Transposon-based oncogene integration in Abcb4(Mdr2)^-/- mice recapitulates high susceptibility to cholangiocarcinoma in primary sclerosing cholangitis

Pinzhu Huang¹, Guangyan Wei², Jesse D Kirkpatrick³, Yi Lin⁴, Li Tan⁵, Heansika Matta⁶, Imad Nasser⁷, Mingzhe Huang⁸, Li Chen⁹, Mathieu Petitjean⁹, Disha Skelton-Badlani⁶, Wen Gao⁶, Kahini Vaid⁶, Shuangshuang Zhao⁶, Alicia Lugovskoy⁶, Maram Alenzi⁶, Xin Chen¹⁰, Gregory J Gores¹¹, Yury V Popov¹²

Affiliations

¹ Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Department of Colon and Rectum Surgery, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
² Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Department of Radiation Oncology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
³ Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA; Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁴ Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Department of Gastroenterology and Hepatology, Fujian Provincial Hospital, Fuzhou, China.
⁵ Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Center of Hepato-Pancreato-Biliary Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
⁶ Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
⁷ Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
⁸ Department of Colon and Rectum Surgery, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
⁹ PharmaNest, Inc, NJ, USA.
¹⁰ University of Hawaii Cancer Center, Honolulu, HI USA.
¹¹ Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN, USA.
¹² Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA. Electronic address: ypopov@bidmc.harvard.edu.

PMID: 39089631
PMCID: PMC11655257 (available on 2026-01-01)
DOI: 10.1016/j.jhep.2024.07.016

Transposon-based oncogene integration in Abcb4(Mdr2)^-/- mice recapitulates high susceptibility to cholangiocarcinoma in primary sclerosing cholangitis

Pinzhu Huang et al. J Hepatol. 2025 Jan.

. 2025 Jan;82(1):84-96.

doi: 10.1016/j.jhep.2024.07.016. Epub 2024 Jul 30.

Authors

Affiliations

¹ Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Department of Colon and Rectum Surgery, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
² Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Department of Radiation Oncology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
³ Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA; Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁴ Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Department of Gastroenterology and Hepatology, Fujian Provincial Hospital, Fuzhou, China.
⁵ Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Center of Hepato-Pancreato-Biliary Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
⁶ Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
⁷ Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
⁸ Department of Colon and Rectum Surgery, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
⁹ PharmaNest, Inc, NJ, USA.
¹⁰ University of Hawaii Cancer Center, Honolulu, HI USA.
¹¹ Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN, USA.
¹² Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA. Electronic address: ypopov@bidmc.harvard.edu.

PMID: 39089631
PMCID: PMC11655257 (available on 2026-01-01)
DOI: 10.1016/j.jhep.2024.07.016

Abstract

Background & aims: Cholangiocarcinoma (CCA) is a dreaded complication of primary sclerosing cholangitis (PSC) that is difficult to diagnose and associated with high mortality. A lack of animal models of CCA recapitulating the hepatic microenvironment of sclerosing cholangitis has hindered the development of novel treatments. Herein, we sought to develop a mouse model of PSC-associated CCA.

Methods: Ten-week-old Mdr2^-/- mice with congenital PSC-like disease, and healthy wild-type littermates were subjected to either modified retrograde biliary instillation or hydrodynamic tail vein injection of a sleeping beauty transposon-transposase plasmid system with activated AKT (myr-AKT) and Yap (YapS127A) proto-oncogenes (SB AKT/YAP1). The role of TGFβ was interrogated via ALK5 inhibitor (SB-525334) administration. Tumor phenotype, burden and desmoplastic reaction were analyzed histologically and via RNA sequencing.

Results: While SB AKT/YAP1 plasmids administered via retrograde biliary injection caused tumors in Mdr2^-/-, only 26.67% (4/15) of these tumors were CCA. Alternatively, hydrodynamic tail vein injection of SB AKT/YAP1 resulted in robust tumorigenesis in all fibrotic Mdr2^-/- mice with high CCA burden compared to healthy mice. Tumors phenotypically resembled human CCA, expressed multiple CCA (but not hepatocellular carcinoma) markers, and exhibited a profound desmoplastic reaction. RNA sequencing analysis revealed profound transcriptional changes in CCA evolving in a PSC-like context, with specific alterations in multiple immune pathways. Pharmacological TGFβ inhibition led to enhanced immune cell tumor infiltration, reduced tumor burden and suppressed desmoplastic collagen accumulation compared to placebo.

Conclusion: We established a new high-fidelity cholangiocarcinoma model in mice, termed SB CCA.Mdr2^-/-, which recapitulates the increased susceptibility to CCA in the setting of biliary injury and fibrosis observed in PSC. Through transcriptomics and pharmacological studies, we show dysregulation of multiple immune pathways and TGFβ signaling as potential drivers of CCA in a PSC-like microenvironment.

Impact and implications: Animal models for primary sclerosing cholangitis (PSC)-related cholangiocarcinoma (PSC-CCA) are lacking. Thus, we have developed and characterized a new mouse model of PSC-CCA, termed SB CCA.Mdr2^-/-, which features reliable tumor induction on a PSC-like background of biliary injury and fibrosis. Global gene expression alterations were identified and standardized tools, including automated whole slide image analysis methodology for tumor burden and feature analysis, were established to enable systematic research into PSC-CCA biology and formal preclinical drug testing.

Keywords: ABCB4; TGFβ; cholangiocarcinoma; mouse model; primary sclerosing cholangitis.

PubMed Disclaimer

Conflict of interest statement

Conflict of interest statement: Li Chen and Mathieu Petitjean are employees of PharmaNest, Inc developing and marketing the software tools used for image analysis in this study. No other potential conflict of interest.

References

1. Banales JM, Marin JJG, Lamarca A, et al. Cholangiocarcinoma 2020: the next horizon in mechanisms and management. Nat Rev Gastroenterol Hepatol 2020;17:557–588. - PMC - PubMed
1. Razumilava N, Gores GJ. Cholangiocarcinoma. Lancet 2014;383:2168–2179. - PMC - PubMed
1. Sirica AE, Gores GJ, Groopman JD, et al. Intrahepatic Cholangiocarcinoma: Continuing Challenges and Translational Advances. Hepatology 2019;69:1803–1815. - PMC - PubMed
1. Hogdall D, Lewinska M, Andersen JB. Desmoplastic Tumor Microenvironment and Immunotherapy in Cholangiocarcinoma. Trends Cancer 2018;4:239–255. - PubMed
1. Patel N, Benipal B. Incidence of Cholangiocarcinoma in the USA from 2001 to 2015: A US Cancer Statistics Analysis of 50 States. Cureus 2019;11:e3962. - PMC - PubMed

MeSH terms

Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions

Substances

Actions
Actions
Actions
Actions
Actions
Actions

Grants and funding

R01 DK139288/DK/NIDDK NIH HHS/United States

LinkOut - more resources

Full Text Sources
- Elsevier Science

Save citation to file

Email citation

Add to Collections

Add to My Bibliography

Your saved search

Create a file for external citation management software

Your RSS Feed

Transposon-based oncogene integration in Abcb4(Mdr2)^-/- mice recapitulates high susceptibility to cholangiocarcinoma in primary sclerosing cholangitis

Affiliations

Transposon-based oncogene integration in Abcb4(Mdr2)^-/- mice recapitulates high susceptibility to cholangiocarcinoma in primary sclerosing cholangitis

Authors

Affiliations

Abstract

Conflict of interest statement

References

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources