Cancer-Associated Fibroblasts Provide a Stromal Niche for Liver Cancer Organoids That Confers Trophic Effects and Therapy Resistance.

Animals Antineoplastic Agents / pharmacology Cancer-Associated Fibroblasts / metabolism Carcinoma, Hepatocellular / drug therapy Coculture Techniques Culture Media, Conditioned / metabolism Diethylnitrosamine / administration & dosage Drug Resistance, Neoplasm Humans Liver Neoplasms / drug therapy Liver Neoplasms, Experimental / chemically induced Mice Organoids / drug effects Paracrine Communication Primary Cell Culture Tumor Cells, Cultured Tumor Microenvironment Xenograft Model Antitumor Assays

Cell–Cell Contact Co-Culture Liver Tumor Organoids Paracrine Effect Stromal Cells

Journal

Cellular and molecular gastroenterology and hepatology

ISSN: 2352-345X

Titre abrégé: Cell Mol Gastroenterol Hepatol

Pays: United States

ID NLM: 101648302

Informations de publication

Date de publication:
2021

Historique:

received: 15 03 2020

revised: 05 09 2020

accepted: 08 09 2020

pubmed: 16 9 2020

medline: 21 12 2021

entrez: 15 9 2020

Statut: ppublish

Résumé

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. 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. 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. We successfully established murine and human 3D co-culture models and have shown robust effects of CAFs in liver cancer nurturing and treatment resistance.

Sections du résumé

BACKGROUND & AIMS

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.

Identifiants

DOI: 10.1016/j.jcmgh.2020.09.003 PMID: 32932015 PMC: PMC7788239

pubmed: 32932015

pii: S2352-345X(20)30140-5

doi: 10.1016/j.jcmgh.2020.09.003

pmc: PMC7788239

pii:

doi:

Substances chimiques

Antineoplastic Agents 0

Culture Media, Conditioned 0

Diethylnitrosamine 3IQ78TTX1A

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

Pagination

407-431