Poster abstracts
Poster number 119 submitted by Jacelyn Greenwald
Proteomics of adipose tissue-derived extracellular vesicles from bariatric surgery patients
Jacelyn Greenwald (Neuroscience Graduate Program, Department of Chemistry and Biochemistry, The Ohio State University ), Paola Loreto Palacio (Department of Pediatrics, Division of Neurology, Center for Clinical and Translational Research, Abigail Wexner Research Institute, Nationwide Childrens Hospital), Yongseok Kim (Department of Chemistry and Biochemistry, The Ohio State University ), Setty M. Magaa (Department of Pediatrics, Division of Neurology, Center for Clinical and Translational Research, Abigail Wexner Research Institute, Nationwide Childrens Hospital), Vicki Wysocki (Department of Chemistry and Biochemistry, The Ohio State University )
Abstract:
Mounting evidence supports a relationship between obesity and a nearly twofold increased risk of Alzheimer’s disease (AD).(1,2) However, the cellular and molecular mechanism(s) by which increased adiposity contributes to neurodegeneration are uncharacterized. We are interested in extracellular vesicles (EVs) as key mediators of intercellular crosstalk between adipose tissue and the central nervous system (CNS). EVs are known to cross the blood-brain barrier and to propagate the amyloid beta pathology seen in AD.(3,4) We present the first application of a novel multiparametric pipeline for isolating and characterizing adipose-derived EVs (AdEVs) from tissue.
Microfluidic resistive pulse sensing determined that isolated AdEVs had a concentration of 1.27E+13 particles/mL and a median diameter size of 86.12nm (size range: 69.47 - 140.83nm). Exosome markers (CD63, CD9, CD81) and adipocyte markers (FABP4 and adiponectin) were detected by western blot. Transmission electron microscopy of AdEVs demonstrated characteristic cup-shaped morphology. Bottom-up proteomics data were collected on a Bruker timsTOF Pro instrument and run in parallel accumulation-serial fragmentation (PASEF) mode.
Reactome analysis showed significant proteins implicated in lipid metabolism, providing validation that AdEVs were successfully isolated from tissue. Specifically, 318 entities were found in the AdEV dataset (p-value 1.99E-11 using Binomial Test) and 1,647 total interactions were found. Biological process enrichment of AdEVs revealed highly expressed proteins implicated in cell metabolism, energy pathways, protein metabolism, cell growth and maintenance, transport, immune response, protein folding, and other metabolic, energy and immune response pathways.
Better characterization of adipose tissue-derived EVs may contribute essential information regarding obesity disease status and pave the way toward adipose-enriched protein candidate EV markers in peripheral circulation.
References:
(1) Anstey KJ, Cherbuin N, Budge M, et al. (2011). Body mass index in midlife and late-life as a risk factor for dementia: a meta-analysis of prospective studies. Obesity Reviews, 12(5), e426–e437. DOI: 10.1111/j.1467-789X.2010.00825.x
(2) Picone P, di Carlo M, & Nuzzo D. (2020). Obesity and Alzheimer’s disease: Molecular bases. European J Neurosci, 52(8), 3944–3950. DOI: 10.1111/ejn.14758
(3) Morad G, Carman CV, Hagedorn EJ, et al. (2019). Tumor-Derived Extracellular Vesicles Breach the Intact Blood–Brain Barrier via Transcytosis. ACS Nano, 13(12), 13853–13865. DOI: 10.1021/acsnano.9b04397
(4) Gabrielli M, Prada I, Joshi P, et al. (2022). Microglial large extracellular vesicles propagate early synaptic dysfunction in Alzheimer’s disease. Brain, 145(8), 2849–2868. DOI: 10.1093/brain/awac083
Keywords: Extracellular vesicles, proteomics, mass spectrometry