Ex vivo 18 O-labeling mass spectrometry identifies a peripheral amyloid β clearance pathway

• Published in: Molecular neurodegeneration 2017-02, Vol.12 (1), p.18
• Main Authors: Portelius, Erik, Mattsson, Niklas, Pannee, Josef, Zetterberg, Henrik, Gisslén, Magnus, Vanderstichele, Hugo, Gkanatsiou, Eleni, Crespi, Gabriela A N, Parker, Michael W, Miles, Luke A, Gobom, Johan, Blennow, Kaj
• Format: Article
• Language: English
• Subjects: Alzheimer Disease - cerebrospinal fluid; Alzheimer Disease - metabolism; Amyloid beta-Peptides - cerebrospinal fluid; Amyloid beta-Protein Precursor - cerebrospinal fluid; Antibodies, Monoclonal, Humanized - immunology; Brain - metabolism; Humans; Mass Spectrometry - methods; Oxygen Isotopes; Peptide Fragments - cerebrospinal fluid; Peptide Fragments - metabolism; Proteolysis
• ISSN: 1750-1326

Proteolytic degradation of amyloid β (Aβ) peptides has been intensely studied due to the central role of Aβ in Alzheimer's disease (AD) pathogenesis. While several enzymes have been shown to degrade Aβ peptides, the main pathway of Aβ degradation in vivo is unknown. Cerebrospinal fluid (CSF) Aβ42 is reduced in AD, reflecting aggregation and deposition in the brain, but low CSF Aβ42 is, for unknown reasons, also found in some inflammatory brain disorders such as bacterial meningitis. Using O-labeling mass spectrometry and immune-affinity purification, we examined endogenous proteolytic processing of Aβ in human CSF. The Aβ peptide profile was stable in CSF samples from healthy controls but in CSF samples from patients with bacterial meningitis, showing increased leukocyte cell count, O-labeling mass spectrometry identified proteolytic activities degrading Aβ into several short fragments, including abundant Aβ1-19 and 1-20. After antibiotic treatment, no degradation of Aβ was detected. In vitro experiments located the source of the proteolytic activity to blood components, including leukocytes and erythrocytes, with insulin-degrading enzyme as the likely protease. A recombinant version of the mid-domain anti-Aβ antibody solanezumab was found to inhibit insulin-degrading enzyme-mediated Aβ degradation. O labeling-mass spectrometry can be used to detect endogenous proteolytic activity in human CSF. Using this technique, we found an enzymatic activity that was identified as insulin-degrading enzyme that cleaves Aβ in the mid-domain of the peptide, and could be inhibited by a recombinant version of the mid-domain anti-Aβ antibody solanezumab.