1. Cancer-Associated Fibroblasts Provide a Stromal Niche for Liver Cancer Organoids That Confers Trophic Effects and Therapy Resistance
- Author
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Zhouhong Ge, Ron Smits, Lucia Campos Carrascosa, Dave Sprengers, Jaap Kwekkeboom, Jan N. M. IJzermans, Guoying Zhou, Maikel P. Peppelenbosch, Ruby Lieshout, Ling Wang, Wanlu Cao, Lisanne Noordam, Pengfei Li, Qin Yang, Junhong Su, Jiaye Liu, Luc J. W. van der Laan, Meng Li, Buyun Ma, Monique M A Verstegen, Qiuwei Pan, Ruyi Zhang, Gastroenterology & Hepatology, and Surgery
- Subjects
FAP, fibroblast-associated protein ,0301 basic medicine ,IGF, insulin-like growth factor ,Liver Tumor Organoids ,Wnt, wingless-related integration site ,Cell–Cell Contact ,Mice ,PCR, polymerase chain reaction ,Liver Neoplasms, Experimental ,0302 clinical medicine ,Cancer-Associated Fibroblasts ,Tumor Cells, Cultured ,Tumor Microenvironment ,Diethylnitrosamine ,Paracrine Effect ,OEM, organoids expansion medium ,Original Research ,AFP, α-fetoprotein ,Liver Neoplasms ,Gastroenterology ,ECM, extracellular matrix ,Organoids ,EpCAM, epithelial cell adhesion molecule ,NSG, NOD scid γ mouse ,FACS, fluorescence-activated cell sorter ,030211 gastroenterology & hepatology ,Liver cancer ,medicine.drug ,Sorafenib ,CCA, cholangiocarcinoma ,Carcinoma, Hepatocellular ,Stromal cell ,Liver tumor ,Primary Cell Culture ,PBS, phosphate-buffered saline ,Antineoplastic Agents ,Biology ,α-SMA, α-smooth muscle actin ,Co-Culture ,PDGFRA, platelet-derived growth factor receptor α ,03 medical and health sciences ,Paracrine signalling ,SDG 3 - Good Health and Well-being ,3D, 3-dimensional ,Paracrine Communication ,OBM, organoids basic medium ,medicine ,Animals ,Humans ,CAF, cancer-associated fibroblast ,EGF, epidermal growth factor ,Hepatology ,Cancer ,DEN, N-nitrosodiethylamine ,medicine.disease ,CSC, cancer stem cell ,Xenograft Model Antitumor Assays ,Coculture Techniques ,FGF, fibroblast growth factor ,IL, interleukin ,030104 developmental biology ,Drug Resistance, Neoplasm ,Culture Media, Conditioned ,Cancer cell ,Cancer research ,TCGA, The Cancer Genome Atlas ,DMEM, Dulbecco’s modified Eagle medium ,5-FU, 5-fluorouracil ,HGF, hepatocyte growth factor ,FCS, fetal calf serum ,Stromal Cells ,HCC, hepatocellular carcinoma - Abstract
Background & Aims Cancer-associated fibroblasts (CAFs) play a key role in the cancer process, but the research progress is hampered by the paucity of preclinical models that are essential for mechanistic dissection of cancer cell–CAF interactions. Here, we aimed to establish 3-dimensional (3D) organotypic co-cultures of primary liver tumor–derived organoids with CAFs, and to understand their interactions and the response to treatment. Methods Liver tumor organoids and CAFs were cultured from murine and human primary liver tumors. 3D co-culture models of tumor organoids with CAFs and Transwell culture systems were established in vitro. A xenograft model was used to investigate the cell–cell interactions in vivo. Gene expression analysis of CAF markers in our hepatocellular carcinoma cohort and an online liver cancer database indicated the clinical relevance of CAFs. Results To functionally investigate the interactions of liver cancer cells with CAFs, we successfully established murine and human 3D co-culture models of liver tumor organoids with CAFs. CAFs promoted tumor organoid growth in co-culture with direct cell–cell contact and in a Transwell system via paracrine signaling. Vice versa, cancer cells secrete paracrine factors regulating CAF physiology. Co-transplantation of CAFs with liver tumor organoids of mouse or human origin promoted tumor growth in xenograft models. Moreover, tumor organoids conferred resistance to clinically used anticancer drugs including sorafenib, regorafenib, and 5-fluorouracil in the presence of CAFs, or the conditioned medium of CAFs. Conclusions We successfully established murine and human 3D co-culture models and have shown robust effects of CAFs in liver cancer nurturing and treatment resistance., Graphical abstract
- Published
- 2021
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