title: Arrested Hematopoiesis and Vascular Relaxation Defects in Mice with a Mutation in Dhfr.
abstract: Dihydrofolate reductase (DHFR) is a critical enzyme in the folate metabolism pathway and also plays a role in regulating nitric oxide (NO) signaling in endothelial cells. Although both coding and noncoding mutations with phenotypic effects have been identified in the human DHFR gene, no mouse model is currently available to study the consequences of perturbing DHFR in vivo In order to identify genes involved in definitive hematopoiesis, we performed a forward genetic screen and produced a mouse line, here referred to as Orana, with a point mutation in the Dhfr locus leading to a Thr136Ala substitution in the DHFR protein. Homozygote Orana mice initiate definitive hematopoiesis, but expansion of progenitors in the fetal liver is compromised, and the animals die between embryonic day 13.5 (E13.5) and E14.5. Heterozygote Orana mice survive to adulthood but have tissue-specific alterations in folate abundance and distribution, perturbed stress erythropoiesis, and impaired endothelium-dependent relaxation of the aorta consistent with the role of DHFR in regulating NO signaling. Orana mice provide insight into the dual roles of DHFR and are a useful model for investigating the role of environmental and dietary factors in the context of vascular defects caused by altered NO signaling.
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Julie A I
authors.initials: JA
authors.lastname: Thoms
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Kathy
authors.initials: K
authors.lastname: Knezevic
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Jia Jenny
authors.initials: JJ
authors.lastname: Liu
authors.affiliation: School of Medical Sciences, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Elias N
authors.initials: EN
authors.lastname: Glaros
authors.affiliation: School of Medical Sciences, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Thuan
authors.initials: T
authors.lastname: Thai
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Qiao
authors.initials: Q
authors.lastname: Qiao
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Heather
authors.initials: H
authors.lastname: Campbell
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Deborah
authors.initials: D
authors.lastname: Packham
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Yizhou
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Australian Phenomics Facility, John Curtin School of Medical Research, Australian National University, Acton, ACT, Australia.
authors.forename: Peter
authors.initials: P
authors.lastname: Papathanasiou
authors.affiliation: Australian Phenomics Facility, John Curtin School of Medical Research, Australian National University, Acton, ACT, Australia.
authors.forename: Robert
authors.initials: R
authors.lastname: Tunningley
authors.affiliation: Australian Phenomics Facility, John Curtin School of Medical Research, Australian National University, Acton, ACT, Australia.
authors.forename: Belinda
authors.initials: B
authors.lastname: Whittle
authors.affiliation: School of Medical Sciences, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Amanda W S
authors.initials: AW
authors.lastname: Yeung
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Vashe
authors.initials: V
authors.lastname: Chandrakanthan
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Luke
authors.initials: L
authors.lastname: Hesson
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia Haematology, Concord Repatriation and General Hospital, Concord, NSW, Australia.
authors.forename: Vivien
authors.initials: V
authors.lastname: Chen
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Jason W H
authors.initials: JW
authors.lastname: Wong
authors.affiliation: Stem Cell Regulation Unit, St. Vincent's Institute of Medical Research and Department of Medicine at St. Vincent's Hospital, The University of Melbourne, Fitzroy, Victoria, Australia.
authors.forename: Louise E
authors.initials: LE
authors.lastname: Purton
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Robyn L
authors.initials: RL
authors.lastname: Ward
authors.affiliation: School of Medical Sciences, University of New South Wales, Sydney, NSW, Australia shane.thomas@unsw.edu.au pimanda@unsw.edu.au.
authors.forename: Shane R
authors.initials: SR
authors.lastname: Thomas
authors.affiliation: Adult Cancer Program, Prince of Wales Clinical School, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia Department of Haematology, Prince of Wales Hospital, Sydney, NSW, Australia shane.thomas@unsw.edu.au pimanda@unsw.edu.au.
authors.forename: John E
authors.initials: JE
authors.lastname: Pimanda
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001011:Aorta / Q000502:physiology*; D001483:Base Sequence; D002460:Cell Line; D005492:Folic Acid / Q000378:metabolism; D006410:Hematopoiesis; D006720:Homozygote; D006801:Humans; D008099:Liver / Q000196:embryology / Q000378:metabolism; D051379:Mice / Q000196:embryology* / Q000235:genetics* / Q000502:physiology; D008810:Mice, Inbred C57BL; D008958:Models, Molecular; D009569:Nitric Oxide / Q000378:metabolism; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D009569:Nitric Oxide; D005492:Folic Acid; D013762:Tetrahydrofolate Dehydrogenase
references.citation: Blood. 2013 Feb 21;121(8):e43-9
references.pmid: 23287863
references.citation: FASEB J. 1990 Mar;4(5):1450-9
references.pmid: 2407589
references.citation: Diabetologia. 2012 Jul;55(7):2069-79
references.pmid: 22549734
references.citation: Circulation. 2005 Apr 26;111(16):2126-33
references.pmid: 15824200
references.citation: Cell. 2009 May 15;137(4):736-48
references.pmid: 19450519
references.citation: Hypertension. 2010 Dec;56(6):1137-44
references.pmid: 21059996
references.citation: Biomed Chromatogr. 2011 Apr;25(4):427-31
references.pmid: 21374646
references.citation: Nature. 1993 Jul 1;364(6432):64-7
references.pmid: 8316298
references.citation: Blood. 1995 Jan 15;85(2):500-9
references.pmid: 7812005
references.citation: Am J Hum Genet. 2011 Feb 11;88(2):216-25
references.pmid: 21310276
references.citation: Diabetes. 2007 Jan;56(1):118-26
references.pmid: 17192473
references.citation: Cell. 1996 Sep 20;86(6):897-906
references.pmid: 8808625
references.citation: Am J Med Genet A. 2004 Feb 1;124A(4):339-45
references.pmid: 14735580
references.citation: Anal Biochem. 1999 Aug 1;272(2):143-8
references.pmid: 10415082
references.citation: Nature. 2009 Jun 25;459(7250):1131-5
references.pmid: 19440194
references.citation: Biochemistry. 1990 Oct 9;29(40):9467-79
references.pmid: 2248959
references.citation: J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Jul 1;870(1):63-7
references.pmid: 18562255
references.citation: PLoS One. 2014;9(2):e88899
references.pmid: 24558445
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13481-6
references.pmid: 12359872
references.citation: Hypertension. 2012 Jan;59(1):158-66
references.pmid: 22083158
references.citation: Am J Hum Genet. 2011 Feb 11;88(2):226-31
references.pmid: 21310277
references.citation: Blood. 2010 Dec 23;116(26):5849-58
references.pmid: 20610815
references.citation: Nat Struct Mol Biol. 2013 Nov;20(11):1243-9
references.pmid: 24077226
references.citation: Blood. 1996 Mar 1;87(5):2065-70
references.pmid: 8634458
references.citation: Biochem J. 2000 Apr 1;347 Pt 1:1-16
references.pmid: 10727395
references.citation: Am J Med Genet A. 2007 Jun 1;143A(11):1174-80
references.pmid: 17486595
references.citation: Adv Genet. 2010;71:79-121
references.pmid: 20933127
references.citation: J Mol Cell Cardiol. 2009 Dec;47(6):752-60
references.pmid: 19660467
references.citation: Cardiovasc Res. 2012 Mar 15;93(4):694-701
references.pmid: 22241166
references.citation: Nat Genet. 2004 Dec;36(12):1296-300
references.pmid: 15516932
references.citation: Nucleic Acids Res. 2008 Oct;36(18):e119
references.pmid: 18718928
references.citation: Anal Bioanal Chem. 2008 May;391(1):263-70
references.pmid: 18368393
references.citation: Am J Hematol. 2009 Aug;84(8):526-9
references.pmid: 19536847
references.citation: Cancer Prev Res (Phila). 2012 Jul;5(7):921-9
references.pmid: 22609762
references.citation: Cell Stem Cell. 2009 Mar 6;4(3):263-74
references.pmid: 19265665
references.citation: Arterioscler Thromb Vasc Biol. 2013 Mar;33(3):459-65
references.pmid: 23307875
references.citation: Annu Rev Nutr. 2004;24:105-31
references.pmid: 15189115
references.citation: J Biol Chem. 2009 Oct 9;284(41):28128-36
references.pmid: 19666465
references.citation: Cell Stem Cell. 2008 Dec 4;3(6):625-36
references.pmid: 19041779
references.citation: J Biol Chem. 2005 Jun 17;280(24):22721-31
references.pmid: 15817466
references.citation: Proc Natl Acad Sci U S A. 2005 Jun 21;102(25):9056-61
references.pmid: 15941833
references.citation: Anal Biochem. 2008 Feb 15;373(2):383-5
references.pmid: 18028864
references.citation: Blood. 1995 Jul 15;86(2):677-84
references.pmid: 7605998
references.citation: Genetics. 1988 Feb;118(2):299-305
references.pmid: 3360305
references.citation: Proc Natl Acad Sci U S A. 1998 Aug 4;95(16):9220-5
references.pmid: 9689061
references.citation: Eur J Biochem. 1988 Jun 1;174(2):377-85
references.pmid: 3383852
references.citation: Circulation. 2005 Apr 26;111(16):2086-90
references.pmid: 15824199
references.citation: Am J Physiol Gastrointest Liver Physiol. 2011 Jan;300(1):G1-11
references.pmid: 20947703
references.citation: Antioxid Redox Signal. 2008 Oct;10(10):1713-65
references.pmid: 18707220
references.citation: Mol Cancer Ther. 2009 Feb;8(2):424-32
references.pmid: 19190117
references.citation: J Biol Chem. 2009 May 8;284(19):12691-700
references.pmid: 19286667
references.citation: Anal Biochem. 2011 Apr 15;411(2):210-7
references.pmid: 21192913
references.citation: Arterioscler Thromb Vasc Biol. 2001 Apr;21(4):496-502
references.pmid: 11304463
references.citation: Curr Genomics. 2010 Dec;11(8):578-83
references.pmid: 21629435
references.citation: Curr Drug Metab. 2002 Apr;3(2):159-73
references.pmid: 12003348