Patient-Derived Scaffolds of Colorectal Cancer Metastases as an Organotypic 3D Model of the Liver Metastatic Microenvironment

Abstract

The liver is the most common site for colorectal cancer (CRC) metastasis and there is an urgent need for new tissue culture models to study colorectal cancer liver metastasis (CRLM) as current models do not mimic the biological, biochemical, and structural characteristics of the metastatic microenvironment. Decellularization provides a novel approach for the study of the cancer extracellular matrix (ECM) as decellularized scaffolds retain tissue-specific features and biological properties. In the present study, we created a 3D model of CRC and matched CRLM using patient-derived decellularized ECM scaffolds seeded with the HT-29 CRC cell line. Here, we show an increased HT-29 cell proliferation and migration capability when cultured in cancer-derived scaffolds compared to same-patient healthy colon and liver tissues. HT-29 cells cultured in CRLM scaffolds also displayed an indication of epithelial-mesenchymal transition (EMT), with a loss of E-cadherin and increased Vimentin expression. EMT was confirmed by gene expression profiling, with the most represented biological processes in CRLM-seeded scaffolds involving demethylation, deacetylation, a cellular response to stress metabolic processes, and a response to the oxygen level and starvation. HT-29 cells cultured in cancer-specific 3D microenvironments showed a reduced response to treatment with 5-fluorouracil and 5-fluorouracil combined with Irinotecan when used at a standard IC50 (as determined in the 2D culture). Our 3D culture system with patient-derived tissue-specific decellularized ECM better recapitulates the metastatic microenvironment compared to conventional 2D culture conditions and represents a relevant approach for the study of CRLM progression and assessing the response to chemotherapy agents.

Immunomodulatory Role of the Extracellular Matrix Within the Liver Disease Microenvironment

“…Decellularization describes the treatment of a tissue or whole organ with a series of enzymes and detergents to effectively remove the native cellular content and DNA, while maintaining the ECM network. Decellularized scaffolds are tissue-and function-specific based on their origin and can be combined with different cell populations to create 3D models (423)(424)(425).…”

3D Culture Systems for Exploring Cancer Immunology

“…They found that HT-29 cells grown in cancer-derived matrix had increased proliferation, migration and reduced sensitivity to common chemotherapies. HT-29 cells grown in metastases-derived scaffolds had increased epithelial-to-mesenchymal transition phenotype [ 105 ]. With respect to cancer immunology, Pinto et al demonstrated that, unlike healthy colon matrix, colorectal cancer-derived matrix polarized macrophages to the anti-inflammatory M2 phenotype, enhancing macrophage production of immunosuppressive cytokines [ 106 ].…”

Recellularized Colorectal Cancer Patient-Derived Scaffolds as In Vitro Pre-Clinical 3D Model for Drug Screening

“…In this novel in vitro CRC model, we evaluated the activity of specific drugs with the aim to translate it as a preclinical model before chemotherapy administration in patients. We recently demonstrated that the decellularized healthy tissues and neoplastic counterparts were biologically active scaffolds able to support the in vitro cell growth and proliferation [14,18]. The decellularization process enables complete cell removal, maintaining at the same time the tissue architecture and preserving the ECM components [19].…”