Periodic Reporting for period 4 - COMIET (Engineering Complex Intestinal Epithelial Tissue Models)
Período documentado: 2020-06-01 hasta 2021-05-31
2. Today, advancing a drug candidate to regulatory approval takes roughly 12 to 15 years and costs an estimated $800 to $900 million on average. As over 50% of drug candidates fail early, discarding of compounds in the in vitro preclinical tests saves costs, time and reduces the ethical concerns of indiscriminate usage of animal testing. On the other hand, the benefits of having an advanced model of gut to mimic digestive diseases and disorders is huge. The quality of life of people affected with inflammatory bowel disease is significantly lower than in healthy persons. Since therapeutic approaches are only symptomatic, the development of in vitro human intestinal models might contribute to shed light on the basic biology of these diseases by proposing new therapeutic strategies.
3. In this context, COMIET has made use of microengineering technologies and adult stem cells to develop artificial intestinal tissues that mimic the physiological characteristics found in vivo to open new areas of research on human intestinal diseases. The novel in vitro models of intestinal epithelial tissues aim to address intestinal biology such as cell renewal and migration, multicell-type differentiation, and cell-cell interactions to go beyond the state-of-the-art 3D minigut (organoid) in vitro models. The cell culture platforms proposed aimed to provide physiologically relevant and highly reproducible data, being compatible with conventional cell culture assays and high-throughput testing. To achieve this, we have developed an experimental setup that combines microfabrication technologies, tissue engineering and advances in intestinal stem cell research, exploiting stem cell self-organizing characteristics.
2. We have demonstrated that intestinal cells cultured on our substrates behave in a more physiological manner than cells grown on conventional substrates. They better recreate the intestinal barrier properties: adherence and invasion capacity of E. coli LF82 pathogenic bacterial (García-Diaz et al., CRS Conference 2019), permeability and drug absorption properties (Castaño et al., Biofabrication 2019) and allow mimicking the interaction between different cell types of the intestinal mucosa (Vila et al., Biofabrication 2020), which is crucial to disease modelling (A. Vila, PhD thesis 2020). To better exploit this result we have established an on-going collaboration with a local hospital (Hospital Vall d’Hebron).
3. We have successfully developed and characterized several hydrogel formulations that can be used as bioinks in light-based applications (Castaño et al., Biofabrication 2019; Vila et al., Biofabrication 2020; Borgheti et al., in preparation). Aside of the scientific dissemination, we have established an agreement with a Spanish company that develops bioprinting instruments (Regemat 3D) to transfer a bioink specific formulation to their product catalogue.
4. Organoid-derived intestinal epithelial cells can grow on our 3D hydrogels and, upon delivering specific molecular gradients through a microfluidic device, cells are compartmentalized thus recreating their position in the in vivo tissue (G. Altay, PhD thesis 2019; Altay et al., 10.1101/2021.12.13.472418; Martinez et al, Biomaterials Conf. 2020; Martinez et al., MRS fall conf. 2020). We have established a protocol to fabricate the scaffold, the device and to “open-up” intestinal organoids onto monolayers (Altay et al. Scie Reports 2019; Altay et al., Bio-protocol 2020).
5. We successfully developed 2D approaches that have proven to be useful technological tools for the study of relevant factors recognized to affect intestinal cell behavior such as the stiffness of the substrate (Comelles et al., Biofabrication 2020), the patterning of relevant biomolecules identified in the intestinal development and homeostasis (Hortigüela et al., Nano Letters 2018; Cutrale et al., Nat. Protocols 2019; E. Larrañaga, PhD thesis 2021) and new 3D set-ups to study epithelial-stromal interactions (Fernández-Majada et al., 10.1101/2021.05.28.446131).