Amin Neal D, Kelley Kevin W, Hao Jin, Miura Yuki, Narazaki Genta, Li Tommy, McQueen Patrick, Kulkarni Shravanti, Pavlov Sergey, Paşca Sergiu P
bioRxiv. 2023 Jun 1:2023.05.31.541819. doi: 10.1101/2023.05.31.541819.
Morphogens choreograph the generation of remarkable cellular diversity in the developing nervous system. Differentiation of stem cells toward particular neural cell fates often relies upon combinatorial modulation of these signaling pathways. However, the lack of a systematic approach to understand morphogen-directed differentiation has precluded the generation of many neural cell populations, and knowledge of the general principles of regional specification remain in-complete. Here, we developed an arrayed screen of 14 morphogen modulators in human neural organoids cultured for over 70 days. Leveraging advances in multiplexed RNA sequencing technology and annotated single cell references of the human fetal brain we discovered that this screening approach generated considerable regional and cell type diversity across the neural axis. By deconvoluting morphogen-cell type relationships, we extracted design principles of brain region specification, including critical morphogen timing windows and combinatorics yielding an array of neurons with distinct neuro-transmitter identities. Tuning GABAergic neural subtype diversity unexpectedly led to the derivation of primate-specific interneurons. Taken together, this serves as a platform towards an morphogen atlas of human neural cell differentiation that will bring insights into human development, evolution, and disease.
形态发生素在发育中的神经系统中编排了显著的细胞多样性的产生。干细胞向特定神经细胞命运的分化通常依赖于这些信号通路的组合调节。然而，缺乏一种系统的方法来理解形态发生素导向的分化阻碍了许多神经细胞群体的产生，并且区域特化的一般原则的知识仍然不完整。在这里，我们在培养超过70天的人类神经类器官中开发了一个包含14种形态发生素调节剂的阵列筛选。利用多重RNA测序技术的进展以及人类胎儿大脑的注释单细胞参考，我们发现这种筛选方法在整个神经轴上产生了相当大的区域和细胞类型多样性。通过反卷积形态发生素 - 细胞类型关系，我们提取了脑区特化的设计原则，包括关键的形态发生素时间窗口和组合方式，从而产生了一系列具有不同神经递质身份的神经元。调节GABA能神经亚型多样性意外地导致了灵长类特异性中间神经元的衍生。综上所述，这为构建人类神经细胞分化的形态发生素图谱提供了一个平台，该图谱将为人类发育、进化和疾病带来深刻见解。
