Changes in Synaptic Proteins Precede Neurodegeneration Markers in Preclinical Alzheimer's Disease Cerebrospinal Fluid

• Published in: Molecular & cellular proteomics Vol. 18; no. 3; pp. 546 - 560
• Main Authors: Lleó, Alberto, Núñez-Llaves, Raúl, Alcolea, Daniel, Chiva, Cristina, Balateu-Paños, Daniel, Colom-Cadena, Martí, Gomez-Giro, Gemma, Muñoz, Laia, Querol-Vilaseca, Marta, Pegueroles, Jordi, Rami, Lorena, Lladó, Albert, Molinuevo, José L., Tainta, Mikel, Clarimón, Jordi, Spires-Jones, Tara, Blesa, Rafael, Fortea, Juan, Martínez-Lage, Pablo, Sánchez-Valle, Raquel, Sabidó, Eduard, Bayés, Àlex, Belbin, Olivia
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.03.2019; The American Society for Biochemistry and Molecular Biology
• Subjects: Aged; Alzheimer Disease - cerebrospinal fluid; Alzheimer Disease - metabolism; Alzheimer's disease; Autopsy; Biomarker: Prognostic; Biomarkers - cerebrospinal fluid; Biomarkers - metabolism; Calcium-Binding Proteins - cerebrospinal fluid; Calcium-Binding Proteins - metabolism; Cerebrospinal fluid; Clinical proteomics; Early Diagnosis; Female; Humans; Male; Nerve Tissue Proteins - cerebrospinal fluid; Nerve Tissue Proteins - metabolism; Prodromal Symptoms; Prognosis; Proteomics - methods; Receptors, AMPA - metabolism; Selected reaction monitoring; Synapses - metabolism; Syntaxin 1 - cerebrospinal fluid; Syntaxin 1 - metabolism; Thy-1 Antigens - cerebrospinal fluid; Thy-1 Antigens - metabolism; Biomarker: Prognostic; Cerebrospinal fluid; Selected reaction monitoring; Clinical proteomics; Alzheimer's disease
• ISSN: 1535-9476; 1535-9484; 1535-9484
• DOI: 10.1074/mcp.RA118.001290

In the search for biomarkers of synapse loss associated with Alzheimer's disease (AD), we used shotgun proteomics to identify a panel of 9 synaptic proteins detectable in human cerebrospinal fluid (CSF). Expression at the human synapse was verified using super resolution microscopy. Using SRM, we monitored the panel in CSF from 3 independent clinical cohorts. We report 6 synaptic biomarkers that demonstrate changes in preclinical AD prior to markers of neurodegeneration, which could have clinical value for assessing disease progression. [Display omitted] Highlights •Proteomic analysis of cerebrospinal fluid and identification of synaptic component.•Use of super resolution microscopy to verify synapse-specificity in human tissue.•Selective reaction monitoring MS (SRM) of synaptic panel in 3 cohorts of Alzheimer's disease cerebrospinal fluid.•Synaptic protein changes precede tau in preclinical Alzheimer's disease. A biomarker of synapse loss, an early event in Alzheimer's disease (AD) pathophysiology that precedes neuronal death and symptom onset, would be a much-needed prognostic biomarker. With direct access to the brain interstitial fluid, the cerebrospinal fluid (CSF) is a potential source of synapse-derived proteins. In this study, we aimed to identify and validate novel CSF biomarkers of synapse loss in AD. Discovery: Combining shotgun proteomics of the CSF with an exhaustive search of the literature and public databases, we identified 251 synaptic proteins, from which we selected 22 for further study. Verification: Twelve proteins were discarded because of poor detection by Selected Reaction Monitoring (SRM). We confirmed the specific expression of 9 of the remaining proteins (Calsyntenin-1, GluR2, GluR4, Neurexin-2A, Neurexin-3A, Neuroligin-2, Syntaxin-1B, Thy-1, Vamp-2) at the human synapse using Array Tomography microscopy and biochemical fractionation methods. Exploration: Using SRM, we monitored these 9 synaptic proteins (20 peptides) in a cohort of CSF from cognitively normal controls and subjects in the pre-clinical and clinical AD stages (n = 80). Compared with controls, peptides from 8 proteins were elevated 1.3 to 1.6-fold (p < 0.04) in prodromal AD patients. Validation: Elevated levels of a GluR4 peptide at the prodromal stage were replicated (1.3-fold, p = 0.04) in an independent cohort (n = 60). Moreover, 7 proteins were reduced at preclinical stage 1 (0.6 to 0.8-fold, p < 0.04), a finding that was replicated (0.7 to 0.8-fold, p < 0.05) for 6 proteins in a third cohort (n = 38). In a cross-cohort meta-analysis, 6 synaptic proteins (Calsyntenin-1, GluR4, Neurexin-2A, Neurexin-3A, Syntaxin-1B and Thy-1) were reduced 0.8-fold (p < 0.05) in preclinical AD, changes that precede clinical symptoms and CSF markers of neurodegeneration. Therefore, these proteins could have clinical value for assessing disease progression, especially in preclinical stages of AD.