Evidence of steady-state fibroblast subtypes in the normal human breast as cells-of-origin for perturbed-state fibroblasts in breast cancer
Abstract Background Human breast cancer most frequently originates within a well-defined anatomical structure referred to as the terminal duct lobular unit (TDLU). This structure is endowed with its very own lobular fibroblasts representing one out of two steady-state fibroblast subtypes—the other b...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-01-01
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| Series: | Breast Cancer Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13058-024-01763-3 |
| _version_ | 1797349664656195584 |
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| author | Mikkel Morsing Bagger Jonas Sjölund Jiyoung Kim Katharina Theresa Kohler René Villadsen Abbas Jafari Moustapha Kassem Kristian Pietras Lone Rønnov-Jessen Ole William Petersen |
| author_facet | Mikkel Morsing Bagger Jonas Sjölund Jiyoung Kim Katharina Theresa Kohler René Villadsen Abbas Jafari Moustapha Kassem Kristian Pietras Lone Rønnov-Jessen Ole William Petersen |
| author_sort | Mikkel Morsing Bagger |
| collection | DOAJ |
| description | Abstract Background Human breast cancer most frequently originates within a well-defined anatomical structure referred to as the terminal duct lobular unit (TDLU). This structure is endowed with its very own lobular fibroblasts representing one out of two steady-state fibroblast subtypes—the other being interlobular fibroblasts. While cancer-associated fibroblasts (CAFs) are increasingly appreciated as covering a spectrum of perturbed states, we lack a coherent understanding of their relationship—if any—with the steady-state fibroblast subtypes. To address this, we here established two autologous CAF lines representing inflammatory CAFs (iCAFs) and myofibroblast CAFs (myCAFs) and compared them with already established interlobular- and lobular fibroblasts with respect to their origin and impact on tumor formation. Methods Primary breast tumor-derived CAFs were transduced to express human telomerase reverse transcriptase (hTERT) and sorted into CD105low and CD105high populations using fluorescence-activated cell sorting (FACS). The two populations were tested for differentiation similarities to iCAF and myCAF states through transcriptome-wide RNA-Sequencing (RNA-Seq) including comparison to an available iCAF-myCAF cell state atlas. Inference of origin in interlobular and lobular fibroblasts relied on RNA-Seq profiles, immunocytochemistry and growth characteristics. Osteogenic differentiation and bone formation assays in culture and in vivo were employed to gauge for origin in bone marrow-derived mesenchymal stem cells (bMSCs). Functional characteristics were assessed with respect to contractility in culture and interaction with tumor cells in mouse xenografts. The cells’ gene expression signatures were tested for association with clinical outcome of breast cancer patients using survival data from The Cancer Genome Atlas database. Results We demonstrate that iCAFs have properties in common with interlobular fibroblasts while myCAFs and lobular fibroblasts are related. None of the CAFs qualify as bMSCs as revealed by lack of critical performance in bone formation assays. Functionally, myCAFs and lobular fibroblasts are almost equally tumor promoting as opposed to iCAFs and interlobular fibroblasts. A myCAF gene signature is found to associate with poor breast cancer-specific survival. Conclusions We propose that iCAFs and myCAFs originate in interlobular and lobular fibroblasts, respectively, and more importantly, that the tumor-promoting properties of lobular fibroblasts render the TDLU an epicenter for breast cancer evolution. |
| first_indexed | 2024-03-08T12:33:40Z |
| format | Article |
| id | doaj.art-a46f3b614fcd4ea7b71577a94fffc26e |
| institution | Directory Open Access Journal |
| issn | 1465-542X |
| language | English |
| last_indexed | 2024-03-08T12:33:40Z |
| publishDate | 2024-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Breast Cancer Research |
| spelling | doaj.art-a46f3b614fcd4ea7b71577a94fffc26e2024-01-21T12:40:22ZengBMCBreast Cancer Research1465-542X2024-01-0126111910.1186/s13058-024-01763-3Evidence of steady-state fibroblast subtypes in the normal human breast as cells-of-origin for perturbed-state fibroblasts in breast cancerMikkel Morsing Bagger0Jonas Sjölund1Jiyoung Kim2Katharina Theresa Kohler3René Villadsen4Abbas Jafari5Moustapha Kassem6Kristian Pietras7Lone Rønnov-Jessen8Ole William Petersen9Division of Translational Cancer Research, Department of Laboratory Medicine, Lund University Cancer Centre, Lund UniversityDivision of Translational Cancer Research, Department of Laboratory Medicine, Lund University Cancer Centre, Lund UniversityDepartment of Cellular and Molecular Medicine, University of CopenhagenDepartment of Cellular and Molecular Medicine, University of CopenhagenDepartment of Cellular and Molecular Medicine, University of CopenhagenDepartment of Cellular and Molecular Medicine, University of CopenhagenDepartment of Cellular and Molecular Medicine, University of CopenhagenDivision of Translational Cancer Research, Department of Laboratory Medicine, Lund University Cancer Centre, Lund UniversitySection for Cell Biology and Physiology, Department of Biology, University of CopenhagenDepartment of Cellular and Molecular Medicine, University of CopenhagenAbstract Background Human breast cancer most frequently originates within a well-defined anatomical structure referred to as the terminal duct lobular unit (TDLU). This structure is endowed with its very own lobular fibroblasts representing one out of two steady-state fibroblast subtypes—the other being interlobular fibroblasts. While cancer-associated fibroblasts (CAFs) are increasingly appreciated as covering a spectrum of perturbed states, we lack a coherent understanding of their relationship—if any—with the steady-state fibroblast subtypes. To address this, we here established two autologous CAF lines representing inflammatory CAFs (iCAFs) and myofibroblast CAFs (myCAFs) and compared them with already established interlobular- and lobular fibroblasts with respect to their origin and impact on tumor formation. Methods Primary breast tumor-derived CAFs were transduced to express human telomerase reverse transcriptase (hTERT) and sorted into CD105low and CD105high populations using fluorescence-activated cell sorting (FACS). The two populations were tested for differentiation similarities to iCAF and myCAF states through transcriptome-wide RNA-Sequencing (RNA-Seq) including comparison to an available iCAF-myCAF cell state atlas. Inference of origin in interlobular and lobular fibroblasts relied on RNA-Seq profiles, immunocytochemistry and growth characteristics. Osteogenic differentiation and bone formation assays in culture and in vivo were employed to gauge for origin in bone marrow-derived mesenchymal stem cells (bMSCs). Functional characteristics were assessed with respect to contractility in culture and interaction with tumor cells in mouse xenografts. The cells’ gene expression signatures were tested for association with clinical outcome of breast cancer patients using survival data from The Cancer Genome Atlas database. Results We demonstrate that iCAFs have properties in common with interlobular fibroblasts while myCAFs and lobular fibroblasts are related. None of the CAFs qualify as bMSCs as revealed by lack of critical performance in bone formation assays. Functionally, myCAFs and lobular fibroblasts are almost equally tumor promoting as opposed to iCAFs and interlobular fibroblasts. A myCAF gene signature is found to associate with poor breast cancer-specific survival. Conclusions We propose that iCAFs and myCAFs originate in interlobular and lobular fibroblasts, respectively, and more importantly, that the tumor-promoting properties of lobular fibroblasts render the TDLU an epicenter for breast cancer evolution.https://doi.org/10.1186/s13058-024-01763-3Breast cancerCancer-associated fibroblastFibroblastiCAFmyCAFCell line |
| spellingShingle | Mikkel Morsing Bagger Jonas Sjölund Jiyoung Kim Katharina Theresa Kohler René Villadsen Abbas Jafari Moustapha Kassem Kristian Pietras Lone Rønnov-Jessen Ole William Petersen Evidence of steady-state fibroblast subtypes in the normal human breast as cells-of-origin for perturbed-state fibroblasts in breast cancer Breast Cancer Research Breast cancer Cancer-associated fibroblast Fibroblast iCAF myCAF Cell line |
| title | Evidence of steady-state fibroblast subtypes in the normal human breast as cells-of-origin for perturbed-state fibroblasts in breast cancer |
| title_full | Evidence of steady-state fibroblast subtypes in the normal human breast as cells-of-origin for perturbed-state fibroblasts in breast cancer |
| title_fullStr | Evidence of steady-state fibroblast subtypes in the normal human breast as cells-of-origin for perturbed-state fibroblasts in breast cancer |
| title_full_unstemmed | Evidence of steady-state fibroblast subtypes in the normal human breast as cells-of-origin for perturbed-state fibroblasts in breast cancer |
| title_short | Evidence of steady-state fibroblast subtypes in the normal human breast as cells-of-origin for perturbed-state fibroblasts in breast cancer |
| title_sort | evidence of steady state fibroblast subtypes in the normal human breast as cells of origin for perturbed state fibroblasts in breast cancer |
| topic | Breast cancer Cancer-associated fibroblast Fibroblast iCAF myCAF Cell line |
| url | https://doi.org/10.1186/s13058-024-01763-3 |
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