Project Portugal 2030

Summary

CELLS2MODEL addresses an important issue regarding the development of a robust model to understand and study brain permeability to therapeutical drugs. In fact, the biggest challenge regarding the development of drugs targeting glioblastoma (GBM) and studying its pathophysiology is the presence of a blood-brain barrier (BBB). There have been significant advances in the development of in vitro BBB models due to the increasing awareness of the role of this barrier in health and diseases. Functional in vitro BBB models that mimic the in vivo environment provide a tool for drug screening and permeability studies and thus are of great pharmaceutical and clinical interest. Such models can provide a simplified biological system to study molecular mechanisms that are difficult to study in vivo. Currently, the most used in vitro BBB models rely on transwell filters with cultured endothelial brain cells (monoculture) or coculture with other cells from the NVU (astrocytes and pericytes), which are far from mimicking the complex in vivo scenario. To mitigate the lack of a robust multicellular model, CELLS2MODEL will employ cell-sheet engineering (CSE) to develop a 3D model that better mimics the native BBB (1st objective) not only in its cell composition (endothelial cells, pericytes, and astrocytes) but also in terms of extracellular matrix and basement membrane structure. The use of CSE is one of the novelties of CELLS2MODEL and will be backed by the PI's extensive experience in cell culture and background in tissue engineering strategies, as well as the long collaboration the host has with Okano's lab, a renowned pioneer in CSE and also by the host experience in this technique[23-26], that will provide a valuable input regarding scientific and technical aspects of this technology, as well as regarding its characterization. CSE has been applied to the regeneration of different tissues but its application for the development of a BBB model has never been reported and thus, this is an original concept and an innovative approach. CSE was selected to obtain the 3D multicellular model of the BBB due to its advantages in maintaining the extracellular matrix of each cell type and maintaining the cell-cell interactions intact. To better mimic the in vivo microenvironment, the assembled model will be further studied using a specifically designed microfluidic device to allow the evaluation of the model barrier function. The other breakthrough of CELL2MODEL is the combination of a 3D GBM model with the BBB to obtain a more complex system that better mimics the in vivo scenario. This will be possible using the designed microfluidic device. Validation is an essential component to complete the project. Therefore, we will use drugs that are currently used to treat GBM and the molecules that we have previously developed (Fmoc-Glucosamine-6-Phosphate, Fmoc-Glc6P) and tested for GBM cells to validate the model in vitro [8, 12, 28, 29]. The 3D GBM-BBB model that we will create will operate as a prototype for demonstration purposes. It can be tailored to facilitate personalized medicine by swapping out the GBM spheroids with patient-derived GBM organoids, which can then be incorporated into the BBB model. These bespoke models can be used for personalized drug screening.