Waisman Center, University of Wisconsin-Madison, Madison, WI, USA.
State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Chemistry and Biomedicine Innovation Center (ChemBIC), Nanjing University, Nanjing, China.
Nat Biotechnol. 2024 Sep;42(9):1404-1416. doi: 10.1038/s41587-023-01977-4. Epub 2023 Nov 16.
Central norepinephrine (NE) neurons, located mainly in the locus coeruleus (LC), are implicated in diverse psychiatric and neurodegenerative diseases and are an emerging target for drug discovery. To facilitate their study, we developed a method to generate 40-60% human LC-NE neurons from human pluripotent stem cells. The approach depends on our identification of ACTIVIN A in regulating LC-NE transcription factors in dorsal rhombomere 1 (r1) progenitors. In vitro generated human LC-NE neurons display extensive axonal arborization; release and uptake NE; and exhibit pacemaker activity, calcium oscillation and chemoreceptor activity in response to CO. Single-nucleus RNA sequencing (snRNA-seq) analysis at multiple timepoints confirmed NE cell identity and revealed the differentiation trajectory from hindbrain progenitors to NE neurons via an ASCL1-expressing precursor stage. LC-NE neurons engineered with an NE sensor reliably reported extracellular levels of NE. The availability of functional human LC-NE neurons enables investigation of their roles in psychiatric and neurodegenerative diseases and provides a tool for therapeutics development.
中枢去甲肾上腺素 (NE) 神经元主要位于蓝斑 (LC)，与多种精神疾病和神经退行性疾病有关，是药物发现的新兴靶点。为了便于研究，我们开发了一种方法，可从人类多能干细胞中产生 40-60%的人类 LC-NE 神经元。该方法依赖于我们鉴定出 ACTIVIN A 在调节背侧菱形 1 (r1) 祖细胞中的 LC-NE 转录因子。体外生成的人类 LC-NE 神经元表现出广泛的轴突分支；释放和摄取 NE；并表现出起搏活动、钙振荡和对 CO 的化学感受器活性。多个时间点的单细胞 RNA 测序 (snRNA-seq) 分析证实了 NE 细胞的身份，并揭示了通过表达 ASCL1 的前体细胞阶段从后脑祖细胞向 NE 神经元分化的轨迹。用 NE 传感器工程化的 LC-NE 神经元可靠地报告了细胞外 NE 的水平。功能性人类 LC-NE 神经元的可用性使人们能够研究它们在精神疾病和神经退行性疾病中的作用，并为治疗药物的开发提供了工具。
