Generation of hypothalamic arcuate organoids from human induced pluripotent stem cells

Human brain organoids represent remarkable platforms for recapitulating features of human brain development and diseases. Existing organoid models do not resolve fine brain subregions, such as different nuclei in the hypothalamus. We report the generation of arcuate organoids (ARCOs) from human induced pluripotent stem cells (iPSCs) to model the development of the human hypothalamic arcuate nucleus. Single-cell RNA sequencing of ARCOs revealed significant molecular heterogeneity underlying different arcuate cell types, and machine learning-aided analysis based on the neonatal human hypothalamus single-nucleus transcriptome further showed a human arcuate nucleus molecular signature. We also explored ARCOs generated from Prader-Willi syndrome (PWS) patient iPSCs. These organoids exhibit aberrant differentiation and transcriptomic dysregulation similar to postnatal hypothalamus of PWS patients, indicative of cellular differentiation deficits and exacerbated inflammatory responses. Thus, patient iPSC-derived ARCOs represent a promising experimental model for investigating nucleus-specific features and disease-relevant mechanisms during early human arcuate development.

Automated summary

Human brain organoids derived from induced pluripotent stem cells (iPSCs) provide a valuable model for studying features of human brain development and diseases, such as the arcuate nucleus of the hypothalamus. By generating arcuate organoids (ARCOs) and analyzing their molecular heterogeneity, a human arcuate nucleus molecular signature was revealed. Furthermore, ARCOs derived from Prader-Willi syndrome (PWS) patient iPSCs showed aberrant differentiation and transcriptomic dysregulation, providing insights into disease mechanisms during early human arcuate development.