Application of a 3D model for studying cortical neurogenesis and the WNT pathway using hiPSCs from a patient with autism spectrum disorder and macrocephaly

Abstract

The conventional 2D cell culture system has been used for studies of various aspects of the Central Nervous System. However, this type of culture does not reproduce the three-dimensional (3D) framework in which cells are found in native tissue. In this context, 3D bioprinting technology overcomes the limitations of the 2D system, providing an environment closer to the physiological one. Several cell types have already been bioprinted in 3D, including neuroprogenitor cells derived from human induced pluripotent stem cells (hiPSC-NPCs). Our laboratory has developed a new 3D model, produced by bioprinting hiPSC-NPCs, for studies of the main biological events of neurogenesis: proliferation, migration, and differentiation. This platform that we have developed has the potential for modeling not only physiological neurogenesis but also neurodevelopmental disorders, such as Autism Spectrum Disorder (ASD) and macrocephaly. The WNT signaling pathway is one of the pathways that orchestrates embryonic neurogenesis and has been described as dysregulated in patients diagnosed with ASD and macrocephaly. Thus, in this project, we propose to use the neurogenic niche model produced by 3D bioprinting of hiPSC-NPCs derived from a patient with ASD and macrocephaly and normal hiPSC-NPCs to study cortical neurogenesis and the WNT pathway in neurodevelopmental pathologies. The cytoarchitecture of the bioprinted tissue will be characterized, and interventions in the WNT pathway will be performed using agonists and antagonists to verify the contribution of this signaling pathway to the neurobiology of ASD and macrocephaly. We will use immunofluorescence followed by confocal microscopy, flow cytometry, qPCR, and western blot to identify alterations in the biological processes of neurogenesis that lead to the development of ASD.