A Novel Orthotopic Liver Cancer Model for Creating a Human-like Tumor Microenvironment

Abstract

Hepatocellular carcinoma (HCC) is the most common form of liver cancer. This study aims to develop a new method to generate an HCC mouse model with a human tumor, and imitates the tumor microenvironment (TME) of clinical patients. Here, we have generated functional, three-dimensional sheet-like human HCC organoids in vitro, using luciferase-expressing Huh7 cells, human iPSC-derived endothelial cells (iPSC-EC), and human iPSC-derived mesenchymal cells (iPSC-MC). The HCC organoid, capped by ultra-purified alginate gel, was implanted into the disrupted liver using an ultrasonic homogenizer in the immune-deficient mouse, which improved the survival and engraftment rate. We successfully introduced different types of controllable TME into the model and studied the roles of TME in HCC tumor growth. The results showed the role of the iPSC-EC and iPSC-MC combination, especially the iPSC-MC, in promoting HCC growth. We also demonstrated that liver fibrosis could promote HCC tumor growth. However, it is not affected by non-alcoholic fatty liver disease. Furthermore, the implantation of HCC organoids to humanized mice demonstrated that the immune response is important in slowing down tumor growth at an early stage. In conclusion, we have created an HCC model that is useful for studying HCC development and developing new treatment options in the future.

Combined Administration of Escitalopram Oxalate and Nivolumab Exhibits Synergistic Growth-Inhibitory Effects on Liver Cancer Cells through Inducing Apoptosis

“…Second, the current study used a subcutaneous xenograft tumor mouse model, which cannot accurately simulate the human tumor microenvironment. Therefore, better orthotopic liver cancer animal models, as described elsewhere [ 54 , 55 , 56 , 66 ], should be adopted in order to verify the effects of the combinational use of nivolumab and escitalopram oxalate.…”

Construction of tumor organoids and their application to cancer research and therapy

“…These characteristics made the co-culture models suitable for understanding and targeting the interactions between the immune niches and angiogenesis. In addition, some researchers have combined endothelial cells and other types of cells, such as mesenchymal cells and immune cells, for tumor organoid co-culture [38,46,47]. Various cell materials additionally supplemented for tumor organoid co-culture are summarized in Table 1.…”

Tumor organoids: applications in cancer modeling and potentials in precision medicine

“…Fibroblast growth factor receptor 2 (FGFR2) fusion proteins were reported to promote oncogenic transformation of mouse liver organoids to cholangiocarcinoma [93]. In another study, functional human HCC organoids were generated from Huh7 cells, human iPSC-derived mesenchymal cells (MCs), and human iPSC-derived endothelial cells (ECs) [94]. The combination of human iPSC-ECs and iPSC-MCs drove HCC growth, and the immune response was important for slowing tumor growth at an early stage [94].…”

“…In another study, functional human HCC organoids were generated from Huh7 cells, human iPSC-derived mesenchymal cells (MCs), and human iPSC-derived endothelial cells (ECs) [94]. The combination of human iPSC-ECs and iPSC-MCs drove HCC growth, and the immune response was important for slowing tumor growth at an early stage [94]. Sequential knockout and knock-in of driver mutations using CRISPR-Cas9 technology were conducted to generate genetically modified liver cancer organoids as a method to further investigate tumor pathogenesis [95].…”