Patient-derived tumor organoid and fibroblast assembloid models for interrogation of the tumor microenvironment in esophageal adenocarcinoma

Benjamin P Sharpe; Liliya A Nazlamova; Carmen Tse; David A Johnston; Jaya Thomas; Rhianna Blyth; Oliver J Pickering; Ben Grace; Jack Harrington; Rushda Rajak; Matthew Rose-Zerilli; Zoe S Walters; Tim J Underwood

doi:10.1016/j.crmeth.2024.100909

Patient-derived tumor organoid and fibroblast assembloid models for interrogation of the tumor microenvironment in esophageal adenocarcinoma

Cell Rep Methods. 2024 Nov 20:100909. doi: 10.1016/j.crmeth.2024.100909. Online ahead of print.

Authors

Affiliations

¹ School of Cancer Sciences, Faculty of Medicine, University of Southampton, Southampton SO16 6YD, UK.
² School of Cancer Sciences, Faculty of Medicine, University of Southampton, Southampton SO16 6YD, UK; Early Cancer Institute, University of Cambridge, Department of Oncology, Box 197 Cambridge Biomedical Campus, Hills Road, Cambridge SB2 0XZ, UK.
³ Biomedical Imaging Unit, Faculty of Medicine, University of Southampton, Southampton SO16 6YD, UK.
⁴ Bio-R Bioinformatics Research Facility, Faculty of Medicine, University of Southampton, Southampton SO16 6YD, UK.
⁵ School of Cancer Sciences, Faculty of Medicine, University of Southampton, Southampton SO16 6YD, UK; University Hospital Southampton NHS Foundation Trust, Southampton SO16 6YD, UK.
⁶ School of Cancer Sciences, Faculty of Medicine, University of Southampton, Southampton SO16 6YD, UK; Southampton Clinical Trials Unit, University of Southampton, Southampton, UK.
⁷ University Hospital Southampton NHS Foundation Trust, Southampton SO16 6YD, UK.
⁸ School of Cancer Sciences, Faculty of Medicine, University of Southampton, Southampton SO16 6YD, UK; Bio-R Bioinformatics Research Facility, Faculty of Medicine, University of Southampton, Southampton SO16 6YD, UK. Electronic address: tju@soton.ac.uk.

PMID: 39610249
DOI: 10.1016/j.crmeth.2024.100909

Abstract

The tumor microenvironment (TME) comprises all non-tumor elements of cancer and strongly influences disease progression and phenotype. To understand tumor biology and accurately test new therapeutic strategies, representative models should contain both tumor cells and normal cells of the TME. Here, we describe and characterize co-culture tumor-derived organoids and cancer-associated fibroblasts (CAFs), a major component of the TME, in matrix-embedded assembloid models of esophageal adenocarcinoma (EAC). We demonstrate that the assembloid models faithfully recapitulate the differentiation status of EAC and different CAF phenotypes found in the EAC patient TME. We evaluate cell phenotypes by combining tissue-clearing techniques with whole-mount immunofluorescence and histology, providing a practical framework for the characterization of cancer assembloids.

Keywords: CP: Cancer biology; CP: Stem cell; assembloids; cancer-associated fibroblasts; esophageal adenocarcinoma; organoids; tumor microenvironment.