BACKGROUND: Quantification of disease-associated proteins in the cerebrospinal fluid (CSF) has been critical for the study and treatment of several neurodegenerative disorders; however, mutant huntingtin protein (mHTT), the cause of Huntington's disease (HD), is at very low levels in CSF and, to our knowledge, has never been measured previously. METHODS: We developed an ultrasensitive single-molecule counting (SMC) mHTT immunoassay that was used to quantify mHTT levels in CSF samples from individuals bearing the HD mutation and from control individuals in 2 independent cohorts. RESULTS: This SMC mHTT immunoassay demonstrated high specificity for mHTT, high sensitivity with a femtomolar detection threshold, and a broad dynamic range. Analysis of the CSF samples showed that mHTT was undetectable in CSF from all controls but quantifiable in nearly all mutation carriers. The mHTT concentration in CSF was approximately 3-fold higher in patients with manifest HD than in premanifest mutation carriers. Moreover, mHTT levels increased as the disease progressed and were associated with 5-year onset probability. The mHTT concentration independently predicted cognitive and motor dysfunction. Furthermore, the level of mHTT was associated with the concentrations of tau and neurofilament light chain in the CSF, suggesting a neuronal origin for the detected mHTT. CONCLUSIONS: We have demonstrated that mHTT can be quantified in CSF from HD patients using the described SMC mHTT immunoassay. Moreover, the level of mHTT detected is associated with proximity to disease onset and diminished cognitive and motor function. The ability to quantify CSF mHTT will facilitate the study of HD, and mHTT quantification could potentially serve as a biomarker for the development and testing of experimental mHTT-lowering therapies for HD. TRIAL REGISTRATION: Not applicable. FUNDING: CHDI Foundation Inc.; Medical Research Council (MRC) UK; National Institutes for Health Research (NIHR); Rosetrees Trust; Swedish Research Council; and Knut and Alice Wallenberg Foundation.
BACKGROUND: Quantification of disease-associated proteins in the cerebrospinal fluid (CSF) has been critical for the study and treatment of several neurodegenerative disorders; however, mutant huntingtin protein (mHTT), the cause of Huntington's disease (HD), is at very low levels in CSF and, to our knowledge, has never been measured previously. METHODS: We developed an ultrasensitive single-molecule counting (SMC) mHTT immunoassay that was used to quantify mHTT levels in CSF samples from individuals bearing the HD mutation and from control individuals in 2 independent cohorts. RESULTS: This SMC mHTT immunoassay demonstrated high specificity for mHTT, high sensitivity with a femtomolar detection threshold, and a broad dynamic range. Analysis of the CSF samples showed that mHTT was undetectable in CSF from all controls but quantifiable in nearly all mutation carriers. The mHTT concentration in CSF was approximately 3-fold higher in patients with manifest HD than in premanifest mutation carriers. Moreover, mHTT levels increased as the disease progressed and were associated with 5-year onset probability. The mHTT concentration independently predicted cognitive and motor dysfunction. Furthermore, the level of mHTT was associated with the concentrations of tau and neurofilament light chain in the CSF, suggesting a neuronal origin for the detected mHTT. CONCLUSIONS: We have demonstrated that mHTT can be quantified in CSF from HDpatients using the described SMC mHTT immunoassay. Moreover, the level of mHTT detected is associated with proximity to disease onset and diminished cognitive and motor function. The ability to quantify CSF mHTT will facilitate the study of HD, and mHTT quantification could potentially serve as a biomarker for the development and testing of experimental mHTT-lowering therapies for HD. TRIAL REGISTRATION: Not applicable. FUNDING: CHDI Foundation Inc.; Medical Research Council (MRC) UK; National Institutes for Health Research (NIHR); Rosetrees Trust; Swedish Research Council; and Knut and Alice Wallenberg Foundation.
Authors: Piotr Lewczuk; Georg Beck; Hermann Esselmann; Ralf Bruckmoser; Rüdiger Zimmermann; Magdalena Fiszer; Mirko Bibl; Juan Manuel Maler; Johannes Kornhuber; Jens Wiltfang Journal: Clin Chem Date: 2006-02 Impact factor: 8.327
Authors: John Todd; Bob Freese; Ann Lu; Douglas Held; Jennifer Morey; Richard Livingston; Philippe Goix Journal: Clin Chem Date: 2007-09-21 Impact factor: 8.327
Authors: H Telenius; B Kremer; Y P Goldberg; J Theilmann; S E Andrew; J Zeisler; S Adam; C Greenberg; E J Ives; L A Clarke Journal: Nat Genet Date: 1994-04 Impact factor: 38.330
Authors: Nancy S Wexler; Judith Lorimer; Julie Porter; Fidela Gomez; Carol Moskowitz; Edith Shackell; Karen Marder; Graciela Penchaszadeh; Simone A Roberts; Javier Gayán; Denise Brocklebank; Stacey S Cherny; Lon R Cardon; Jacqueline Gray; Stephen R Dlouhy; Sandra Wiktorski; Marion E Hodes; P Michael Conneally; Jack B Penney; James Gusella; Jang-Ho Cha; Michael Irizarry; Diana Rosas; Steven Hersch; Zane Hollingsworth; Marcy MacDonald; Anne B Young; J Michael Andresen; David E Housman; Margot Mieja De Young; Ernesto Bonilla; Theresa Stillings; Americo Negrette; S Robert Snodgrass; Maria Dolores Martinez-Jaurrieta; Maria A Ramos-Arroyo; Jacqueline Bickham; Juan Sanchez Ramos; Frederick Marshall; Ira Shoulson; Gustavo J Rey; Andrew Feigin; Norman Arnheim; Amarilis Acevedo-Cruz; Leticia Acosta; Jose Alvir; Kenneth Fischbeck; Leslie M Thompson; Angela Young; Leon Dure; Christopher J O'Brien; Jane Paulsen; Adam Brickman; Denise Krch; Shelley Peery; Penelope Hogarth; Donald S Higgins; Bernhard Landwehrmeyer Journal: Proc Natl Acad Sci U S A Date: 2004-03-01 Impact factor: 11.205
Authors: Annette Dalrymple; Edward J Wild; Richard Joubert; Kirupa Sathasivam; Maria Björkqvist; Asa Petersén; Graham S Jackson; Jeremy D Isaacs; Mark Kristiansen; Gillian P Bates; Blair R Leavitt; Geoff Keir; Malcolm Ward; Sarah J Tabrizi Journal: J Proteome Res Date: 2007-06-07 Impact factor: 4.466
Authors: Maria Björkqvist; Edward J Wild; Jenny Thiele; Aurelio Silvestroni; Ralph Andre; Nayana Lahiri; Elsa Raibon; Richard V Lee; Caroline L Benn; Denis Soulet; Anna Magnusson; Ben Woodman; Christian Landles; Mahmoud A Pouladi; Michael R Hayden; Azadeh Khalili-Shirazi; Mark W Lowdell; Patrik Brundin; Gillian P Bates; Blair R Leavitt; Thomas Möller; Sarah J Tabrizi Journal: J Exp Med Date: 2008-07-14 Impact factor: 14.307
Authors: Jennifer C Boatz; Talia Piretra; Alessia Lasorsa; Irina Matlahov; James F Conway; Patrick C A van der Wel Journal: J Mol Biol Date: 2020-06-27 Impact factor: 5.469
Authors: Sonia M Vallabh; Chloe K Nobuhara; Franc Llorens; Inga Zerr; Piero Parchi; Sabina Capellari; Eric Kuhn; Jacob Klickstein; Jiri G Safar; Flavia C Nery; Kathryn J Swoboda; Michael D Geschwind; Henrik Zetterberg; Steven E Arnold; Eric Vallabh Minikel; Stuart L Schreiber Journal: Proc Natl Acad Sci U S A Date: 2019-04-01 Impact factor: 11.205
Authors: Andrew G Hoss; Valentina N Lagomarsino; Samuel Frank; Tiffany C Hadzi; Richard H Myers; Jeanne C Latourelle Journal: Mov Disord Date: 2015-11-17 Impact factor: 10.338