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Type: Journal article
Title: Proteomic analysis of human plasma during intermittent fasting
Author: Harney, D.J.
Hutchison, A.T.
Hatchwell, L.
Humphrey, S.J.
James, D.E.
Hocking, S.
Heilbronn, L.K.
Larance, M.
Citation: Journal of Proteome Research, 2019; 18(5):2228-2240
Publisher: American Chemical Society
Issue Date: 2019
ISSN: 1535-3893
Statement of
Dylan J. Harney, Amy T. Hutchison, Luke Hatchwell, Sean J. Humphrey, David E. James, Samantha Hocking, Leonie K. Heilbronn and Mark Larance
Abstract: Intermittent fasting (IF) increases lifespan and decreases metabolic disease phenotypes and cancer risk in model organisms, but the health benefits of IF in humans are less clear. Human plasma derived from clinical trials is one of the most difficult sample sets to analyze using mass spectrometry-based proteomics due to the extensive sample preparation required and the need to process many samples to achieve statistical significance. Here, we describe an optimized and accessible device (Spin96) to accommodate up to 96 StageTips, a widely used sample preparation medium enabling efficient and consistent processing of samples prior to LC-MS/MS. We have applied this device to the analysis of human plasma from a clinical trial of IF. In this longitudinal study employing 8-weeks IF, we identified significant abundance differences induced by the IF intervention, including increased apolipoprotein A4 (APOA4) and decreased apolipoprotein C2 (APOC2) and C3 (APOC3). These changes correlated with a significant decrease in plasma triglycerides after the IF intervention. Given that these proteins have a role in regulating apolipoprotein particle metabolism, we propose that IF had a positive effect on lipid metabolism through modulation of HDL particle size and function. In addition, we applied a novel human protein variant database to detect common protein variants across the participants. We show that consistent detection of clinically relevant peptides derived from both alleles of many proteins is possible, including some that are associated with human metabolic phenotypes. Together, these findings illustrate the power of accessible workflows for proteomics analysis of clinical samples to yield significant biological insight.
Keywords: 3D-printing; 96-well; human; intermittent fasting; liquid chromatography; mass spectrometry (MS); plasma; sample cleanup; solid-phase extraction (SPE)
Rights: © 2019 American Chemical Society
RMID: 0030112664
DOI: 10.1021/acs.jproteome.9b00090
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Appears in Collections:Medicine publications

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