Spatiotemporally resolved ex vivo colorectal cancer development in engineered mini-colons (scRNA-Seq)
Ontology highlight
ABSTRACT: Here we developed topobiologically complex mini-colons able to undergo tumorigenesis ex vivo by integrating microfabrication, optogenetic, and tissue engineering approaches. With this system, tumorigenic transformation can be spatiotemporally controlled by directing oncogenic activation through blue-light exposure, and emerging colon tumors can be tracked in real-time with single-cell resolution for several weeks without breaking the culture. These induced mini-colons display rich intra- and inter-tumoral diversity and recapitulate key pathophysiological hallmarks displayed by colorectal tumors in vivo. By fine-tuning cell-intrinsic and extrinsic parameters, mini-colons can be leveraged to unveil tumorigenic determinants, including dietary patterns, microbiota-derived metabolites, and pharmacological therapies.
Project description:Here we developed topobiologically complex mini-colons able to undergo tumorigenesis ex vivo by integrating microfabrication, optogenetic, and tissue engineering approaches. With this system, tumorigenic transformation can be spatiotemporally controlled by directing oncogenic activation through blue-light exposure, and emerging colon tumors can be tracked in real-time with single-cell resolution for several weeks without breaking the culture. These induced mini-colons display rich intra- and inter-tumoral diversity and recapitulate key pathophysiological hallmarks displayed by colorectal tumors in vivo. By fine-tuning cell-intrinsic and extrinsic parameters, mini-colons can be leveraged to unveil tumorigenic determinants, including dietary patterns, microbiota-derived metabolites, and pharmacological therapies.
Project description:Here we developed topobiologically complex mini-colons able to undergo tumorigenesis ex vivo by integrating microfabrication, optogenetic, and tissue engineering approaches. With this system, tumorigenic transformation can be spatiotemporally controlled by directing oncogenic activation through blue-light exposure, and emerging colon tumors can be tracked in real-time with single-cell resolution for several weeks without breaking the culture. These induced mini-colons display rich intra- and inter-tumoral diversity and recapitulate key pathophysiological hallmarks displayed by colorectal tumors in vivo. By fine-tuning cell-intrinsic and extrinsic parameters, mini-colons can be leveraged to unveil tumorigenic determinants, including dietary patterns, microbiota-derived metabolites, and pharmacological therapies.
Project description:The compromise to reduce, replace, and refine (3Rs) animal usage in cancer research demands the development of biologically capable ex vivo cancer models. The establishment of three-dimensional organoid-based systems has paved the way in this direction by bringing some realistic in vivo features to the in vitro context. However, faithful modeling of cancer requires a degree of tissue-level organization, multi-cellular diversity, biological durability, and experimental flexibility that far exceeds the capabilities of any existing in vitro systems. Here we implemented tissue engineering and microfabrication technologies to develop topobiologically complex patient-specific avatars of colorectal cancer able to overcome those limitations. These miniature tissues consist of long-lived gut-like-shaped healthy colon epithelia (“mini-colons”) that allow the stable integration of cancer cells and their native tumor microenvironment on a platform designed for the real-time high-resolution evaluation of cellular dynamics. This provides an unprecedented repertoire of ex vivo experimental possibilities, which we illustrate through different applications, including the discovery of a cancer associated fibroblast-triggered mechanism driving colorectal cancer invasion. As a whole, our mini-colon system pushes the boundaries of ex vivo cancer research in both basic and pre-clinical settings.