PC1/3 deficiency impacts POMC processing in human embryonic stem cell-derived hypothalamic neurons

We developed a technique for generating hypothalamic neurons from human pluripotent stem cells. Here, as proof-of-principle, we examine the use of these cells in modeling of a monogenic form of severe obesity: PCSK1 deficiency. We generated PCSK1 (PC1/3)-deficient human embryonic stem cell (hESC) lines using both shRNA and CRISPR-Cas9, and investigated pro-opiomelanocortin (POMC) processing using hESC-differentiated hypothalamic neurons. Overall design: We tried to idenitify transcripitional profiles and specific transcription factors that involved in of different stages during hypothalamic neuron differentiation from single cell sequencing for hESC-derived Day27 hypothalamic neurons, Day 12 neuron progenitors and undifferentiated stem cells

本研究建立了一套从人多能干细胞（human pluripotent stem cells）诱导分化下丘脑神经元的技术。本研究以此作为原理验证，探究了此类细胞在单基因遗传性重度肥胖模型——PCSK1缺乏症——中的应用。本研究通过短发夹RNA（shRNA）与CRISPR-Cas9两种方法，构建了PCSK1（PC1/3）缺陷型人胚胎干细胞（human embryonic stem cell, hESC）系，并利用hESC诱导分化的下丘脑神经元，对促阿黑皮素原（pro-opiomelanocortin, POMC）的加工过程进行了研究。实验设计：本研究针对hESC来源的第27天下丘脑神经元、第12天神经元祖细胞及未分化干细胞开展单细胞测序，旨在解析下丘脑神经元分化各阶段的转录组特征与特异性转录因子。