Journal Article

Genetically engineered human cortical spheroid models of tuberous sclerosis

Bateup, Helen S., Blair, John D., Hockemeyer, Dirk

2018

Overview

Tuberous sclerosis complex (TSC) is a multisystem developmental disorder caused by mutations in the TSC1 or TSC2 genes, whose protein products are negative regulators of mechanistic target of rapamycin complex 1 signaling. Hallmark pathologies of TSC are cortical tubers—regions of dysmorphic, disorganized neurons and glia in the cortex that are linked to epileptogenesis. To determine the developmental origin of tuber cells, we established human cellular models of TSC by CRISPR–Cas9-mediated gene editing of TSC1 or TSC2 in human pluripotent stem cells (hPSCs). Using heterozygous TSC2 hPSCs with a conditional mutation in the functional allele, we show that mosaic biallelic inactivation during neural progenitor expansion is necessary for the formation of dysplastic cells and increased glia production in three-dimensional cortical spheroids. Our findings provide support for the second-hit model of cortical tuber formation and suggest that variable developmental timing of somatic mutations could contribute to the heterogeneity in the neurological presentation of TSC. CRISPR–Cas9-mediated gene editing of TSC1 and TSC2 in human pluripotent stem cells is used to investigate the contribution of tuberous sclerosis complex–mechanistic target of rapamycin complex 1 signaling to human neural development in two-dimensional monolayer and three-dimensional spheroid models of the neurodevelopmental disorder tuberous sclerosis complex.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 631/378/1689/2608 / 631/532 / 631/80/304 / 692/699/375/366 / Biomedical and Life Sciences / Biomedicine / Brain / Cancer Research / Care and treatment / Cell Line / Cerebral Cortex - metabolism / Cerebral Cortex - pathology / Cortex / CRISPR / CRISPR-Cas Systems - genetics / Deactivation / Development and progression / Developmental disabilities / Epilepsy / Gene editing / Gene mutation / Genes / Genetic Engineering / Genetic modification / Genetically modified organisms / Genome editing / Heterogeneity / Humans / Inactivation / Infectious Diseases / Metabolic Diseases / Molecular Medicine / Mutation / Neuroglia - metabolism / Neuroglia - pathology / Neuronal-glial interactions / Neurons / Neurons - metabolism / Neurons - pathology / Neurosciences / Pluripotency / Pluripotent Stem Cells - transplantation / Proteins / Rapamycin / Regulators / Spheroids / Spheroids, Cellular - metabolism / Stem cells / Three dimensional models / TOR protein / Tuberous sclerosis / Tuberous Sclerosis - genetics / Tuberous Sclerosis - physiopathology / Tuberous Sclerosis Complex 1 / Tuberous Sclerosis Complex 1 Protein - genetics / Tuberous Sclerosis Complex 2 / Tuberous Sclerosis Complex 2 Protein - genetics