title: PC1/3 Deficiency Impacts Pro-opiomelanocortin Processing in Human Embryonic Stem Cell-Derived Hypothalamic Neurons.
abstract: 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.
authors.affiliation: 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.
authors.forename: Liheng
authors.initials: L
authors.lastname: Wang
authors.affiliation: 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.
authors.forename: Lina
authors.initials: L
authors.lastname: Sui
authors.affiliation: Department of Medicine and Naomi Berrie Diabetes Center, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
authors.forename: Sunil K
authors.initials: SK
authors.lastname: Panigrahi
authors.affiliation: Department of Medicine and Naomi Berrie Diabetes Center, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
authors.forename: Kana
authors.initials: K
authors.lastname: Meece
authors.affiliation: Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA.
authors.forename: Yurong
authors.initials: Y
authors.lastname: Xin
authors.affiliation: Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA.
authors.forename: Jinrang
authors.initials: J
authors.lastname: Kim
authors.affiliation: Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA.
authors.forename: Jesper
authors.initials: J
authors.lastname: Gromada
authors.affiliation: Department of Pathology and Cell Biology and Naomi Berrie Diabetes Center, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
authors.forename: Claudia A
authors.initials: CA
authors.lastname: Doege
authors.affiliation: Department of Medicine and Naomi Berrie Diabetes Center, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
authors.forename: Sharon L
authors.initials: SL
authors.lastname: Wardlaw
authors.affiliation: 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; New York Stem Cell Foundation Research Institute, 3960 Broadway, New York, NY 10032, USA.
authors.forename: Dieter
authors.initials: D
authors.lastname: Egli
authors.affiliation: 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. Electronic address: rl232@columbia.edu.
authors.forename: Rudolph L
authors.initials: RL
authors.lastname: Leibel
mesh_terms: D000324:Adrenocorticotropic Hormone / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D064113:CRISPR-Cas Systems; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D059865:Endoplasmic Reticulum Stress; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D018390:Gene Targeting; D000066449:Human Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D009154:Mutation; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D011333:Pro-Opiomelanocortin / Q000378:metabolism*; D045664:Proprotein Convertase 1 / Q000172:deficiency* / Q000235:genetics; D059748:Proteolysis; D017966:Pyramidal Cells / Q000166:cytology / Q000378:metabolism; D000521:alpha-MSH / Q000378:metabolism
chemical_list: D000521:alpha-MSH; D011333:Pro-Opiomelanocortin; D000324:Adrenocorticotropic Hormone; D045664:Proprotein Convertase 1
keywords: PC1/3 deficiency; POMC processing; hESC; hypothalamic neurons; obesity