AUTHOR=Cohn Whitaker , Melnik Mikhail , Huang Calvin , Teter Bruce , Chandra Sujyoti , Zhu Chunni , McIntire Laura Beth , John Varghese , Gylys Karen H. , Bilousova Tina 

TITLE=Multi-Omics Analysis of Microglial Extracellular Vesicles From Human Alzheimer’s Disease Brain Tissue Reveals Disease-Associated Signatures

JOURNAL=Frontiers in Pharmacology

VOLUME=Volume 12 - 2021

YEAR=2021

URL=https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2021.766082

DOI=10.3389/fphar.2021.766082

ISSN=1663-9812

ABSTRACT=AAlzheimer’s Disease (AD) is the most common cause of dementia, yet there is no cure nor diagnostics available prior to the onset of clinical symptoms.  Extracellular vesicles (EVs) are lipid bilayer-delimited particles that are released from almost all types of cell. Genome-wide association studies have linked multiple AD genetic risk factors to microglia-specific pathways.  It is plausible that microglia-derived EVs may play a role in the progression of AD by contributing to the dissemination of insoluble pathogenic proteins, such as tau and Aβ. Despite the potential utility of EVs as a diagnostic tool, our knowledge of human brain EV subpopulations is limited. Here we present a method for isolating microglial CD11b-positive small EVs from cryopreserved human brain tissue, as well as an integrated multi-omics analysis of microglial EVs enriched from the parietal cortex of 4 late-stage AD (Braak V-VI) and 3 age-matched normal/low pathology (NL) cases. This integrated analysis revealed 1000 proteins, 594 lipids and 105 miRNAs using shotgun proteomics, targeted lipidomics and NanoString nCounter technology, respectively. The results showed a significant reduction in the abundance of homeostatic microglia markers P2RY12 and TMEM119, and increased levels of disease-associated microglia markers FTH1 and TREM2, in CD11b-positive EVs from AD brain compared to NL cases. Tau abundance was significantly higher in AD brain-derived microglial EVs. These changes were accompanied by the upregulation of synaptic and neuron-specific proteins in the AD group. Levels of free-cholesterol were elevated in microglial EVs from AD brain. Lipidomic analysis also revealed a pro-inflammatory lipid profile, endo-lysosomal dysfunction and a significant AD-associated decrease in levels of docosahexaenoic acid (DHA)-containing polyunsaturated lipids, suggesting a potential defect in acyl-chain remodeling. Additionally, 4 miRNAs associated with immune and cellular senescence signaling pathways were significantly upregulated in the AD group. Our data suggest that loss of the homeostatic microglia signature in late AD stages may be accompanied by endo-lysosomal impairment and the release of undigested neuronal and myelin debris, including tau, through extracellular vesicles. We suggest the analysis of microglia-derived EVs has merit for identifying novel EV-associated biomarkers and providing a framework for future larger-scale multi-omics studies on patient-derived cell-type specific EVs.