A Novel Cell Line Based Orthotopic Xenograft Mouse Model That Recapitulates Human Hepatoblastoma

Abstract

Currently, preclinical testing of therapies for hepatoblastoma (HB) is limited to subcutaneous and intrasplenic xenograft models that do not recapitulate the hepatic tumors seen in patients. We hypothesized that injection of HB cell lines into the livers of mice would result in liver tumors that resemble their clinical counterparts. HepG2 and Huh-6 HB cell lines were injected, and tumor growth was monitored with bioluminescence imaging (BLI) and magnetic resonance imaging (MRI). Levels of human α-fetoprotein (AFP) were monitored in the serum of animals. Immunohistochemical and gene expression analyses were also completed on xenograft tumor samples. BLI signal indicative of tumor growth was seen in 55% of HepG2- and Huh-6-injected animals after a period of four to seven weeks. Increased AFP levels correlated with tumor growth. MRI showed large intrahepatic tumors with active neovascularization. HepG2 and Huh-6 xenografts showed expression of β-catenin, AFP, and Glypican-3 (GPC3). HepG2 samples displayed a consistent gene expression profile most similar to human HB tumors. Intrahepatic injection of HB cell lines leads to liver tumors in mice with growth patterns and biologic, histologic, and genetic features similar to human HB tumors. This orthotopic xenograft mouse model will enable clinically relevant testing of novel agents for HB.

Animal Modeling of Pediatric Liver Cancer

“…Figure 1 demonstrates this technique of intrahepatic injection of human tumor cells that was used in this study, with the resulting tumors shown. These intrahepatic HB xenograft models recapitulated the key hallmarks of the disease including elevated serum AFP levels, large exophytic tumors with active blood supplies, and embryonal histological phenotypes with elevation of AFP, Glypican-3 (GPC3), and β-catenin proteins [19]. These models, especially the HepG2 model, also displayed a consistent gene expression profile similar to human HB tumors [19].…”

“…These intrahepatic HB xenograft models recapitulated the key hallmarks of the disease including elevated serum AFP levels, large exophytic tumors with active blood supplies, and embryonal histological phenotypes with elevation of AFP, Glypican-3 (GPC3), and β-catenin proteins [19]. These models, especially the HepG2 model, also displayed a consistent gene expression profile similar to human HB tumors [19]. Noted in the study was the observation that gene expression of the Huh-6 cells was changed by growth in the murine liver and that this correlated with differences in histology [19].…”

“…Use of cell lines grown extensively in vitro [18,19] Subcutaneous PDX model Fresh patient samples more closely resemble primary disease Model does not accurately recapitulate tumor microenvironment and vascularization [20][21][22][23] Intrahepatic PDX model Fresh patient samples closely resemble primary disease and tumors recapitulate liver microenvironment Limited access to patient samples for model generation [24] LIN28B GEM model Use of immunocompetent animals and specific exploration of LIN28B…”

“…In 2017, Woodfield and colleagues developed several models with the growth of intrahepatic HB tumors from the commercially available HepG2 and Huh-6 cell lines [19]. In this work, cell lines were injected directly into either the right-median or left-lateral liver lobes of NOD/Shi-scid/IL-2Rγnull (NOG) mice [19]. Cells injected with both techniques grew large, extraphytic masses with local invasion indicative of PRETEXT 1, 2, and 3 disease.…”

Osthole attenuates angiogenesis in an orthotopic mouse model of hepatocellular carcinoma via the downregulation of nuclear factor‑κB and vascular endothelial growth factor

“…The orthotopic mouse model of HCC can be established by the injection of HCC cells or the tumor tissues into the mouse liver, and the growth of tumors can be monitored using imaging techniques (24,32,33). The present study directly injected the HCC Hepa1-6 cell line into the left liver lobes of mice to generate internal liver tumors, and Small Animal Diagnostic Ultrasound was performed at early (7 or 8 days) and late (20–27 days) time points to confirm the presence of a tumor and for the assessment of tumor burden, respectively.…”