title: Microwave and magnetic (M(2) ) proteomics of the experimental autoimmune encephalomyelitis animal model of multiple sclerosis.
abstract: We hypothesized that quantitative MS/MS-based proteomics at multiple time points, incorporating rapid microwave and magnetic (M(2) ) sample preparation, could enable relative protein expression to be correlated to disease progression in the experimental autoimmune encephalomyelitis (EAE) animal model of multiple sclerosis. To test our hypothesis, microwave-assisted reduction/alkylation/digestion of proteins from brain tissue lysates bound to C8 magnetic beads and microwave-assisted isobaric chemical labeling were performed of released peptides, in 90 s prior to unbiased proteomic analysis. Disease progression in EAE was assessed by scoring clinical EAE disease severity and confirmed by histopathologic evaluation for central nervous system inflammation. Decoding the expression of 283 top-ranked proteins (p <0.05) at each time point relative to their expression at the peak of disease, from a total of 1191 proteins observed in four technical replicates, revealed a strong statistical correlation to EAE disease score, particularly for the following four proteins that closely mirror disease progression: 14-3-3ε (p = 3.4E-6); GPI (p = 2.1E-5); PLP1 (p = 8.0E-4); PRX1 (p = 1.7E-4). These results were confirmed by Western blotting, signaling pathway analysis, and hierarchical clustering of EAE risk groups. While validation in a larger cohort is underway, we conclude that M(2) proteomics is a rapid method to quantify putative prognostic/predictive protein biomarkers and therapeutic targets of disease progression in the EAE animal model of multiple sclerosis.
authors.affiliation: Department of Biology, University of Texas at San Antonio, San Antonio, TX, USA.
authors.forename: Itay
authors.initials: I
authors.lastname: Raphael
authors.forename: Swetha
authors.initials: S
authors.lastname: Mahesula
authors.forename: Karan
authors.initials: K
authors.lastname: Kalsaria
authors.forename: Venkat
authors.initials: V
authors.lastname: Kotagiri
authors.forename: Anjali B
authors.initials: AB
authors.lastname: Purkar
authors.forename: Manjushree
authors.initials: M
authors.lastname: Anjanappa
authors.forename: Darshit
authors.initials: D
authors.lastname: Shah
authors.forename: Vidya
authors.initials: V
authors.lastname: Pericherla
authors.forename: Yeshwant Lal Avinash
authors.initials: YL
authors.lastname: Jadhav
authors.forename: Rekha
authors.initials: R
authors.lastname: Raghunathan
authors.forename: Michael
authors.initials: M
authors.lastname: Vaynberg
authors.forename: David
authors.initials: D
authors.lastname: Noriega
authors.forename: Nazul H
authors.initials: NH
authors.lastname: Grimaldo
authors.forename: Carola
authors.initials: C
authors.lastname: Wenk
authors.forename: Jonathan A L
authors.initials: JA
authors.lastname: Gelfond
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Forsthuber
authors.forename: William E
authors.initials: WE
authors.lastname: Haskins
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism / Q000503:physiopathology; D001923:Brain Chemistry; D016000:Cluster Analysis; D004195:Disease Models, Animal; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000378:metabolism*; D005260:Female; D008280:Magnetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008872:Microwaves; D009103:Multiple Sclerosis / Q000378:metabolism; D020543:Proteome / Q000032:analysis / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D020543:Proteome