Periodic Reporting for period 1 - MECHANOIDS (Probing and controlling the three-dimensional organoid mechanobiology)
Período documentado: 2019-09-01 hasta 2021-08-31
This project aimed to quantify and manipulate the mechanobiology of normal gut and colorectal cancer organoids, and other 3D cell tissues. We developed tools to quantify the 3D stresses applied by an organoid embedded in a gel. We also measured the tension, pressure and contractility exerted by cell tissues surrounding a “tumor-like surrogate” gel, key parameters in organ growth and tumour progression.
Pressure and contractility were related with key biological parameters such as geometry, cell adhesion and proliferation. Proteins were fluorescently tagged in the cells that allowed us to track individual cells, as well as monitoring their polarization and adhesion.
The mechanical state of the organoids was also modified through optogenetic means. Proteins were expressed in the cells to control Rho-GTPases upon illumination, thereby steering the local stresses at will. This allowed to modify organoid pressure, contractility and geometry.
As organoids emerge as novel tools in biomedical research, we expect that studying and manipulating their mechanobiology will bring key insight into disease and development processes, and potential new therapeutic targets.
The researcher created Colorectal Cancer organoid mutants expressing fluorescent proteins such as actin, CAAX and e-cadherin, and optogenetic contractility constructs through lentiviral transfection. These mutants and the previously mentioned tools were used to study the interplay between structure, shape and stresses.
The tools generated by the researcher were applied to the study of the mechanics of crypts in intestinal organoids, published in a peer-reviewed paper [https://doi.org/10.1038/s41556-021-00699-6]; and to study the interplay between Cancer-Associated Fibroblast contractility and tumor mechanotransduction, disseminated in the form of a preprint [https://doi.org/10.1101/2021.04.05.438443].