Ultrasensitive and selective detection of 3-repeat tau seeding activity in Pick disease brain and cerebrospinal fluid
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The diagnosis and treatment of diseases involving tau-based pathology such as Alzheimer disease and certain frontotemporal dementias is hampered by the inability to detect pathological forms of tau with sufficient sensitivity, specificity and practicality. In these neurodegenerative diseases, tau accumulates in self-seeding filaments. For example, Pick disease (PiD) is associated with frontotemporal degeneration and accumulation of 3-repeat (3R) tau isoforms in filaments constituting Pick bodies. Exploiting the self-seeding activity of tau deposits, and using a 3R tau fragment as a substrate, we have developed an assay (tau RT-QuIC) that can detect tau seeds in 2 µl aliquots of PiD brain dilutions down to 10−7–10−9. PiD seeding activities were 100-fold higher in frontal and temporal lobes compared to cerebellar cortex. Strikingly, this test was 103- to 105-fold less responsive when seeded with brain containing predominant 4-repeat (4R) tau aggregates from cases of corticobasal degeneration, argyrophilic grain disease, and progressive supranuclear palsy. Alzheimer disease brain, with 3R + 4R tau deposits, also gave much weaker responses than PiD brain. When applied to cerebrospinal fluid samples (5 µl), tau RT-QuIC analyses discriminated PiD from non-PiD cases. These findings demonstrate that abnormal tau aggregates can be detected with high sensitivity and disease-specificity in crude tissue and fluid samples. Accordingly, this tau RT-QuIC assay exemplifies a new approach to diagnosing tauopathies and monitoring therapeutic trials using aggregated tau itself as a biomarker.
KeywordsTau aggregate Pick’s disease Tauopathy RT-QuIC Diagnosis Biomarker Alzheimer's
We thank Lynne Raymond for helpful discussions about protein purification and training in the use of chromatography equipment. We thank David Mead for running MALDI mass spectra for this study and Dr. Vinod Nair for assistance with the electron microscopy. We thank Rose Marie Richardson and Francine Epperson for technical help and research coordination, which has been essential in the process of obtaining and preparing tissue for the best possible diagnoses. We thank Drs. Brent Race and Mr. Andrew Hughson for critical review of this manuscript. This work was supported in part by the Intramural Research Program if the NIAID and the Neuropathology Core, Indiana Alzheimer Disease Center P30 AG010133 and by the Japan Society for the Promotion of Science (JSPS) Fellowship for Japanese Biomedical and Behavioral Researchers at NIH to ES. We are grateful to Dr. Geidy Serrano and the Banner Sun Health Research Institute Brain and Body Donation Program of Sun City, Arizona for the provision of brain tissue and cerebrospinal fluid. The Brain and Body Donation Program is supported by the National Institute on Aging (P30 AG19610 Arizona Alzheimer’s Disease Core Center), the Arizona Biomedical Research Commission (contracts 4001, 0011, 05-901 and 1001 to the Arizona Parkinson’s Disease Consortium) and Prescott Family Initiative of the Michael J. Fox Foundation for Parkinson’s Research.
Coordinated project: ES and BC. Performed experiments: ES (all types), AK (electron microscopy). Provided human brain samples and associated clinical data: BG, AO and GZ. Provided tau KO mouse brain tissue and lysates thereof: JF and MD. Prepared the manuscript: BC and ES. Edited the manuscript: BC, ES, GZ, BG, AK, JF.
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