title: Mildly compromised tetrahydrobiopterin cofactor biosynthesis due to Pts variants leads to unusual body fat distribution and abdominal obesity in mice.
abstract: Tetrahydrobiopterin (BH4) is an essential cofactor for the aromatic amino acid hydroxylases, alkylglycerol monooxygenase, and nitric oxide synthases (NOS). Inborn errors of BH4 metabolism lead to severe insufficiency of brain monoamine neurotransmitters while augmentation of BH4 by supplementation or stimulation of its biosynthesis is thought to ameliorate endothelial NOS (eNOS) dysfunction, to protect from (cardio-) vascular disease and/or prevent obesity and development of the metabolic syndrome. We have previously reported that homozygous knock-out mice for the 6-pyruvolytetrahydropterin synthase (PTPS; Pts-ko/ko) mice with no BH4 biosynthesis die after birth. Here we generated a Pts-knock-in (Pts-ki) allele expressing the murine PTPS-p.Arg15Cys with low residual activity (15% of wild-type in vitro) and investigated homozygous (Pts-ki/ki) and compound heterozygous (Pts-ki/ko) mutants. All mice showed normal viability and depending on the severity of the Pts alleles exhibited up to 90% reduction of PTPS activity concomitant with neopterin elevation and mild reduction of total biopterin while blood L-phenylalanine and brain monoamine neurotransmitters were unaffected. Yet, adult mutant mice with compromised PTPS activity (i.e., Pts-ki/ko, Pts-ki/ki or Pts-ko/wt) had increased body weight and elevated intra-abdominal fat. Comprehensive phenotyping of Pts-ki/ki mice revealed alterations in energy metabolism with proportionally higher fat content but lower lean mass, and increased blood glucose and cholesterol. Transcriptome analysis indicated changes in glucose and lipid metabolism. Furthermore, differentially expressed genes associated with obesity, weight loss, hepatic steatosis, and insulin sensitivity were consistent with the observed phenotypic alterations. We conclude that reduced PTPS activity concomitant with mildly compromised BH4-biosynthesis leads to abnormal body fat distribution and abdominal obesity at least in mice. This study associates a novel single gene mutation with monogenic forms of obesity.
authors.affiliation: Division of Metabolism, University Children's Hospital Zürich, Steinwiesstrasse 75, CH-8032, Zürich, Switzerland.|Affiliated with the Neuroscience Center Zurich (ZNZ), University of Zurich and ETH Zurich, Zürich, Switzerland.|Children's Research Center (CRC), Zürich, Switzerland.
authors.forename: Germaine
authors.initials: G
authors.lastname: Korner
authors.affiliation: Division of Metabolism, University Children's Hospital Zürich, Steinwiesstrasse 75, CH-8032, Zürich, Switzerland.|Affiliated with the Neuroscience Center Zurich (ZNZ), University of Zurich and ETH Zurich, Zürich, Switzerland.|Children's Research Center (CRC), Zürich, Switzerland.
authors.forename: Tanja
authors.initials: T
authors.lastname: Scherer
authors.affiliation: Division of Metabolism, University Children's Hospital Zürich, Steinwiesstrasse 75, CH-8032, Zürich, Switzerland.|Affiliated with the Neuroscience Center Zurich (ZNZ), University of Zurich and ETH Zurich, Zürich, Switzerland.|Children's Research Center (CRC), Zürich, Switzerland.
authors.forename: Dea
authors.initials: D
authors.lastname: Adamsen
authors.affiliation: Division of Metabolism, University Children's Hospital Zürich, Steinwiesstrasse 75, CH-8032, Zürich, Switzerland.
authors.forename: Alexander
authors.initials: A
authors.lastname: Rebuffat
authors.affiliation: BHF Centre of Excellence, Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, OX3 9DU, Oxford, UK.
authors.forename: Mark
authors.initials: M
authors.lastname: Crabtree
authors.affiliation: Division of Clinical Chemistry and Biochemistry, University Children's Hospital Zürich, Zürich, Switzerland.
authors.forename: Anahita
authors.initials: A
authors.lastname: Rassi
authors.affiliation: Division of Metabolism, University Children's Hospital Zürich, Steinwiesstrasse 75, CH-8032, Zürich, Switzerland.
authors.forename: Rossana
authors.initials: R
authors.lastname: Scavelli
authors.affiliation: Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Daigo
authors.initials: D
authors.lastname: Homma
authors.affiliation: Division of Animal Facility, University of Zurich, Zürich, Switzerland.
authors.forename: Birgit
authors.initials: B
authors.lastname: Ledermann
authors.affiliation: Division of Pediatric Endocrinology and Diabetology, University Children's Hospital Zürich, Zürich, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Konrad
authors.affiliation: Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ichinose
authors.affiliation: Institute of Food Nutrition and Health, Swiss Federal Institute of Technology Zürich, Zürich, Switzerland.
authors.forename: Christian
authors.initials: C
authors.lastname: Wolfrum
authors.affiliation: German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany.
authors.forename: Marion
authors.initials: M
authors.lastname: Horsch
authors.affiliation: German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany.|Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Str. 25, 81377, Munich, Germany.|German Center for Diabetes Research (DZD), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany.
authors.forename: Birgit
authors.initials: B
authors.lastname: Rathkolb
authors.affiliation: Molecular Nutritional Medicine, Else Kröner-Fresenius Center, Technische Universität München, Am Forum 8, 85354, Freising-Weihenstephan, Germany.|ZIEL - Center for Nutrition and Food Sciences, Technische Universität München, D-85350, Freising, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Klingenspor
authors.affiliation: German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany.|Chair of Experimental Genetics, Center of Life and Food Sciences Weihenstephan, Technische Universität München, D-85354, Freising-Weihenstephan, Germany.|German Center for Diabetes Research (DZD), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Beckers
authors.affiliation: Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Str. 25, 81377, Munich, Germany.
authors.forename: Eckhard
authors.initials: E
authors.lastname: Wolf
authors.affiliation: German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany.
authors.forename: Valérie
authors.initials: V
authors.lastname: Gailus-Durner
authors.affiliation: German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany.
authors.forename: Helmut
authors.initials: H
authors.lastname: Fuchs
authors.affiliation: German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany.|Chair of Experimental Genetics, Center of Life and Food Sciences Weihenstephan, Technische Universität München, D-85354, Freising-Weihenstephan, Germany.|German Center for Diabetes Research (DZD), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Hrabě de Angelis
authors.affiliation: Division of Metabolism, University Children's Hospital Zürich, Steinwiesstrasse 75, CH-8032, Zürich, Switzerland. nenad.blau@med.uni-heidelberg.de.|Dietmar-Hopp Metabolic Center, University Children's Hospital Heidelberg, Im Neuenheimer Feld 669, D-69120, Heidelberg, Germany. nenad.blau@med.uni-heidelberg.de.
authors.forename: Nenad
authors.initials: N
authors.lastname: Blau
authors.affiliation: German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany. jan.rozman@helmholtz-muenchen.de.|Molecular Nutritional Medicine, Else Kröner-Fresenius Center, Technische Universität München, Am Forum 8, 85354, Freising-Weihenstephan, Germany. jan.rozman@helmholtz-muenchen.de.|German Center for Diabetes Research (DZD), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany. jan.rozman@helmholtz-muenchen.de.
authors.forename: Jan
authors.initials: J
authors.lastname: Rozman
authors.affiliation: Division of Metabolism, University Children's Hospital Zürich, Steinwiesstrasse 75, CH-8032, Zürich, Switzerland. beat.thony@kispi.uzh.ch.|Affiliated with the Neuroscience Center Zurich (ZNZ), University of Zurich and ETH Zurich, Zürich, Switzerland. beat.thony@kispi.uzh.ch.|Children's Research Center (CRC), Zürich, Switzerland. beat.thony@kispi.uzh.ch.
authors.forename: Beat
authors.initials: B
authors.lastname: Thöny
mesh_terms: D000273:Adipose Tissue / Q000378:metabolism*; D000483:Alleles; D000818:Animals; D001708:Biopterins / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics; D050218:Body Fat Distribution; D001835:Body Weight / Q000235:genetics; D002784:Cholesterol / Q000235:genetics; D005260:Female; D005838:Genotype; D005947:Glucose / Q000235:genetics; D006579:Heterozygote; D006720:Homozygote; D050356:Lipid Metabolism / Q000235:genetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation / Q000235:genetics; D019001:Nitric Oxide Synthase / Q000235:genetics; D052250:Nitric Oxide Synthase Type III / Q000235:genetics; D056128:Obesity, Abdominal / Q000235:genetics*; D010649:Phenylalanine / Q000235:genetics; D019761:Phosphorus-Oxygen Lyases / Q000235:genetics*; D059467:Transcriptome / Q000235:genetics
chemical_list: D001708:Biopterins; D010649:Phenylalanine; D002784:Cholesterol; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; D019761:Phosphorus-Oxygen Lyases; C043497:6-pyruvoyltetrahydropterin synthase; C003402:sapropterin; D005947:Glucose
references.citation: Arterioscler Thromb Vasc Biol. 2013 Nov;33(11):2501-8
references.pmid: 24030551
references.citation: Toxicology. 2013 Jul 5;309:100-6
references.pmid: 23692951
references.citation: Methods Mol Biol. 2009;530:463-509
references.pmid: 19266331
references.citation: Biochem Biophys Res Commun. 2010 Sep 10;400(1):72-7
references.pmid: 20696134
references.citation: Curr Vasc Pharmacol. 2011 Mar;9(2):188-99
references.pmid: 21143176
references.citation: Arterioscler Thromb Vasc Biol. 2004 Mar;24(3):405-12
references.pmid: 14656742
references.citation: Biochem J. 2011 Sep 15;438(3):397-414
references.pmid: 21867484
references.citation: Circulation. 2001 Jul 17;104(3):342-5
references.pmid: 11457755
references.citation: Nat Methods. 2005 Jun;2(6):403-4
references.pmid: 15908916
references.citation: Diabetes. 2014 Oct;63(10):3557-64
references.pmid: 24848070
references.citation: Eur J Nucl Med Mol Imaging. 2005 Jan;32(1):84-91
references.pmid: 15290118
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: Am J Hum Genet. 1994 May;54(5):782-92
references.pmid: 8178819
references.citation: J Biol Chem. 1995 Dec 8;270(49):29498-506
references.pmid: 7493990
references.citation: Biochem J. 2000 Apr 1;347 Pt 1:1-16
references.pmid: 10727395
references.citation: Nature. 2006 Dec 14;444(7121):881-7
references.pmid: 17167477
references.citation: Int J Genomics. 2013;2013:609748
references.pmid: 23986905
references.citation: Biochem J. 2000 Jul 1;349(Pt 1):353-6
references.pmid: 10861247
references.citation: J Neurochem. 2003 Aug;86(3):615-26
references.pmid: 12859675
references.citation: Endocrinology. 2013 Feb;154(2):685-97
references.pmid: 23321697
references.citation: PLoS One. 2012;7(10):e46986
references.pmid: 23056550
references.citation: Cell Biochem Biophys. 2004;41(3):415-34
references.pmid: 15509890
references.citation: Lancet. 2010 Oct 23;376(9750):1417-27
references.pmid: 20971365
references.citation: Arterioscler Thromb Vasc Biol. 2011 Jun;31(6):1283-90
references.pmid: 21474828
references.citation: Circulation. 2006 Apr 4;113(13):1708-14
references.pmid: 16585403
references.citation: Nat Med. 2012 Jun;18(6):943-9
references.pmid: 22581287
references.citation: J Clin Pathol. 1976 May;29(5):444-51
references.pmid: 932231
references.citation: J Clin Invest. 2003 Sep;112(5):725-35
references.pmid: 12952921
references.citation: Diabetes. 2013 May;62(5):1478-89
references.pmid: 23349480
references.citation: Am J Physiol Endocrinol Metab. 2004 Nov;287(5):E919-25
references.pmid: 15265759
references.citation: J Biol Chem. 2001 Nov 2;276(44):41150-60
references.pmid: 11517215
references.citation: FASEB J. 2004 Dec;18(15):1900-2
references.pmid: 15467010
references.citation: Eur J Pharmacol. 2007 Jan 19;555(1):48-53
references.pmid: 17098227
references.citation: Annu Rev Nutr. 2002;22:61-86
references.pmid: 12055338
references.citation: Br J Pharmacol. 2003 Oct;140(4):701-6
references.pmid: 14534153
references.citation: FEBS Lett. 2004 Nov 19;577(3):507-11
references.pmid: 15556637
references.citation: Annu Rev Med. 2005;56:443-58
references.pmid: 15660521
references.citation: Circulation. 2007 Jul 3;116(1):39-48
references.pmid: 17576866
references.citation: Am J Clin Nutr. 1985 Jan;41(1):110-2
references.pmid: 3966413
references.citation: J Endocrinol. 2012 Jun;213(3):251-61
references.pmid: 22457517
references.citation: Cell Cycle. 2012 May 15;11(10):1918-28
references.pmid: 22544329
references.citation: Biol Chem. 1998 Dec;379(12):1441-7
references.pmid: 9894812
references.citation: Am J Med. 2007 Mar;120(3 Suppl 1):S12-8
references.pmid: 17320517
references.citation: Mol Cell Biochem. 2013 Jul;379(1-2):123-31
references.pmid: 23564066
references.citation: Thromb Haemost. 2012 Nov;108(5):832-9
references.pmid: 23052970
references.citation: EMBO J. 2013 Jan 9;32(1):45-59
references.pmid: 23178591
references.citation: Am J Physiol Heart Circ Physiol. 2003 Aug;285(2):H875-82
references.pmid: 12730050
references.citation: J Biol Chem. 2003 Jul 25;278(30):28303-11
references.pmid: 12734191
references.citation: Brain Res. 2007 Jan 5;1127(1):136-50
references.pmid: 17112485