Abstract
BackgroundProteomic technologies applied for profiling human biofluids and blood cells are considered to reveal new biomarkers of exposure or provide insights into novel mechanisms of adaptation.MethodsBoth a non-targeted (classical 2D-electrophoresis combined with mass spectrometry) as well as a targeted proteomic approach (multiplex immunoassay) were applied to investigate how fasting for 36 h, as compared to 12 h, affects the proteome of platelets, peripheral blood mononuclear cells (PBMC), plasma, urine and saliva collected from ten healthy volunteers.ResultsBetween-subject variability was highest in the plasma proteome and lowest in the PBMC proteome. Random Forests analysis performed on the entire dataset revealed that changes in the level of the RhoGDI2 protein in PBMC and plasma ApoA4 levels were the two most obvious biomarkers of an extended fasting. Random Forests (RF) analysis of the multiplex immunoassay data revealed leptin and MMP-3 as biomarkers for extended fasting. However, high between-subject variability may have masked the extended fasting effects in the proteome of the biofluids and blood cells.ConclusionsIdentification of significantly changed proteins in biofluids and blood cells using a non-targeted approach, together with the outcome of targeted analysis revealed both known and novel markers for a 36 h fasting period, including the cellular proteins RhoGDI2 and CLIC1, and plasma proteins ApoA4, leptin and MMP-3. The PBMC proteome exhibited the lowest between-subject variability and therefore these cells appear to represent the best biosamples for biomarker discovery in human nutrigenomics.
Highlights
Proteomic technologies applied for profiling human biofluids and blood cells are considered to reveal new biomarkers of exposure or provide insights into novel mechanisms of adaptation
In order not to miss any interesting spots we performed additional analyses on the original scale and selected proteins with an adjusted P below 0.2 on this scale. In this way we identified 10 proteins from the 2D-electrophoresis and 39 proteins for the multiplex immunoassay which were significantly changed upon the extended fasting
For example, extended fasting resulted in a significant alteration of only 1.3% of the protein spots, whereas 69.1% of the spots differed significantly between subjects, when using 2D-electrophoresis
Summary
Proteomic technologies applied for profiling human biofluids and blood cells are considered to reveal new biomarkers of exposure or provide insights into novel mechanisms of adaptation. Proteomic technologies are increasingly being applied in nutrition research to reveal biomarkers that can help to demonstrate effectiveness of certain diets. These technologies aid the discovery of mechanisms whereby dietary regimens fluid or blood cells would be most appropriate to reflect such responses. This human trial was part of the European Nutrigenomics Organization (NuGO) Proof of Principal Study (PPS) in which various omic techniques were applied [7], including transcriptomics, proteomics and metabolomics [8]. This component of the PPS was designed to evaluate experimental and biological variation on the individual level in nutrigenomic experiments as a basis for future personalized nutrition concepts [9]
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