Building human spinal cord organoids (huspineorg) biomodels to study neural tube defects
- Autores: José Manuel Blanco
- Directores de la Tesis: Elisa Martí Gorostiza (dir. tes.), Florenci Serras Rigalt (tut. tes.)
- Lectura: En la Universitat de Barcelona ( España ) en 2023
- Idioma: inglés
- Tribunal Calificador de la Tesis: Cristina Pujades Corbi (presid.), Esteban Kleinman Hoijman (secret.), Leonardo Beccari (voc.)
- Programa de doctorado: Programa de Doctorado en Genética por la Universidad de Barcelona
- Materias:
- Enlaces
- Tesis en acceso abierto en: TESEO
- Resumen
The Central nervous system (CNS) originates from the coordinated events that result into neural specification and the morphogenic events that shape the neural tube (NT). Then, the embryonic NT should grow and generate all the cell diversity present in the healthy organ. The morphogenic events that shape the NT occur in two consecutive, radically different processes referred as primary and secondary neurulation, that can be followed at different anterio-posterior levels in the Spinal cord (SpC). The posterior NT is formed by secondary neurulation, in a process concomitant to body axis elongation and mediated by the specification of nueromesodermal progenitors (NMPs). Defects in this process lead to caudal neural tube defects (NTDs). In the first chapter of this thesis, a new human 3D in vitro model for posterior SpC is set up. In these human organoid models, the neural specification, the morphogenesis of the NT, and its grow can be followed. Here, it is characterized a human organoid model were human embryonic stem cells (hESC) are guided into NMPs, expressing SOX2 and BRA and then into neural progenitor cells (NPCs), which maintain the SOX2 and lose BRA expression as it happens in vivo. Moreover, the NPCs are organized as an epithelium surrounding a central lumen. NPCs locate the centrosome and cilia at the lumen surface, where the polarity complexes are organized mimicking the polarity features of NPCs characterized in vivo. Additionally, in parallel to this epithelialization, the cell rearranges that shape the hollow NT formation in vivo, like the cell intercalation driving lumen resolution, can be followed in this organoid model. In the second chapter of this thesis, a screening to identify new mature centrosome com- ponents that would be potentially control the NPCs proliferation/differentiation rates was done independently with an in vivo, using the chick embryo as model, and an in silico approach. Unfortunately, the in vivo approach faced technical limitations intrinsic of the chick embryo model that prevent any successful identification of new candidates. However, the in silico approach provided a list of candidates to start their functional analysis.
