Barocycler-Based Concurrent Multiomics Method To Assess Molecular Changes Associated with Atherosclerosis Using Small Amounts of Arterial Tissue from a Single Mouse
datasetposted on 24.09.2019, 20:29 by Jihan Talib, Peter G. Hains, Sergey Tumanov, Mark P. Hodson, Phillip J. Robinson, Roland Stocker
Atherosclerosis is a complex, multifactorial disease characterized by the buildup of plaque in the arterial wall. Apolipoprotein E gene deficient (Apoe–/–) mice serve as a commonly used tool to elucidate the pathophysiology of atherosclerosis because of their propensity to spontaneously develop arterial lesions. To date, however, an integrated omics assessment of atherosclerotic lesions in individual Apoe–/– mice has been challenging because of the small amount of diseased and nondiseased tissue available. To address this current limitation, we developed a multiomics method (Multi-ABLE) based on the proteomic method called accelerated Barocycler lysis and extraction (ABLE) to assess the depth of information that can be obtained from arterial tissue derived from a single mouse by splitting ABLE to allow for a combined proteomics–metabolomics–lipidomics analysis (Multi-ABLE). The new method includes tissue lysis via pressure cycling technology (PCT) in a Barocycler, followed by proteomic analysis of half the sample by nanoLC-MS and sequential extraction of lipids (organic extract) and metabolites (aqueous extract) combined with HILIC and reversed phase chromatography and time-of-flight mass spectrometry on the other half. Proteomic analysis identified 845 proteins, 93 of which were significantly altered in lesion-containing arteries. Lipidomic and metabolomic analyses detected 851 lipid and 362 metabolite features, which included 215 and 65 identified lipids and metabolites, respectively. The Multi-ABLE method is the first to apply a concurrent multiomics pipeline to cardiovascular disease using small (<5 mg) tissue samples, and it is applicable to other diseases where limited size samples are available at specific points during disease progression.
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Apoepressure cycling technologymultifactorial diseaseArterial TissueMultiomics Methodmetabolomic analysesSingle Mouse Atherosclerosis845 proteinshalfProteomic analysistime-of-flight mass spectrometrylesion-containing arteriesApolipoprotein E geneproteomic analysisPCTMolecular Changes AssociatedMulti-ABLE methodSmall Amountsmultiomics methodomics assessmentphase chromatography362 metabolite features851 lipidnondiseased tissuesize samplestissue samplesatherosclerotic lesionsmultiomics pipelineHILICsequential extractionsplitting ABLEtissue lysisdisease progressionBarocycler lysisproteomic method