N-Terminally Truncated and Pyroglutamate-Modified Aβ Forms Are Measurable in Human Cerebrospinal Fluid and Are Potential Markers of Disease Progression in Alzheimer's Disease.

Abstract

Alzheimer's disease (AD) is a pathology characterized by the accumulation in the brain of intracellular and extracellular amyloid-β (Aβ) aggregates, especially of Aβ1-40 and Aβ1-42 peptides. It is known that N-terminally truncated or modified Aβ forms also exist in AD brains and cerebrospinal fluid (CSF), and they play a key role in the pathogenesis of the disease. Herein, we developed an antibody-free method based on Solid-Phase Extraction and Electrospray Ionization Liquid Chromatography Mass Spectrometry for the identification and quantitation in human CSF of Aβ isoforms. In human CSF, we could detect and quantify a panel of 19 Aβ isoforms, including N-terminally truncated and pyroglutamate-modified forms, never quantified before in CSF. Among these, we identified novel N-terminally truncated Aβ species: four bound to copper and two phosphorylated forms, which were found to be the most common proteoforms in human CSF along with Aβ1-40, Aβ3-40, and AβpE11-42. We tested the newly developed and validated method in a pilot study on CSF from elderly individuals with subjective memory complaints (SMCs, n = 9), mild cognitive impairment (MCI, n = 18), and AD (n = 15); along with Aβ1-42, five N-terminally truncated forms (Aβ11-40, Aβ3-42, AβpE11-42, AβpE3-40, and Aβ4-40 Cu2+) are altered in AD/MCI. Thus, we demonstrated that N-terminally truncated and pyroglutamate-modified Aβ can be quantified in human CSF, and five of them, along with Aβ1-42, are potential markers of AD progression. The described method could represent a useful tool for patients' stratification and monitoring. Moreover, the newly identified Aβ CSF species might represent new potential therapeutic targets.

• Authors: Guido Domingo, Luisa Benussi, Claudia Saraceno, Michela Bertuzzi, Roland Nicsanu, Antonio Longobardi, Sonia Bellini, Alfredo Cagnotto, Mario Salmona, Giuliano Binetti, Roberta Ghidoni
• Journal: Frontiers in neuroscience (Front Neurosci) Vol. 15 Pg. 708119 ( 2021) ISSN: 1662-4548 [Print] Switzerland
• PMID: 34393717 (Publication Type: Journal Article)
• Copyright: Copyright © 2021 Domingo, Benussi, Saraceno, Bertuzzi, Nicsanu, Longobardi, Bellini, Cagnotto, Salmona, Binetti and Ghidoni.