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

Increased extracellular matrix (ECM) stiffness has been implicated in esophageal adenocarcinoma (EAC) progression, metastasis, and resistance to therapy. However, the underlying pro-tumorigenic 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 to identify potential therapeutic targets to disrupt the contribution of pro-tumorigenic 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. Overall design: Samples are patient-derived 3D esophageal adenocarcinoma (EAC) organoids grown in 4 different hydrogel conditions. Only one organoid line was used. Each hydrogel condition was performed in triplicates, for a total of 12 samples. Samples 4, 8 and 12 are the reference group, as triplicates, which are EAC organoids grown in Matrigel of 100 Pa of storage modulus. Samples 3, 7 and 11 are EAC organoids grown in NorHA hydrogels of 100 Pa of storage modulus, in triplicates. Samples 2, 6 and 10 are EAC organoids grown in NorHA hydrogels of 350 Pa of storage modulus, in triplicates. Samples 1, 5 and 9 are EAC organoids grown in NorHA hydrogels of 1000 Pa of storage modulus, in triplicates.