Laboratory of Clinical Biotechnology, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo 113-8655, Japan; Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo 113-8655, Japan.
Laboratory of Clinical Biotechnology, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo 113-8655, Japan; Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo 113-8655, Japan.
Cell Rep. 2023 Apr 25;42(4):112276. doi: 10.1016/j.celrep.2023.112276. Epub 2023 Mar 24.
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.
虽然骨骼对于运动、内分泌功能和造血至关重要，但人类骨骼发育的分子机制仍有待阐明。在这里，我们介绍了一种综合方法，通过将人类多能干细胞体外诱导的体节与免疫缺陷小鼠肾囊下植入体节诱导体内骺软骨内骨形成相结合，来模拟人类骨骼发育。组织学和 scRNA-seq 分析表明，诱导的骨骼再现了软骨内骨化，由人类骨骼细胞和小鼠循环细胞组成。骨骼细胞类型及其轨迹与人类胚胎相似。单细胞多组学分析揭示了与构成细胞类型特异性基因调控网络（GRN）的多个转录因子相关的染色质可及性的动态变化。我们进一步鉴定了 ZEB2，它可能调节人类成骨中的 GRN。总的来说，这些结果确定了人类骨骼发育中 GRN 的组成部分，并为其研究提供了一个有价值的模型。
