Engineered hydrogel reveals contribution of matrix mechanics to esophageal adenocarcinoma and identifies matrix-activated therapeutic targets

Abstract

Increased extracellular matrix (ECM) stiffness has been implicated in esophageal adenocarcinoma (EAC) progression, metastasis, and resistance to therapy. However, the underlying protumorigenic pathways are yet to be defined. Additional work is needed to develop physiologically relevant in vitro 3D culture models that better recapitulate the human tumor microenvironment and can be used to dissect the contributions of matrix stiffness to EAC pathogenesis. Here, we describe a modular, tumor ECM–mimetic hydrogel platform with tunable mechanical properties, defined presentation of cell-adhesive ligands, and protease-dependent degradation that supports robust in vitro growth and expansion of patient-derived EAC 3D organoids (EAC PDOs). Hydrogel mechanical properties control EAC PDO formation, growth, proliferation, and activation of tumor-associated pathways that elicit stem-like properties in the cancer cells, as highlighted through in vitro and in vivo environments. We also demonstrate that the engineered hydrogel serves as a platform for identifying potential therapeutic targets to disrupt the contribution of protumorigenic matrix mechanics in EAC. Together, these studies show that an engineered PDO culture platform can be used to elucidate underlying matrix-mediated mechanisms of EAC and inform the development of therapeutics that target ECM stiffness in EAC.

Modelling esophageal adenocarcinoma and Barrett’s esophagus with patient-derived organoids

“…While our primary emphasis in this discussion has been on the composition of the culture media as a key factor influencing the rate of organoid establishment, it is imperative to recognize that other variables, including the quality of sampling, sample digestion, matrix scaffold, and technical handling, also play a vital role. Although most of these factors have not been examined concerning their effects on EAC PDO culture, a recent study attempted to explore how altering mechanical and biochemical matrix properties could impact growth and mutational profile of EAC PDOs ( Cruz-Acuña et al, 2023 ). Initially, this study demonstrated that varying the concentration of Matrigel resulted in changes in the growth and transcriptional expression of EAC-associated genes (e.g., TP53 , NOTCH1 and ZEB1 ).…”

“…Matrigel batches exhibit variability in these properties, making it difficult to recapitulate the independent effects of each. To bridge this knowledge gap, Cruz-Acuña et al, 2023 designed a hydrogel matrix to mimic the extracellular matrix (ECM) found in EAC tumors. This matrix is constructed using hyaluronic acid (HA) and is specifically crafted to include adhesive peptides and protease-degradable peptide crosslinkers.…”

Organoids in gastrointestinal diseases: from bench to clinic

“… 180 , 181 They also serve as a means to continuously identify new targets and design targeted therapy strategies. 182 , 183 , 184 , 185 Organoids models also facilitate the exploration of alternative treatment approaches for challenging drug targets. For instance, though certain therapeutic strategies targeting the WNT pathway often lead to adverse effects on normal crypt development, innovative approaches focusing on terminal differentiation have shown promise in resolving these issues.…”

“…Furthermore, organoids models are employed for evaluating repurposed drugs originally intended for other diseases and assessing their efficacy against novel diseases 180,181 . They also serve as a means to continuously identify new targets and design targeted therapy strategies 182–185 . Organoids models also facilitate the exploration of alternative treatment approaches for challenging drug targets.…”