Ultrasensitive measurement of huntingtin protein in cerebrospinal fluid demonstrates increase with Huntington disease stage and decrease following brain huntingtin suppression

• Published in: Scientific reports Vol. 5; no. 1; p. 12166
• Main Authors: Southwell, Amber L., Smith, Stephen E.P., Davis, Tessa R., Caron, Nicholas S., Villanueva, Erika B., Xie, Yuanyun, Collins, Jennifer A., Li Ye, Min, Sturrock, Aaron, Leavitt, Blair R., Schrum, Adam G., Hayden, Michael R.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 15-07-2015; Nature Publishing Group
• Subjects: 42/89; 631/1647/2017/2215; 631/1647/334/2045; 631/378/1689/1558; 64/110; 82; 82/1; Adult; Aged; Animals; Brain - metabolism; Brain - pathology; Cerebrospinal fluid; Clinical trials; Disease Models, Animal; Female; Flow Cytometry; Gene silencing; Humanities and Social Sciences; Humans; Huntingtin; Huntingtin Protein; Huntington Disease - metabolism; Huntington Disease - pathology; Huntington's disease; Huntingtons disease; Immunoblotting; Immunoprecipitation; Male; Mice; Middle Aged; multidisciplinary; Mutation; Nerve Tissue Proteins - cerebrospinal fluid; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Proteins; Quantitation; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - isolation & purification; Rodents; Science; Severity of Illness Index; Toxicity
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep12166

Quantitation of huntingtin protein in the brain is needed, both as a marker of Huntington disease (HD) progression and for use in clinical gene silencing trials. Measurement of huntingtin in cerebrospinal fluid could be a biomarker of brain huntingtin, but traditional protein quantitation methods have failed to detect huntingtin in cerebrospinal fluid. Using micro-bead based immunoprecipitation and flow cytometry (IP-FCM), we have developed a highly sensitive mutant huntingtin detection assay. The sensitivity of huntingtin IP-FCM enables accurate detection of mutant huntingtin protein in the cerebrospinal fluid of HD patients and model mice, demonstrating that mutant huntingtin levels in cerebrospinal fluid reflect brain levels, increasing with disease stage and decreasing following brain huntingtin suppression. This technique has potential applications as a research tool and as a clinical biomarker.