Abstract
Neural crest cells arise in the embryo from the neural plate border and migrate throughout the body, giving rise to many different tissue types such as bones and cartilage of the face, smooth muscles, neurons, and melanocytes. While studied extensively in animal models, neural crest development and disease have been poorly described in humans due to the challenges in accessing embryonic tissues. In recent years, patient-derived human induced pluripotent stem cells (hiPSCs) have become easier to generate, and several streamlined protocols have enabled robust differentiation of hiPSCs to the neural crest lineage. Thus, a unique opportunity is offered for modeling neurocristopathies using patient specific stem cell lines. In this work, we make use of hiPSCs derived from patients affected by the Bardet–Biedl Syndrome (BBS) ciliopathy. BBS patients often exhibit subclinical craniofacial dysmorphisms that are likely to be associated with the neural crest-derived facial skeleton. We focus on hiPSCs carrying variants in the BBS10 gene, which encodes a protein forming part of a chaperonin-like complex associated with the cilium. Here, we establish a pipeline for profiling hiPSCs during differentiation toward the neural crest stem cell fate. This can be used to characterize the differentiation properties of the neural crest-like cells. Two different BBS10 mutant lines showed a reduction in expression of the characteristic neural crest gene expression profile. Further analysis of both BBS10 mutant lines highlighted the inability of these mutant lines to differentiate toward a neural crest fate, which was also characterized by a decreased WNT and BMP response. Altogether, our study suggests a requirement for wild-type BBS10 in human neural crest development. In the long term, approaches such as the one we describe will allow direct comparison of disease-specific cell lines. This will provide valuable insights into the relationships between genetic background and heterogeneity in cellular models. The possibility of integrating laboratory data with clinical phenotypes will move us toward precision medicine approaches.
Highlights
Neural crest cells are a highly migratory, multipotent stem cell population that contributes to a broad range of tissues, including craniofacial bone and cartilage, peripheral neurons, glia, pigment, and other cells during embryonic development (Trainor, 2014)
As a proof of concept, we use the recently described (Leung et al, 2016) protocol to obtain efficient neural crest induction and compare HipSci cell lines from healthy volunteers with two cell lines obtained from Bardet–Biedl Syndrome (BBS) patients carrying BBS10 mutations (Table 1). We show that both mutant cell lines have altered neural crest induction and differentiation, demonstrating the feasibility of using human induced pluripotent stem cells (hiPSCs) from ciliopathies to explore processes affecting neural crest induction
The XIRY line carries two alleles: a non-synonymous change (V330A) on one allele, and a duplication on the other allele that leads to a frameshift and an early stop (T79Nfs∗17), likely to result in a loss-of-function variant (Figure 1B)
Summary
Neural crest cells are a highly migratory, multipotent stem cell population that contributes to a broad range of tissues, including craniofacial bone and cartilage, peripheral neurons, glia, pigment, and other cells during embryonic development (Trainor, 2014). Elucidating the cellular and molecular mechanisms of lineage specification and migration of neural crest stem cells is essential for understanding the pathogenesis of neurocristopathies, a family of diseases caused by anomalies in the migration or cell behavior of neural crest cells. Ciliopathies are a class of genetic disorders characterized by mutation of proteins affecting the structure or function of the cilium (Eggenschwiler and Anderson, 2007; Ishikawa and Marshall, 2011). Cranial ciliopathies are caused by mutations in ciliary genes that may result from altered neural crest cell development (Cortés et al, 2015). Bardet–Biedl Syndrome (BBS) is a prototypic ciliopathy that can affect neural crest-derived tissues
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