title: Single-step detection of mutant huntingtin in animal and human tissues: a bioassay for Huntington's disease.
abstract: The genetic mutation causing Huntington's disease is a polyglutamine expansion in the huntingtin protein where more than 37 glutamines cause disease by formation of toxic intracellular fragments, aggregates, and cell death. Despite a clear pathogenic role for mutant huntingtin, understanding huntingtin expression during the presymptomatic phase of the disease or during disease progression has remained obscure. Central to clarifying the role in the pathomechanism of disease is the ability to easily and accurately measure mutant huntingtin in accessible human tissue samples as well as cell and animal models. Here we describe a highly sensitive time-resolved Förster resonance energy transfer (FRET) assay for quantification of soluble mutant huntingtin in brain, plasma, and cerebrospinal fluid. Surprisingly, in mice, soluble huntingtin levels decrease during disease progression, inversely correlating with brain aggregate load. Mutant huntingtin is easily detected in human brain and blood-derived fractions, providing a utility to assess mutant huntingtin expression during disease course as well as a pharmacodynamic marker for disease-modifying therapeutics targeting expression, cleavage, or degradation of mutant huntingtin. The design of the homogeneous one-step method for huntingtin detection is such that it can be easily applied to measure other proteins of interest.
authors: Novartis Institutes for BioMedical Research, Basel, Switzerland.
authors: Andreas
authors: A
authors: Weiss
authors: Dorothée
authors: D
authors: Abramowski
authors: Miriam
authors: M
authors: Bibel
authors: Ruth
authors: R
authors: Bodner
authors: Vanita
authors: V
authors: Chopra
authors: Marian
authors: M
authors: DiFiglia
authors: Jonathan
authors: J
authors: Fox
authors: Kimberly
authors: K
authors: Kegel
authors: Corinna
authors: C
authors: Klein
authors: Stephan
authors: S
authors: Grueninger
authors: Steven
authors: S
authors: Hersch
authors: David
authors: D
authors: Housman
authors: Etienne
authors: E
authors: Régulier
authors: H Diana
authors: HD
authors: Rosas
authors: Muriel
authors: M
authors: Stefani
authors: Scott
authors: S
authors: Zeitlin
authors: Graeme
authors: G
authors: Bilbe
authors: Paolo
authors: P
authors: Paganetti
mesh_terms: D000328:Adult; D000704:Analysis of Variance; D000818:Animals; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D018450:Disease Progression; D053595:Embryonic Stem Cells / Q000378:metabolism; D005091:Exons; D005260:Female; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D015870:Gene Expression; D006801:Humans; D000071058:Huntingtin Protein; D006816:Huntington Disease / Q000097:blood / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008875:Middle Aged; D018482:Muscle, Skeletal / Q000378:metabolism; D050505:Mutant Proteins / Q000302:isolation & purification* / Q000378:metabolism; D019579:Neocortex / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D012680:Sensitivity and Specificity; D013997:Time Factors; D055815:Young Adult
chemical_list: C086055:HTT protein, human; C506812:Htt protein, mouse; D000071058:Huntingtin Protein; D050505:Mutant Proteins; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins