Wang Liheng, Sui Lina, Panigrahi Sunil K, Meece Kana, Xin Yurong, Kim Jinrang, Gromada Jesper, Doege Claudia A, Wardlaw Sharon L, Egli Dieter, Leibel Rudolph L
Division of Molecular Genetics, Department of Pediatrics and Naomi Berrie Diabetes Center, College of Physicians and Surgeons, Columbia University, 1150 St. Nicholas Avenue, Room 620A, New York, NY 10032, USA; Department of Medicine and Naomi Berrie Diabetes Center, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
Division of Molecular Genetics, Department of Pediatrics and Naomi Berrie Diabetes Center, College of Physicians and Surgeons, Columbia University, 1150 St. Nicholas Avenue, Room 620A, New York, NY 10032, USA.
Stem Cell Reports. 2017 Feb 14;8(2):264-277. doi: 10.1016/j.stemcr.2016.12.021. Epub 2017 Jan 26.
We recently 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. The cognate enzyme, PC1/3, processes many prohormones in neuroendocrine and other tissues. We generated PCSK1 (PC1/3)-deficient human embryonic stem cell (hESC) lines using both short hairpin RNA and CRISPR-Cas9, and investigated pro-opiomelanocortin (POMC) processing using hESC-differentiated hypothalamic neurons. The increased levels of unprocessed POMC and the decreased ratios (relative to POMC) of processed POMC-derived peptides in both PCSK1 knockdown and knockout hESC-derived neurons phenocopied POMC processing reported in PC1/3-null mice and PC1/3-deficient patients. PC1/3 deficiency was associated with increased expression of melanocortin receptors and PRCP (prolylcarboxypeptidase, a catabolic enzyme for α-melanocyte stimulating hormone (αMSH)), and reduced adrenocorticotropic hormone secretion. We conclude that the obesity accompanying PCSK1 deficiency may not be primarily due to αMSH deficiency.
我们最近开发了一种从人类多能干细胞生成下丘脑神经元的技术。在此，作为原理验证，我们研究了这些细胞在一种单基因形式的严重肥胖症（前蛋白转化酶枯草溶菌素/kexin 1型（PCSK1）缺乏症）模型中的应用。相关酶PC1/3在神经内分泌和其他组织中加工多种激素原。我们使用短发夹RNA和CRISPR-Cas9生成了PCSK1（PC1/3）缺乏的人类胚胎干细胞（hESC）系，并使用hESC分化的下丘脑神经元研究了阿黑皮素原（POMC）的加工过程。在PCSK1基因敲低和基因敲除的hESC衍生神经元中，未加工的POMC水平升高，加工后的POMC衍生肽与POMC的比率降低，这与PC1/3基因敲除小鼠和PC1/3缺乏患者中报道的POMC加工情况相似。PC1/3缺乏与黑皮质素受体和脯氨酰羧肽酶（PRCP，一种α-黑素细胞刺激激素（αMSH）的分解代谢酶）的表达增加以及促肾上腺皮质激素分泌减少有关。我们得出结论，PCSK1缺乏伴随的肥胖可能主要不是由于αMSH缺乏所致。
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