title: Fragments of HdhQ150 mutant huntingtin form a soluble oligomer pool that declines with aggregate deposition upon aging.
abstract: Cleavage of the full-length mutant huntingtin (mhtt) protein into smaller, soluble aggregation-prone mhtt fragments appears to be a key process in the neuropathophysiology of Huntington's Disease (HD). Recent quantification studies using TR-FRET-based immunoassays showed decreasing levels of soluble mhtt correlating with an increased load of aggregated mhtt in the aging HdhQ150 mouse brain. To better characterize the nature of these changes at the level of native mhtt species, we developed a detection method that combines size exclusion chromatography (SEC) and time-resolved fluorescence resonance energy transfer (TR-FRET) that allowed us to resolve and define the formation, aggregation and temporal dynamics of native soluble mhtt species and insoluble aggregates in the brain of the HdhQ150 knock-in mouse. We found that mhtt fragments and not full-length mhtt form oligomers in the brains of one month-old mice long before disease phenotypes and mhtt aggregate histopathology occur. As the HdhQ150 mice age, brain levels of soluble full-length mhtt protein remain similar. In contrast, the soluble oligomeric pool of mhtt fragments slightly increases during the first two months before it declines between 3 and 8 months of age. This decline inversely correlates with the formation of insoluble mhtt aggregates. We also found that the pool-size of soluble mhtt oligomers is similar in age-matched heterozygous and homozygous HdhQ150 mouse brains whereas insoluble aggregate formation is greatly accelerated in the homozygous mutant brain. The capacity of the soluble mhtt oligomer pool therefore seems exhausted already in the heterozygous state and likely kept constant by changes in flux and, as a consequence, increased rate of insoluble aggregate formation. We demonstrate that our novel findings in mice translate to human HD brain but not HD patient fibroblasts.
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Pharma AG, Basel, Switzerland.
authors.forename: David
authors.initials: D
authors.lastname: Marcellin
authors.forename: Dorothee
authors.initials: D
authors.lastname: Abramowski
authors.forename: Douglas
authors.initials: D
authors.lastname: Young
authors.forename: Jens
authors.initials: J
authors.lastname: Richter
authors.forename: Andreas
authors.initials: A
authors.lastname: Weiss
authors.forename: Audrey
authors.initials: A
authors.lastname: Marcel
authors.forename: Julia
authors.initials: J
authors.lastname: Maassen
authors.forename: Muriel
authors.initials: M
authors.lastname: Kauffmann
authors.forename: Miriam
authors.initials: M
authors.lastname: Bibel
authors.forename: Derya R
authors.initials: DR
authors.lastname: Shimshek
authors.forename: Richard L M
authors.initials: RL
authors.lastname: Faull
authors.forename: Gillian P
authors.initials: GP
authors.lastname: Bates
authors.forename: Rainer R
authors.initials: RR
authors.lastname: Kuhn
authors.forename: P Herman
authors.initials: PH
authors.lastname: Van der Putten
authors.forename: Peter
authors.initials: P
authors.lastname: Schmid
authors.forename: Gregor P
authors.initials: GP
authors.lastname: Lotz
mesh_terms: D000375:Aging; D000818:Animals; D001921:Brain / Q000378:metabolism / Q000473:pathology; D002845:Chromatography / Q000379:methods; D004195:Disease Models, Animal; D053595:Embryonic Stem Cells / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D006801:Humans; D000071058:Huntingtin Protein; D006816:Huntington Disease / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics*; D011485:Protein Binding; D050486:Serotonin Plasma Membrane Transport Proteins / Q000235:genetics* / Q000502:physiology
chemical_list: C086055:HTT protein, human; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins; D050486:Serotonin Plasma Membrane Transport Proteins; C493231:Slc6a4 protein, mouse