Nascent Proteomes in Peripheral Blood Mononuclear Cells as a Novel Source for Biomarker Discovery in Human Stroke

Abstract

Background and Purpose The proteome of newly synthesized proteins (nascent proteome) in peripheral blood mononuclear cells (PBMC) can be a novel source of stroke biomarkers. Changes in the PBMC nascent proteome after stroke reflect the dynamic response-in-action not detectable in the total proteome (all existing proteins) in blood. Here we test the application of nascent proteomics as a novel approach for stroke biomarker discovery. Methods The PBMC nascent proteome in human blood was determined by metabolic labeling of fresh PBMC cultures with azidohomoalanine (AHA, an azide-containing methionine surrogate), followed by mass spectrometry detection and quantitation of AHA-labeled proteins. The PBMC nascent and total proteomes were compared between stroke patients and matched controls. Results Both PBMC nascent and total proteomes showed differences between stroke patients and controls. Results of hierarchical clustering analysis of proteomic data revealed greater changes in the nascent than in the total PBMC proteomes, supporting the usefulness of the PBMC nascent proteome as a novel source of stroke biomarkers. Conclusions Nascent proteomes in PBMC can be a novel source for biomarker discovery in human stroke.

A New Strategy to Uncover Fragile X Proteomic Biomarkers Using the Nascent Proteome of Peripheral Blood Mononuclear Cells (PBMCs)

“…The two hours timeframe used to label nascent proteins may also have contributed to reduce the number of proteins identi ed since it favors only proteins with high turnover rate. Despites these potential limitations, the number of nascent proteins identi ed in PBMCs is similar to the one reported by another study [69]. Our experimental work ow, which consisted in the processing of pooled samples, can also have limited the scope of our analysis by limiting our capacity to individually assess each participant's unique proteome.…”

Blood transcriptome changes after stroke in an African American population

“…For example, efficacy (surrogate) biomarkers, such as beta amyloid have been investigated for Alzheimer's disease therapies, and the release of intracellular proteins from neurons has been a focus as a biomarker for brain injury 21. While proteomic approaches to identify panels of proteins are becoming more common,22 few single protein biomarkers have the sophistication to subtype neurological injuries. As such the application of gene expression analysis, using unbiased application of mathematical cluster analysis to identify stroke subtypes offers promise.…”

A reference library of peripheral blood mononuclear cells for SWATH‐MS analysis

“…Moreover, PBMCs can be easily and quickly isolated from whole blood through a low-invasive method of collection (i.e., blood drawing) without the interference of highly abundant plasma proteins [2]. For these reasons, PBMCs have been used in proteomic approaches for biomarkers discovery of the most diverse diseases such as stroke [3], amyotrophic lateral sclerosis [4], and schizophrenia [5].…”