Increased Levels of Extracellular Microvesicle Markers and Decreased Levels of Endocytic/Exocytic Proteins in the Alzheimer’s Disease Brain

Article type: Research Article

Authors: Musunuri, Sravani^{a; 1} | Khoonsari, Payam Emami^{b; 1} | Mikus, Maria^c | Wetterhall, Magnus^e | Häggmark-Mänberg, Anna^c | Lannfelt, Lars^d | Erlandsson, Anna^d | Bergquist, Jonas^a | Ingelsson, Martin^d | Shevchenko, Ganna^a | Nilsson, Peter^c | Kultima, Kim^{b; *}

Affiliations: [a] Analytical Chemistry, Department of Chemistry-BMC, Uppsala University, Uppsala, Sweden | [b] Department of Medical Sciences, Cancer Pharmacology and Computational Medicine, Uppsala University Academic Hospital, Uppsala, Sweden | [c] Affinity Proteomics, Science for Life Laboratory, School of Biotechnology, KTH Royal Institute of Technology, Stockholm, Sweden | [d] Department of Public Health/ Geriatrics, Uppsala University, Uppsala, Sweden | [e] GE Healthcare Life Sciences, Uppsala, Sweden

Correspondence: [*] Correspondence to: Dr. Kim Kultima, Department of Medical Sciences, Cancer Pharmacology and Computational Medicine, Uppsala University Academic Hospital, Uppsala, Sweden. Tel.: +46 18 611 4248; Fax: +46 18 611 3703; E-mail: kim.kultima@medsci.uu.se.

Note: [1] These authors contributed equally to this work.

Abstract: Background: Alzheimer’s disease (AD) is a chronic neurodegenerative disorder accounting for more than 50% of all dementia cases. AD neuropathology is characterized by the formation of extracellular plaques and intracellular neurofibrillary tangles consisting of aggregated amyloid-β and tau, respectively. The disease mechanism has only been partially elucidated and is believed to also involve many other proteins. Objective: This study intended to perform a proteomic profiling of post mortem AD brains and compare it with control brains as well as brains from other neurological diseases to gain insight into the disease pathology. Methods: Here we used label-free shotgun mass spectrometry to analyze temporal neocortex samples from AD, other neurological disorders, and non-demented controls, in order to identify additional proteins that are altered in AD. The mass spectrometry results were verified by antibody suspension bead arrays. Results: We found 50 proteins with altered levels between AD and control brains. The majority of these proteins were found at lower levels in AD. Pathway analyses revealed that several of the decreased proteins play a role in exocytic and endocytic pathways, whereas several of the increased proteins are related to extracellular vesicles. Using antibody-based analysis, we verified the mass spectrometry results for five representative proteins from this group of proteins (CD9, HSP72, PI42A, TALDO, and VAMP2) and GFAP, a marker for neuroinflammation. Conclusions: Several proteins involved in exo-endocytic pathways and extracellular vesicle functions display altered levels in the AD brain. We hypothesize that such changes may result in disturbed cellular clearance and a perturbed cell-to-cell communication that may contribute to neuronal dysfunction and cell death in AD.

Keywords: Alzheimer’s disease, endocytosis, exocytosis, exosomes, extracellular vesicles, immobilized, chemistry antibodies, mass spectrometry

DOI: 10.3233/JAD-160271

Journal: Journal of Alzheimer's Disease, vol. 54, no. 4, pp. 1671-1686, 2016