Cancer‐associated fibroblasts nurture LGR5 marked liver tumor‐initiating cells and promote their tumor formation, growth, and metastasis
Abstract Background & Aims In liver cancer, leucine‐rich repeat‐containing G‐protein coupled receptor 5 (LGR5) compartment represents an important tumor‐initiating cell (TIC) population and served as a potential therapeutic target. Cancer‐associated fibroblasts (CAFs) is a critical part of the t...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2023-09-01
|
| Series: | Cancer Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/cam4.6408 |
| _version_ | 1797672357773443072 |
|---|---|
| author | Mingna Zhang Yiqiao Fang Xia Fu Jiaye Liu Yang Liu Zhounan Zhu Yinyun Ni Menglin Yao Qiuwei Pan Wanlu Cao Zhihui Li Chunyan Dong |
| author_facet | Mingna Zhang Yiqiao Fang Xia Fu Jiaye Liu Yang Liu Zhounan Zhu Yinyun Ni Menglin Yao Qiuwei Pan Wanlu Cao Zhihui Li Chunyan Dong |
| author_sort | Mingna Zhang |
| collection | DOAJ |
| description | Abstract Background & Aims In liver cancer, leucine‐rich repeat‐containing G‐protein coupled receptor 5 (LGR5) compartment represents an important tumor‐initiating cell (TIC) population and served as a potential therapeutic target. Cancer‐associated fibroblasts (CAFs) is a critical part of the tumor microenvironment, heavily influenced TIC function and fate. However, deeply investigations have been hindered by the lack of accurate preclinical models to investigate the interaction between CAFs and TIC. Organoids model have achieved major advancements as a precious research model for recapitulating the morphological aspects of organs, and thus also serving as a candidate model to investigate the mutual interaction between different cell types. Consequently, this study aimed to construct a three‐dimensional (3D) co‐culture organoid model of primary LGR5‐expressing tumor stem cells from primary murine liver tumors with CAFs to investigate the impact of CAFs on LGR5 marked TICs in liver cancer. Materials and Methods First, both of the transgenic LGR5‐diphtheria toxin receptor (DTR)‐GFP knock‐in mice and transgenic Rosa26‐mT mice developed primary liver tumors by diethylnitrosamine (DEN) administration. Tumor organoids and CAFs were generated from those primary liver cancer separately. Second, LGR5‐expressing TICs organoid with CAFs were established ex vivo based on cell–cell contact or trans‐well co‐culture system, and the mutual influence between those two types of cells was further investigated. Subsequently, immunodeficient mouse‐based xenograft model was further adopted to evaluate the influence of CAFs to LGR5 tumor stem cell, tumor formation, and metastasis. Results The co‐culture organoid model composed of murine liver tumor LGR5+ tumor‐initiating cells and CAFs in 3D co‐culture was successfully established, with the intention to investigate their mutual interaction. The existence of CAFs upon engrafting tumor organoids resulted in dramatic higher number of LGR5+ cells in the neoplasia when compared with engrafting tumor organoids alone. Furthermore, ex vivo culture of isolated LGR5+ cells from tumors of co‐engrafted mice formed significantly larger size of organoids than mono‐engrafted. Our results also indicated significantly larger size and number of formed organoids, when LGR5+ cells co‐cultured with CAF in both cell–cell contact and paracrine signaling in vitro, comparing to LGR5+ cells alone. Furthermore, we found that specific knockout of LGR5 expressing cells suppressed CAF‐mediated promotion of tumor formation, growth, and metastasis in the experimental mice model. Conclusions Altogether, in a 3D co‐culture type of murine liver LGR5+ cells and cancer‐associated fibroblasts, we have demonstrated robust effects of CAFs in the promotion of LGR5 marked liver TICs. We also further revealed the influence of tumor microenvironment on stem cell‐related therapy, suggesting the possibility of combing CAF‐targeted and tumor stem cell targeted therapy in treating liver cancer. |
| first_indexed | 2024-03-11T21:28:47Z |
| format | Article |
| id | doaj.art-6dca15fe43bb4954acd3cd23f077e098 |
| institution | Directory Open Access Journal |
| issn | 2045-7634 |
| language | English |
| last_indexed | 2024-03-11T21:28:47Z |
| publishDate | 2023-09-01 |
| publisher | Wiley |
| record_format | Article |
| series | Cancer Medicine |
| spelling | doaj.art-6dca15fe43bb4954acd3cd23f077e0982023-09-27T11:46:08ZengWileyCancer Medicine2045-76342023-09-011217180321804910.1002/cam4.6408Cancer‐associated fibroblasts nurture LGR5 marked liver tumor‐initiating cells and promote their tumor formation, growth, and metastasisMingna Zhang0Yiqiao Fang1Xia Fu2Jiaye Liu3Yang Liu4Zhounan Zhu5Yinyun Ni6Menglin Yao7Qiuwei Pan8Wanlu Cao9Zhihui Li10Chunyan Dong11Department of Oncology Postgraduate Training Base of Jinzhou Medical University, Shanghai East Hospital Shanghai ChinaDepartment of Thyroid and Parathyroid Surgery, West China Hospital Sichuan University Chengdu Sichuan ChinaDepartment of Outpatients, West China Hospital Sichuan University Chengdu Sichuan ChinaDepartment of Thyroid and Parathyroid Surgery, West China Hospital Sichuan University Chengdu Sichuan ChinaDepartment of Obsterics and Gynecology, Second Affiliated Hospital Chongqing Medical University Chongqing ChinaDepartment of Oncology Shanghai East Hospital, Tongji University School of Medicine, Tongji University Shanghai People's Republic of ChinaDepartment of Respiratory and Critical Care Medicine, National Clinic al Research Center for Geriatrics, Center of Precision Medicine, Precision Medicine Key Laboratory of Sichuan Province, Frontiers Science Center for Disease‐related Molecular Network, West China Hospital, West China School of Medicine Sichuan University Chengdu Sichuan ChinaDepartment of Respiratory and Critical Care Medicine, National Clinic al Research Center for Geriatrics, Center of Precision Medicine, Precision Medicine Key Laboratory of Sichuan Province, Frontiers Science Center for Disease‐related Molecular Network, West China Hospital, West China School of Medicine Sichuan University Chengdu Sichuan ChinaDepartment of Gastroenterology and Hepatology Erasmus Medical Center Rotterdam the NetherlandsDepartment of Oncology Shanghai East Hospital, Tongji University School of Medicine, Tongji University Shanghai People's Republic of ChinaDepartment of Thyroid and Parathyroid Surgery, West China Hospital Sichuan University Chengdu Sichuan ChinaDepartment of Oncology Postgraduate Training Base of Jinzhou Medical University, Shanghai East Hospital Shanghai ChinaAbstract Background & Aims In liver cancer, leucine‐rich repeat‐containing G‐protein coupled receptor 5 (LGR5) compartment represents an important tumor‐initiating cell (TIC) population and served as a potential therapeutic target. Cancer‐associated fibroblasts (CAFs) is a critical part of the tumor microenvironment, heavily influenced TIC function and fate. However, deeply investigations have been hindered by the lack of accurate preclinical models to investigate the interaction between CAFs and TIC. Organoids model have achieved major advancements as a precious research model for recapitulating the morphological aspects of organs, and thus also serving as a candidate model to investigate the mutual interaction between different cell types. Consequently, this study aimed to construct a three‐dimensional (3D) co‐culture organoid model of primary LGR5‐expressing tumor stem cells from primary murine liver tumors with CAFs to investigate the impact of CAFs on LGR5 marked TICs in liver cancer. Materials and Methods First, both of the transgenic LGR5‐diphtheria toxin receptor (DTR)‐GFP knock‐in mice and transgenic Rosa26‐mT mice developed primary liver tumors by diethylnitrosamine (DEN) administration. Tumor organoids and CAFs were generated from those primary liver cancer separately. Second, LGR5‐expressing TICs organoid with CAFs were established ex vivo based on cell–cell contact or trans‐well co‐culture system, and the mutual influence between those two types of cells was further investigated. Subsequently, immunodeficient mouse‐based xenograft model was further adopted to evaluate the influence of CAFs to LGR5 tumor stem cell, tumor formation, and metastasis. Results The co‐culture organoid model composed of murine liver tumor LGR5+ tumor‐initiating cells and CAFs in 3D co‐culture was successfully established, with the intention to investigate their mutual interaction. The existence of CAFs upon engrafting tumor organoids resulted in dramatic higher number of LGR5+ cells in the neoplasia when compared with engrafting tumor organoids alone. Furthermore, ex vivo culture of isolated LGR5+ cells from tumors of co‐engrafted mice formed significantly larger size of organoids than mono‐engrafted. Our results also indicated significantly larger size and number of formed organoids, when LGR5+ cells co‐cultured with CAF in both cell–cell contact and paracrine signaling in vitro, comparing to LGR5+ cells alone. Furthermore, we found that specific knockout of LGR5 expressing cells suppressed CAF‐mediated promotion of tumor formation, growth, and metastasis in the experimental mice model. Conclusions Altogether, in a 3D co‐culture type of murine liver LGR5+ cells and cancer‐associated fibroblasts, we have demonstrated robust effects of CAFs in the promotion of LGR5 marked liver TICs. We also further revealed the influence of tumor microenvironment on stem cell‐related therapy, suggesting the possibility of combing CAF‐targeted and tumor stem cell targeted therapy in treating liver cancer.https://doi.org/10.1002/cam4.6408cancer‐associated fibroblastsdiphtheria toxin receptorleucine‐rich repeat‐containing G‐protein coupled receptor 5liver cancertumor‐initiating cells |
| spellingShingle | Mingna Zhang Yiqiao Fang Xia Fu Jiaye Liu Yang Liu Zhounan Zhu Yinyun Ni Menglin Yao Qiuwei Pan Wanlu Cao Zhihui Li Chunyan Dong Cancer‐associated fibroblasts nurture LGR5 marked liver tumor‐initiating cells and promote their tumor formation, growth, and metastasis Cancer Medicine cancer‐associated fibroblasts diphtheria toxin receptor leucine‐rich repeat‐containing G‐protein coupled receptor 5 liver cancer tumor‐initiating cells |
| title | Cancer‐associated fibroblasts nurture LGR5 marked liver tumor‐initiating cells and promote their tumor formation, growth, and metastasis |
| title_full | Cancer‐associated fibroblasts nurture LGR5 marked liver tumor‐initiating cells and promote their tumor formation, growth, and metastasis |
| title_fullStr | Cancer‐associated fibroblasts nurture LGR5 marked liver tumor‐initiating cells and promote their tumor formation, growth, and metastasis |
| title_full_unstemmed | Cancer‐associated fibroblasts nurture LGR5 marked liver tumor‐initiating cells and promote their tumor formation, growth, and metastasis |
| title_short | Cancer‐associated fibroblasts nurture LGR5 marked liver tumor‐initiating cells and promote their tumor formation, growth, and metastasis |
| title_sort | cancer associated fibroblasts nurture lgr5 marked liver tumor initiating cells and promote their tumor formation growth and metastasis |
| topic | cancer‐associated fibroblasts diphtheria toxin receptor leucine‐rich repeat‐containing G‐protein coupled receptor 5 liver cancer tumor‐initiating cells |
| url | https://doi.org/10.1002/cam4.6408 |
| work_keys_str_mv | AT mingnazhang cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis AT yiqiaofang cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis AT xiafu cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis AT jiayeliu cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis AT yangliu cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis AT zhounanzhu cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis AT yinyunni cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis AT menglinyao cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis AT qiuweipan cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis AT wanlucao cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis AT zhihuili cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis AT chunyandong cancerassociatedfibroblastsnurturelgr5markedlivertumorinitiatingcellsandpromotetheirtumorformationgrowthandmetastasis |