Portal Vein Injection of Colorectal Cancer Organoids to Study the Liver Metastasis Stroma

Hiroki Kobayashi; Krystyna A. Gieniec; Jia Q. Ng; Jarrad Goyne; Tamsin R. M. Lannagan; Elaine M. Thomas; Georgette Radford; Tongtong Wang; Nobumi Suzuki; Mari Ichinose; Josephine A. Wright; Laura Vrbanac; Alastair D. Burt; Masahide Takahashi; Atsushi Enomoto; Daniel L. Worthley; Susan L. Woods

doi:10.3791/62630

Method Article

Portal Vein Injection of Colorectal Cancer Organoids to Study the Liver Metastasis Stroma

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DOI:

10.3791/62630

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September 3rd, 2021

Hiroki Kobayashi¹^,²^,³^,⁴ , Krystyna A. Gieniec¹^,² , Jia Q. Ng¹^,² , Jarrad Goyne¹^,² , Tamsin R. M. Lannagan¹^,² , Elaine M. Thomas¹^,² , Georgette Radford¹^,² , Tongtong Wang¹^,² , Nobumi Suzuki¹^,²^,⁵ , Mari Ichinose¹^,² , Josephine A. Wright² , Laura Vrbanac¹^,² , Alastair D. Burt⁶ , Masahide Takahashi³^,⁴^,⁷ , Atsushi Enomoto³ , Daniel L. Worthley² , Susan L. Woods¹^,²

¹Adelaide Medical School, University of Adelaide, ²South Australian Health and Medical Research Institute (SAHMRI), ³Department of Pathology, Nagoya University Graduate School of Medicine, ⁴Division of Molecular Pathology, Center for Neurological Disease and Cancer, Nagoya University Graduate School of Medicine, ⁵Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, ⁶Translational and Clinical Research Institute, Newcastle University, ⁷International Center for Cell and Gene Therapy, Fujita Health University

Summary

Portal vein injection of colorectal cancer (CRC) organoids generates stroma-rich liver metastasis. This mouse model of CRC hepatic metastasis represents a useful tool to study tumor-stroma interactions and develop novel stroma-directed therapeutics such as adeno-associated virus-mediated gene therapies.

Abstract

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the tumor microenvironment, play a crucial role in metastatic CRC progression and predict poor patient prognosis. However, there is a lack of satisfactory mouse models to study the crosstalk between metastatic cancer cells and CAFs. Here, we present a method to investigate how liver metastasis progression is regulated by the metastatic niche and possibly could be restrained by stroma-directed therapy. Portal vein injection of CRC organoids generated a desmoplastic reaction, which faithfully recapitulated the fibroblast-rich histology of human CRC liver metastases. This model was tissue-specific with a higher tumor burden in the liver when compared to an intra-splenic injection model, simplifying mouse survival analyses. By injecting luciferase-expressing tumor organoids, tumor growth kinetics could be monitored by in vivo imaging. Moreover, this preclinical model provides a useful platform to assess the efficacy of therapeutics targeting the tumor mesenchyme. We describe methods to examine whether adeno-associated virus-mediated delivery of a tumor-inhibiting stromal gene to hepatocytes could remodel the tumor microenvironment and improve mouse survival. This approach enables the development and assessment of novel therapeutic strategies to inhibit hepatic metastasis of CRC.

Introduction

Colorectal cancer (CRC) is a major cause of cancer mortality worldwide¹. More than half of the CRC patients develop hepatic metastasis that occurs through the portal vein dissemination¹. Currently, there are no effective therapeutics that can cure advanced liver metastasis, and most patients succumb to metastatic disease.

The metastatic niche or tumor microenvironment plays a key role in engraftment and growth of disseminated CRC cells². Cancer-associated fibroblasts (CAFs), a prominent component of the tumor microenvironment, promote or restrain cancer progression thro....