Abstract
Animal studies have indicated that SOX10 is one of the key transcription factors regulating the proliferation, migration and differentiation of multipotent neural crest (NC), and mutation of SOX10 in humans may lead to type 4 Waardenburg syndrome (WS). However, the exact role of SOX10 in human NC development and the underlying molecular mechanisms of SOX10-related human diseases remain poorly understood due to the lack of appropriate human model systems. In this study, we successfully generated SOX10-knockout human induced pluripotent stem cells (SOX10−/− hiPSCs) by the CRISPR-Cas9 gene editing tool. We found that loss of SOX10 significantly inhibited the generation of p75highHNK1+/CD49D+ postmigratory neural crest stem cells (NCSCs) and upregulated the cell apoptosis rate during NC commitment from hiPSCs. Moreover, we discovered that both the neuronal and glial differentiation capacities of SOX10−/− NCSCs were severely compromised. Intriguingly, we showed that SOX10−/− hiPSCs generated markedly more TFAP2C+nonneural ectoderm cells (NNE) than control hiPSCs during neural crest differentiation. Our results indicate that SOX10 is crucial for the transition of premigratory cells to migrating NC and is vital for NC survival. Taken together, these results provide new insights into the function of SOX10 in human NC development, and the SOX10-knockout hiPSC lines may serve as a valuable cell model to study the pathogenesis of SOX10-related human neurocristopathies.
Highlights
Neural crest stem cells (NCSCs) are multipotent cells in the developing vertebrate embryo that give rise to a wide range of tissues and cell types, including the peripheral nervous system (PNS), enteric nervous system (ENS), craniofacial skeletal tissue, and melanocytes of the skin [1, 2]
In this study, we successfully established SRY-Box Transcription Factor 10 (SOX10)-knockout human induced pluripotent stem cell (hiPSC) and discovered that SOX10 deletion impaired neural crest survival and inhibited the generation of migratory neural crest from premigratory progenitors, as illustrated by the higher numbers of premigratory NCSCs and lower numbers of migrating NCSCs, decreased migration ability, upregulated apoptosis rate, and compromised multipotency in the SOX10-KO group compared to the WT group
We found that SOX10-knockout hESCs possessed similar phenotypes including postmigratory neural crest differentiation defects, and increased cell apoptosis rate and reactive oxygen species (ROS) production with SOX10-knockout hiPSCs during neural crest differentiation (Supplementary Fig. S10)
Summary
Neural crest stem cells (NCSCs) are multipotent cells in the developing vertebrate embryo that give rise to a wide range of tissues and cell types, including the peripheral nervous system (PNS), enteric nervous system (ENS), craniofacial skeletal tissue, and melanocytes of the skin [1, 2]. There have been numerous efforts to study neural crest development and their related diseases using human pluripotent stem cell (hPSC) models [8,9,10,11]. Several protocols have been established for the robust differentiation of NCSCs from hPSCs [12,13,14,15]. These methods provide us with valuable platforms for studying the cellular and molecular mechanisms involved in human NCSC development and neurocristopathies
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