Blood neuro-exosomal synaptic proteins predict Alzheimer's disease at the asymptomatic stage

Longfei Jia, Min Zhu, Chaojun Kong, Yana Pang, Heng Zhang, Qiongqiong Qiu, Cuibai Wei, Yi Tang, Qi Wang, Ying Li, Tingting Li, Fangyu Li, Qigeng Wang, Yan Li, Yiping Wei, Jianping Jia, Longfei Jia, Min Zhu, Chaojun Kong, Yana Pang, Heng Zhang, Qiongqiong Qiu, Cuibai Wei, Yi Tang, Qi Wang, Ying Li, Tingting Li, Fangyu Li, Qigeng Wang, Yan Li, Yiping Wei, Jianping Jia

Abstract

Introduction: Exosomes are an emerging candidate for biomarkers of Alzheimer's disease (AD). This study investigated whether exosomal synaptic proteins can predict AD at the asymptomatic stage.

Methods: We conducted a two-stage-sectional study (discovery stage: AD, 28; amnestic mild cognitive impairment [aMCI], 25; controls, 29; validation stage: AD, 73; aMCI, 71; controls, 72), a study including preclinical AD (160) and controls (160), and a confirmation study in familial AD (mutation carriers: 59; non-mutation carriers: 62).

Results: The concentrations of growth associated protein 43 (GAP43), neurogranin, synaptosome associated protein 25 (SNAP25), and synaptotagmin 1 were lower in AD than in controls (P < .001). Exosomal biomarker levels were correlated with those in cerebrospinal fluid (R2 = 0.54-0.70). The combination of exosomal biomarkers detected AD 5 to 7 years before cognitive impairment (area under the curve = 0.87-0.89).

Discussion: This study revealed that exosomal GAP43, neurogranin, SNAP25, and synaptotagmin 1 act as effective biomarkers for prediction of AD 5 to 7 years before cognitive impairment.

Keywords: Alzheimer's disease; biomarker; diagnosis; exosome; prediction; synaptic protein.

Conflict of interest statement

The authors declare that they have no conflicts of interest.

© 2020 The Authors. Alzheimer's & Dementia published by Wiley Periodicals, Inc. on behalf of Alzheimer's Association.

Figures

FIGURE 1
FIGURE 1
Four datasets were included in this study. A, Numbers of subjects, and measurements in four datasets. B, Cognitively normal subjects that were included from a longitudinal study between 2012 and 2014. The subjects were grouped into preclinical Alzheimer's disease and controls and followed up in the present study by measuring their cognitive function, and amyloid beta 42, P‐tau, and T‐tau in cerebrospinal fluid. The exosomes were extracted from the blood collected at baseline
FIGURE 2
FIGURE 2
Biomarkers were measured in the discovery and validation datasets. Panels A—D show levels of neuronal‐derived exosomal growth associated protein 43 (GAP43; A), neurogranin (B), synaptosome associated protein 25 (SNAP25; C), and synaptotagmin 1 (D). Panels E–H show levels of cerebrospinal fluid (CSF) GAP43 (E), neurogranin (F), SNAP25 (G), and synaptotagmin 1 (H). In the discovery stage, n = 28 (Alzheimer's disease [AD]), 25 (amnestic mild cognitive impairment [aMCI]), and 29 (controls). In the validation stage, n = 73 (AD), 71 (aMCI), and 72 (controls). Abbreviation: Con,  controls *** P < 0.001, ** P < 0.01, * P < 0.05
FIGURE 3
FIGURE 3
The concentrations of exosomal biomarkers are highly correlated with those in the cerebrospinal fluid (CSF). Panels A–C show that levels of growth associated protein 43 in exosomes and the CSF were closely correlated in patients with Alzheimer's disease (AD, A), patients with amnestic mild cognitive impairment (aMCI, B), and controls (C). Panels D–F show robust correlations between neurogranin levels in exosomes and those in the CSF of patients with AD (D), patients with aMCI (E), and controls (F). Panels G–I show significant correlations between synaptosome associated protein 25 (SNAP25) expression in exosomes and those in the CSF in patients with AD (G), patients with aMCI (H), and controls (I). Panels J–L show high correlations between synaptotagmin 1 levels in exosomes and the CSF of patients with AD (J), patients with aMCI (K), and controls (L). The blue circles and lines correspond to the discovery dataset, while the green circles and lines correspond to the validation dataset. In the discovery stage, n = 28 (AD), 25 (aMCI), and 29 (controls). In the validation stage, n = 73 (AD), 71 (aMCI), and 72 (controls)
FIGURE 4
FIGURE 4
Prediction of exosomal growth associated protein 43 (GAP43), neurogranin, synaptosome associated protein 25 (SNAP25), and synaptotagmin 1 for preclinical Alzheimer's disease (AD). Panels A–D show levels of neuronal‐derived exosomal GAP43 (A), neurogranin (B), SNAP25 (C), and synaptotagmin 1 (D) in preclinical AD and controls. Panels E–H show receiver operating characteristic (ROC) analyses of GAP43 (E), neurogranin (F), SNAP25 (G), and synaptotagmin 1 (H). Panels I–K show ROC analyses by combining exosomal GAP43, neurogranin, SNAP25, synaptotagmin 1, and APOE status in total dataset (I), randomly selected training dataset (J), and test dataset (K). In addition, the ROC of APOE ε4 status was analyzed independently in total dataset (L). n = 160 (preclinical AD) and 160 (controls). Pre‐AD = preclinical AD
FIGURE 5
FIGURE 5
Prediction of exosomal growth associated protein 43 (GAP43), neurogranin, synaptosome associated protein 25 (SNAP25), and synaptotagmin 1 for mutation carriers in a familial Alzheimer's disease (FAD) cohort (dataset 4). Panels A–D show levels of neuronal‐derived exosomal GAP43 (A), neurogranin (B), SNAP25 (C), and synaptotagmin 1 (D) in mutation carriers and non‐mutation controls. Panels E–H show ROC analyses of GAP43 (E), neurogranin (F), SNAP25 (G), and synaptotagmin 1 (H). Panel I shows receiver operating characteristic (ROC) analyses by applying the predict model generated from preclinical AD dataset (dataset 3). n = 59 (mutation carriers) and 62 (non‐mutation carriers). Abbreviations: Con,  controls, Mut,  mutation carriers

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