. Verhagen Emerens Wensink Ester Dünnebach Ester Dünnebach Liza Wijler Itziar Aranguren Alberto Sanchez De la Cruz Niek A. Peters Joris H. Hageman Mirjam M. C. van der Net Susanne van Schelven Jamila Laoukili Riccardo Fodde Jeanine Roodhart Stefan Nierkens Stefan Nierkens Hugo Snippert Martijn Gloerich Inne Borel Rinkes Sjoerd G. Elias Onno Kranenburg Onno Kranenburg |
| author_facet | Esther Strating Mathijs P. Verhagen Emerens Wensink Ester Dünnebach Ester Dünnebach Liza Wijler Itziar Aranguren Alberto Sanchez De la Cruz Niek A. Peters Joris H. Hageman Mirjam M. C. van der Net Susanne van Schelven Jamila Laoukili Riccardo Fodde Jeanine Roodhart Stefan Nierkens Stefan Nierkens Hugo Snippert Martijn Gloerich Inne Borel Rinkes Sjoerd G. Elias Onno Kranenburg Onno Kranenburg |
| author_sort | Esther Strating |
| collection | DOAJ |
| description | BackgroundPoor prognosis in colon cancer is associated with a high content of cancer-associated fibroblasts (CAFs) and an immunosuppressive tumor microenvironment. The relationship between these two features is incompletely understood. Here, we aimed to generate a model system for studying the interaction between cancer cells and CAFs and their effect on immune-related cytokines and T cell proliferation.MethodsCAFs were isolated from colon cancer liver metastases and were immortalized to prolong lifespan and improve robustness and reproducibility. Established medium and matrix compositions that support the growth of patient-derived organoids were adapted to also support CAF growth. Changes in growth pattern and cellular re-organization were assessed by confocal microscopy, live cell imaging, and immunofluorescence. Single cell RNA sequencing was used to study CAF/organoid co-culture-induced phenotypic changes in both cell types. Conditioned media were used to quantify the production of immunosuppressive factors and to assess their effect on T cell proliferation.ResultsWe developed a co-culture system in which colon cancer organoids and CAFs spontaneously organize into superstructures with a high capacity to contract and stiffen the extracellular matrix (ECM). CAF-produced collagen IV provided a basement membrane supporting cancer cell organization into glandular structures, reminiscent of human cancer histology. Single cell RNA sequencing analysis showed that CAFs induced a partial epithelial-to-mesenchymal-transition in a subpopulation of cancer cells, similar to what is observed in the mesenchymal-like consensus molecular subtype 4 (CMS4) colon cancer. CAFs in co-culture were characterized by high expression of ECM components, ECM-remodeling enzymes, glycolysis, hypoxia, and genes involved in immunosuppression. An expression signature derived from CAFs in co-culture identified a subpopulation of glycolytic myofibroblasts specifically residing in CMS1 and CMS4 colon cancer. Medium conditioned by co-cultures contained high levels of the immunosuppressive factors TGFβ1, VEGFA and lactate, and potently inhibited T cell proliferation.ConclusionCo-cultures of organoids and immortalized CAFs recapitulate the histological, biophysical, and immunosuppressive features of aggressive mesenchymal-like human CRC. The model can be used to study the mechanisms of immunosuppression and to test therapeutic strategies targeting the cross-talk between CAFs and cancer cells. It can be further modified to represent distinct colon cancer subtypes and (organ-specific) microenvironments. |
| first_indexed | 2024-04-09T12:30:47Z |
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| id | doaj.art-90ab2b9cb2c4478f87f361dda37eab04 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-09T12:30:47Z |
| publishDate | 2023-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-90ab2b9cb2c4478f87f361dda37eab042023-05-16T05:02:03ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-05-011410.3389/fimmu.2023.10539201053920Co-cultures of colon cancer cells and cancer-associated fibroblasts recapitulate the aggressive features of mesenchymal-like colon cancerEsther Strating0Mathijs P. Verhagen1Emerens Wensink2Ester Dünnebach3Ester Dünnebach4Liza Wijler5Itziar Aranguren6Alberto Sanchez De la Cruz7Niek A. Peters8Joris H. Hageman9Mirjam M. C. van der Net10Susanne van Schelven11Jamila Laoukili12Riccardo Fodde13Jeanine Roodhart14Stefan Nierkens15Stefan Nierkens16Hugo Snippert17Martijn Gloerich18Inne Borel Rinkes19Sjoerd G. Elias20Onno Kranenburg21Onno Kranenburg22Laboratory Translational Oncology, Division of Imaging and Cancer, University Medical Center Utrecht, Utrecht, NetherlandsDepartment of Pathology, Erasmus Medical Center, Rotterdam, NetherlandsDepartment of Medical Oncology, Division of Imaging and Cancer, University Medical Center Utrecht, Utrecht, NetherlandsCenter for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, NetherlandsPrincess Máxima Center for Pediatric Oncology, Utrecht, NetherlandsLaboratory Translational Oncology, Division of Imaging and Cancer, University Medical Center Utrecht, Utrecht, NetherlandsLaboratory Translational Oncology, Division of Imaging and Cancer, University Medical Center Utrecht, Utrecht, NetherlandsLaboratory Translational Oncology, Division of Imaging and Cancer, University Medical Center Utrecht, Utrecht, NetherlandsLaboratory Translational Oncology, Division of Imaging and Cancer, University Medical Center Utrecht, Utrecht, NetherlandsCenter for Molecular Medicine, Division LAB, University Medical Center Utrecht, Utrecht, NetherlandsCenter for Molecular Medicine, Division LAB, University Medical Center Utrecht, Utrecht, NetherlandsLaboratory Translational Oncology, Division of Imaging and Cancer, University Medical Center Utrecht, Utrecht, NetherlandsLaboratory Translational Oncology, Division of Imaging and Cancer, University Medical Center Utrecht, Utrecht, NetherlandsDepartment of Pathology, Erasmus Medical Center, Rotterdam, NetherlandsDepartment of Medical Oncology, Division of Imaging and Cancer, University Medical Center Utrecht, Utrecht, NetherlandsCenter for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, NetherlandsPrincess Máxima Center for Pediatric Oncology, Utrecht, NetherlandsCenter for Molecular Medicine, Division LAB, University Medical Center Utrecht, Utrecht, NetherlandsCenter for Molecular Medicine, Division LAB, University Medical Center Utrecht, Utrecht, NetherlandsLaboratory Translational Oncology, Division of Imaging and Cancer, University Medical Center Utrecht, Utrecht, NetherlandsDepartment of Epidemiology, Julius Center for Health Sciences and Primary Care, University Medical Center Utrecht, Utrecht, NetherlandsLaboratory Translational Oncology, Division of Imaging and Cancer, University Medical Center Utrecht, Utrecht, NetherlandsUtrecht Platform for Organoid Technology, Utrecht University, Utrecht, NetherlandsBackgroundPoor prognosis in colon cancer is associated with a high content of cancer-associated fibroblasts (CAFs) and an immunosuppressive tumor microenvironment. The relationship between these two features is incompletely understood. Here, we aimed to generate a model system for studying the interaction between cancer cells and CAFs and their effect on immune-related cytokines and T cell proliferation.MethodsCAFs were isolated from colon cancer liver metastases and were immortalized to prolong lifespan and improve robustness and reproducibility. Established medium and matrix compositions that support the growth of patient-derived organoids were adapted to also support CAF growth. Changes in growth pattern and cellular re-organization were assessed by confocal microscopy, live cell imaging, and immunofluorescence. Single cell RNA sequencing was used to study CAF/organoid co-culture-induced phenotypic changes in both cell types. Conditioned media were used to quantify the production of immunosuppressive factors and to assess their effect on T cell proliferation.ResultsWe developed a co-culture system in which colon cancer organoids and CAFs spontaneously organize into superstructures with a high capacity to contract and stiffen the extracellular matrix (ECM). CAF-produced collagen IV provided a basement membrane supporting cancer cell organization into glandular structures, reminiscent of human cancer histology. Single cell RNA sequencing analysis showed that CAFs induced a partial epithelial-to-mesenchymal-transition in a subpopulation of cancer cells, similar to what is observed in the mesenchymal-like consensus molecular subtype 4 (CMS4) colon cancer. CAFs in co-culture were characterized by high expression of ECM components, ECM-remodeling enzymes, glycolysis, hypoxia, and genes involved in immunosuppression. An expression signature derived from CAFs in co-culture identified a subpopulation of glycolytic myofibroblasts specifically residing in CMS1 and CMS4 colon cancer. Medium conditioned by co-cultures contained high levels of the immunosuppressive factors TGFβ1, VEGFA and lactate, and potently inhibited T cell proliferation.ConclusionCo-cultures of organoids and immortalized CAFs recapitulate the histological, biophysical, and immunosuppressive features of aggressive mesenchymal-like human CRC. The model can be used to study the mechanisms of immunosuppression and to test therapeutic strategies targeting the cross-talk between CAFs and cancer cells. It can be further modified to represent distinct colon cancer subtypes and (organ-specific) microenvironments.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1053920/fullcolorectal cancerCMS4immunosuppressivemicroenvironmentcancer-associated fibroblast (CAF) |
| spellingShingle | Esther Strating Mathijs P. Verhagen Emerens Wensink Ester Dünnebach Ester Dünnebach Liza Wijler Itziar Aranguren Alberto Sanchez De la Cruz Niek A. Peters Joris H. Hageman Mirjam M. C. van der Net Susanne van Schelven Jamila Laoukili Riccardo Fodde Jeanine Roodhart Stefan Nierkens Stefan Nierkens Hugo Snippert Martijn Gloerich Inne Borel Rinkes Sjoerd G. Elias Onno Kranenburg Onno Kranenburg Co-cultures of colon cancer cells and cancer-associated fibroblasts recapitulate the aggressive features of mesenchymal-like colon cancer Frontiers in Immunology colorectal cancer CMS4 immunosuppressive microenvironment cancer-associated fibroblast (CAF) |
| title | Co-cultures of colon cancer cells and cancer-associated fibroblasts recapitulate the aggressive features of mesenchymal-like colon cancer |
| title_full | Co-cultures of colon cancer cells and cancer-associated fibroblasts recapitulate the aggressive features of mesenchymal-like colon cancer |
| title_fullStr | Co-cultures of colon cancer cells and cancer-associated fibroblasts recapitulate the aggressive features of mesenchymal-like colon cancer |
| title_full_unstemmed | Co-cultures of colon cancer cells and cancer-associated fibroblasts recapitulate the aggressive features of mesenchymal-like colon cancer |
| title_short | Co-cultures of colon cancer cells and cancer-associated fibroblasts recapitulate the aggressive features of mesenchymal-like colon cancer |
| title_sort | co cultures of colon cancer cells and cancer associated fibroblasts recapitulate the aggressive features of mesenchymal like colon cancer |
| topic | colorectal cancer CMS4 immunosuppressive microenvironment cancer-associated fibroblast (CAF) |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1053920/full |
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