ProTeAN · Production and Testing of humAn-derived Neurons and brain organoids: advanced model probing in neurodevelopmental disorders

Production and Testing of humAn-derived Neurons and brain organoids: advanced model probing in neurodevelopmental disorders

The ProTeAN project was an innovative and ambitious project that combined expertise from academic and clinical partners to develop 3D models to study disorders that arise during brain development, at the Center for Neuroscience and Cell Biology, Portugal. Autism spectrum disorder (ASD), intellectual disability and attention-deficit/hyperactivity disorder, affect over 3% of children worldwide. ASD alone is estimated to afflict 1 in 150 children and its annual cost to society has been proposed to run as high as 400 billion euros in the EU. Clinical trials to test drugs for autism have repeatedly failed as previous studies using mouse models may not accurately replicate what occurs in the human body. Therefore, it is critical to develop realistic models that will advance knowledge on brain-related disorders and to test therapeutic strategies. The goal of the ProTeAN project was to implement the required methodologies of developing 3D models of brain development, named “brain organoids”. This allows the neurons, the building blocks of the brain, to grow in a three dimensional environment and form complex structures as occurs in humans, instead of growing these cells on a plastic surface, that could have been hiding the actual biological processes that are affected during brain development. To generate the brain organoids, we collected tissue samples from deciduous teeth or third molars from ASD patients and controls, through a minimally invasive procedure, in comparison to other more invasive procedures, such as skin biopsies. By using samples directly from patients, we will be able to later test therapeutic strategies, in a personalised medicine approach. The first milestones we achieved were the establishment of the first dental stem cell biobank in Portugal to study brain disorders and its approval by the Ethics Committees from the Pediatric Hospital of Coimbra and the University of Coimbra. We were able to optimize the required protocols to transform the dental stem cells into induced pluripotent stem cells that can become virtually any cell type of the body. Therefore, we used the induced pluripotent stem cells to obtain neurons arranged in a 3D conformation that would then become the more organized brain organoids. Brain organoids were then characterized to analyze any alterations in the morphology of their neurons and in the connectivity between them. We showed that dental stem cells are a suitable source to generate brain organoids and that these 3D brain models are realistic models that will allow us to understand the mechanisms behind brain disorders and test for personalized therapeutic approaches. Regarding science communication and outreach, this project was widely disseminated through national and international media and also through outreach activities to raise autism awareness and to educate the community on the scientific advancements achieved in this research field. In conclusion, the ProTeAN project represented a novel line of research at CNC, the host institution, and it was crucial for the implementation and optimization all the methodologies required for brain organoid development and characterization. We believe that now we are fully equipped to advance this project at a fast pace and be able to perform comparisons between brain organoids from patients and controls, and ultimately identify therapeutic strategies in a personalised medicine approach.

Data: CORDIS, © European Union