Patient-derived disease modeling as a tool to uncover the molecular pathogenesis of neurodevelopmental disorders (NDDs)

Title Patient-derived disease modeling as a tool to uncover the molecular pathogenesis of neurodevelopmental disorders (NDDs)
Acronym NDD, organoids, disease modelling
Start date 2018-10-01
End date 2022-09-30
Sponsor Marie Skłodowska-Curie Action (MSCA)
Institution University of Milan

Project Description

Being multifaceted conditions characterized by impairments in cognition, communication, behavior and motor skills, neurodevelopmental disorders (NDDs) result from abnormal CNS development. Our poor understanding of the molecular pathogenesis of NDDs led researchers to look for alternative methodologies to study the mechanisms underlying these disorders. A breakthrough in human disease research over the last decade has emerged from cellular reprogramming technologies, using reprogrammed patient somatic cells because it captures a patient's genome in a pluripotent stage allowing us to study, for the first time, the initial development and progression of pathology in live human cells in a controlled environment. Here we aim to dissect the molecular pathogenesis of a group of NDDs using patient-specific induced pluripotent stem cells (iPSCs) generated from skin biopsies of affected patients and matched unaffected relatives as well as 3D cortical organoids to functionally dissect the impact and variability of these disorders. We will use a large cohort of iPSC lines from two different paradigmatic neurodevelopmental disorders such as Weaver Syndrome (WS) and Kabuki Syndrome (KS) caused by mutations, respectively, in EZH2 (catalysing H2K27 tri-methylation) and KMT2D or KDM6A (catalysing H3K4 mono-methylation and di- and trimethyl H3K27 demethylation, respectively), as well as Gabriele-DeVries syndrome (GADVES) characterized by mutations in YY1 (mediates gene activation by looping enhancer and promoter regions). After accessing major dysregulations in patients in both primary fibroblasts and iPSCs, we will move on to disease-relevant cell types such as neural crest stem cells (NCSCs) and cortical excitatory neurons representative of cortical layer IV since a variable intellectual disability (ID) is present in the above-mentioned syndromes. After differentiation into cortical neurons from patients and half-matched controls and following RNA-seq analysis of these cells, we will evaluate the possible dysregulations at the transcriptional level. To sustain the above-mentioned, we will perform an in-depth characterization of these deficits in terminal differentiation using functional and morphological tools, namely extracellularly recording the spontaneous neuronal network activity and measure morphometric properties using 2D and 3D preparations. By using disease-modeling as a tool to investigate the pathogenesis of NDDs across different time points, tissues and preparations will allow us to gather new insights about the molecular pathogenesis of these disorders.