Regulatory mechanisms underlying the development of neural crest cells and associated diseases as revealed by single-cell RNA sequencing

Abstract

Neural crest (NC) is a transient population of stem cells unique to vertebrates and that give rise to a wide varieties of cell types contributing to the development of various tissue organs. Hirschsprung disease (HSCR) is the most common neurocristopathy and is frequently associated with defects in the heart and/or the central nervous system (CNS), caused by the problems in the development of enteric (ENCCs) and cardiac (CNCCs) NC cells or perturbations of some pathway genes shared among these systems. Previous genetic screenings of human patients have identified various genes associated with these diseases, such as KIF7 (Kinesin Family Member 7), VCL (Vinculin) and PHOX2B (Paired-like homeobox 2b). However, it remains elusive how these genes involved in the development of nervous systems and heart, as well as the molecular basis underlying different disease states. In this study, I performed single-cell RNA sequencing (scRNA-seq) data analysis based on various disease models, including mouse mutants, human induced pluripotent stem cell (hPSC)-derived cell- and organoid- models, to dissect the disease state heterogeneity, lineage differentiation abnormality and their underlying molecular basis during the development of ENCCs, CNCCs and CNS neurons. Putative key regulators were prioritized and functionally validated, which could provide potential drug targets for the disease treatment in the future. The computational analysis pipeline for the scRNA-seq data derived from different cell lineages and developmental stages, enriched/mix cell populations, in vitro and in vivo system was optimised. Novel algorithms and strategies were developed to address specific biological questions, to yield novel insights for the regulatory mechanisms underlying various neurocristopathies.