Stem cell-based modeling and single-cell multiomics reveal gene-regulatory mechanisms underlying human skeletal development

• Published in: Cell reports (Cambridge) 2023-04, Vol.42 (4), p.112276-112276, Article 112276
• Main Authors: Tani, Shoichiro, Okada, Hiroyuki, Onodera, Shoko, Chijimatsu, Ryota, Seki, Masahide, Suzuki, Yutaka, Xin, Xiaonan, Rowe, David W., Saito, Taku, Tanaka, Sakae, Chung, Ung-il, Ohba, Shinsuke, Hojo, Hironori
• Format: Article
• Language: English
• Subjects: gene regulatory networks; human pluripotent stem cells; single-cell multiome analysis; skeletal development; ZEB2; zinc finger E-box binding homeobox 2
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2023.112276

Although the skeleton is essential for locomotion, endocrine functions, and hematopoiesis, the molecular mechanisms of human skeletal development remain to be elucidated. Here, we introduce an integrative method to model human skeletal development by combining in vitro sclerotome induction from human pluripotent stem cells and in vivo endochondral bone formation by implanting the sclerotome beneath the renal capsules of immunodeficient mice. Histological and scRNA-seq analyses reveal that the induced bones recapitulate endochondral ossification and are composed of human skeletal cells and mouse circulatory cells. The skeletal cell types and their trajectories are similar to those of human embryos. Single-cell multiome analysis reveals dynamic changes in chromatin accessibility associated with multiple transcription factors constituting cell-type-specific gene-regulatory networks (GRNs). We further identify ZEB2, which may regulate the GRNs in human osteogenesis. Collectively, these results identify components of GRNs in human skeletal development and provide a valuable model for its investigation. [Display omitted] •This paper reports a method to model bone development using human pluripotent stem cells•Induced bones recapitulate endochondral ossification processes at embryonic stages•Single-cell analyses reveal cell-type-specific gene-regulatory networks (GRNs)•The transcription factor ZEB2 regulates osteogenic GRNs Tani et al. develop a method to model endochondral bone formation at embryonic stages with human pluripotent stem cells. They investigate the dynamics of cell-type-specific gene-regulatory networks underlying human skeletal development by single-cell multiomics and identify a transcription factor, ZEB2, which may regulate the regulatory networks in osteogenesis.