title: Oral treatment with Cu(II)(atsm) increases mutant SOD1 in vivo but protects motor neurons and improves the phenotype of a transgenic mouse model of amyotrophic lateral sclerosis.
abstract: Mutations in the metallo-protein Cu/Zn-superoxide dismutase (SOD1) cause amyotrophic lateral sclerosis (ALS) in humans and an expression level-dependent phenotype in transgenic rodents. We show that oral treatment with the therapeutic agent diacetyl-bis(4-methylthiosemicarbazonato)copper(II) [Cu(II)(atsm)] increased the concentration of mutant SOD1 (SOD1G37R) in ALS model mice, but paradoxically improved locomotor function and survival of the mice. To determine why the mice with increased levels of mutant SOD1 had an improved phenotype, we analyzed tissues by mass spectrometry. These analyses revealed most SOD1 in the spinal cord tissue of the SOD1G37R mice was Cu deficient. Treating with Cu(II)(atsm) decreased the pool of Cu-deficient SOD1 and increased the pool of fully metallated (holo) SOD1. Tracking isotopically enriched (65)Cu(II)(atsm) confirmed the increase in holo-SOD1 involved transfer of Cu from Cu(II)(atsm) to SOD1, suggesting the improved locomotor function and survival of the Cu(II)(atsm)-treated SOD1G37R mice involved, at least in part, the ability of the compound to improve the Cu content of the mutant SOD1. This was supported by improved survival of SOD1G37R mice that expressed the human gene for the Cu uptake protein CTR1. Improving the metal content of mutant SOD1 in vivo with Cu(II)(atsm) did not decrease levels of misfolded SOD1. These outcomes indicate the metal content of SOD1 may be a greater determinant of the toxicity of the protein in mutant SOD1-associated forms of ALS than the mutations themselves. Improving the metal content of SOD1 therefore represents a valid therapeutic strategy for treating ALS caused by SOD1.
authors.affiliation: Florey Institute of Neuroscience and Mental Health.
authors.forename: Blaine R
authors.initials: BR
authors.lastname: Roberts
authors.affiliation: Department of Pathology.
authors.forename: Nastasia K H
authors.initials: NK
authors.lastname: Lim
authors.affiliation: Department of Pathology.
authors.forename: Erin J
authors.initials: EJ
authors.lastname: McAllum
authors.affiliation: School of Chemistry, Bio21 Institute of Biotechnology and Molecular Biology, and.
authors.forename: Paul S
authors.initials: PS
authors.lastname: Donnelly
authors.affiliation: Florey Institute of Neuroscience and Mental Health, Elemental Bio-imaging Facility, and Department of Chemistry and Forensic Science, University of Technology Sydney, 2007 Broadway, New South Wales, Australia.
authors.forename: Dominic J
authors.initials: DJ
authors.lastname: Hare
authors.affiliation: Elemental Bio-imaging Facility, and Department of Chemistry and Forensic Science, University of Technology Sydney, 2007 Broadway, New South Wales, Australia.
authors.forename: Philip A
authors.initials: PA
authors.lastname: Doble
authors.affiliation: Florey Institute of Neuroscience and Mental Health.
authors.forename: Bradley J
authors.initials: BJ
authors.lastname: Turner
authors.affiliation: Department of Pathology.
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Price
authors.affiliation: School of Chemistry, Bio21 Institute of Biotechnology and Molecular Biology, and.
authors.forename: Sin Chun
authors.initials: SC
authors.lastname: Lim
authors.affiliation: School of Chemistry, Bio21 Institute of Biotechnology and Molecular Biology, and.
authors.forename: Brett M
authors.initials: BM
authors.lastname: Paterson
authors.affiliation: School of Chemistry, Bio21 Institute of Biotechnology and Molecular Biology, and.
authors.forename: James L
authors.initials: JL
authors.lastname: Hickey
authors.affiliation: Department of Biochemistry and Biophysics, Linus Pauling Institute, and.
authors.forename: Timothy W
authors.initials: TW
authors.lastname: Rhoads
authors.affiliation: Department of Biochemistry and Biophysics, Linus Pauling Institute, and.
authors.forename: Jared R
authors.initials: JR
authors.lastname: Williams
authors.affiliation: Department of Pathology.
authors.forename: Katja M
authors.initials: KM
authors.lastname: Kanninen
authors.affiliation: Florey Institute of Neuroscience and Mental Health, Bio21 Institute of Biotechnology and Molecular Biology, and.
authors.forename: Lin W
authors.initials: LW
authors.lastname: Hung
authors.affiliation: Department of Pathology.
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Liddell
authors.affiliation: Department of Pathology.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Grubman
authors.affiliation: Centre for Cellular and Molecular Biology, School of Life and Environmental Sciences, Deakin University, Burwood, 3125 Victoria, Australia.
authors.forename: Jean-Francois
authors.initials: JF
authors.lastname: Monty
authors.affiliation: Centre for Cellular and Molecular Biology, School of Life and Environmental Sciences, Deakin University, Burwood, 3125 Victoria, Australia.
authors.forename: Roxana M
authors.initials: RM
authors.lastname: Llanos
authors.affiliation: ANU Medical School, ANU College of Medicine, Biology and the Environment, The Australian National University, 0200 Canberra, Australian Capital Territory, Australia, and.
authors.forename: David R
authors.initials: DR
authors.lastname: Kramer
authors.affiliation: Centre for Cellular and Molecular Biology, School of Life and Environmental Sciences, Deakin University, Burwood, 3125 Victoria, Australia.
authors.forename: Julian F B
authors.initials: JF
authors.lastname: Mercer
authors.affiliation: Florey Institute of Neuroscience and Mental Health.
authors.forename: Ashley I
authors.initials: AI
authors.lastname: Bush
authors.affiliation: Florey Institute of Neuroscience and Mental Health.
authors.forename: Colin L
authors.identifier: http://orcid.org/0000-0001-8259-9069
authors.initials: CL
authors.lastname: Masters
authors.affiliation: Florey Institute of Neuroscience and Mental Health, School of Molecular and Cellular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom.
authors.forename: James A
authors.initials: JA
authors.lastname: Duce
authors.affiliation: Florey Institute of Neuroscience and Mental Health.
authors.forename: Qiao-Xin
authors.initials: QX
authors.lastname: Li
authors.affiliation: Department of Biochemistry and Biophysics, Linus Pauling Institute, and Environmental Health Sciences Centre, Oregon State University, Corvallis, Oregon 97331.
authors.forename: Joseph S
authors.initials: JS
authors.lastname: Beckman
authors.affiliation: Florey Institute of Neuroscience and Mental Health, Bio21 Institute of Biotechnology and Molecular Biology, and Department of Pharmacology, The University of Melbourne, Parkville, 3010 Victoria, Australia.
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Barnham
authors.affiliation: Florey Institute of Neuroscience and Mental Health, Department of Pathology.
authors.forename: Anthony R
authors.initials: AR
authors.lastname: White
authors.affiliation: Florey Institute of Neuroscience and Mental Health, Department of Pathology, pjcrouch@unimelb.edu.au.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Crouch
mesh_terms: D000284:Administration, Oral; D000367:Age Factors; D000690:Amyotrophic Lateral Sclerosis / Q000188:drug therapy / Q000235:genetics / Q000401:mortality / Q000473:pathology; D000818:Animals; D027682:Cation Transport Proteins / Q000235:genetics; D002850:Chromatography, Gel; D056831:Coordination Complexes; D000079923:Copper Transporter 1; D004195:Disease Models, Animal; D006801:Humans; D008124:Locomotion / Q000187:drug effects / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D009046:Motor Neurons / Q000187:drug effects*; D009154:Mutation / Q000235:genetics*; D009942:Organometallic Compounds / Q000008:administration & dosage*; D010641:Phenotype; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism; D000072105:Superoxide Dismutase-1; D013882:Thiosemicarbazones / Q000008:administration & dosage*
chemical_list: D027682:Cation Transport Proteins; D056831:Coordination Complexes; D000079923:Copper Transporter 1; D009942:Organometallic Compounds; C490079:SLC31A1 protein, human; C000606290:SOD1 protein, human; D013882:Thiosemicarbazones; C089548:copper (II) diacetyl-di(N(4)-methylthiosemicarbazone); C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1