Dataset Information

Plasma Proteome Stability at the Time of Sudden Cardiac Arrest: Implications for Biomarker Studies in Post-Resuscitation Survival

ABSTRACT: Plasma serves as a crucial source of circulating protein biomarkers for advancing resuscitation strategies in sudden cardiac arrest (SCA). However, emergency blood collection under SCA introduces preanalytical variables that may affect proteomic analyses. This study assessed plasma protein stability under blood handling conditions that simulate real-world first responder scenarios. Blood from four individuals was aliquoted and subjected to different (a) initial storage conditions and (b) delays after collection, simulating first responder blood handling scenarios. Plasma was analyzed using data-independent acquisition mass spectrometry (DIA-MS) for global proteomics and parallel reaction monitoring (PRM-MS) for 16 SCA biomarker candidates. Global proteomic variation was primarily driven by inter-individual differences, with less influence from storage conditions or delays. Notably, 87.5% of the candidate biomarkers (14/16) remained stable under ideal storage (4°C storage immediately after blood collection), and 62.5% (10/16) retained stability even under worst-case (room temperature storage for 8 hours after blood collection) for up to 24 hours. However, several proteins lost stability with delayed processing, even under ideal storage. Our study highlights the critical importance of processing blood within 24 hours of collection and accounting for protein stability to preserve the reliability of biomarkers for improving resuscitation strategies in SCA.

ORGANISM(S): Homo Sapiens

SUBMITTER: Jihyeon Lee

PROVIDER: PXD067885 | panorama | Wed Apr 29 00:00:00 BST 2026

REPOSITORIES: PanoramaPublic

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Publications

Plasma Proteome Stability at the Time of Sudden Cardiac Arrest: Implications for Biomarker Studies in Post-Resuscitation Survival.

Lee Jihyeon J Nakamura Kotoka K Fu Qin Q Reinier Kyndaron K Haghani Ali A Chugh Harpriya H Chugh Sumeet S SS Van Eyk Jennifer E JE

Journal of proteome research 20251218 1

Plasma serves as a crucial source of circulating protein biomarkers for advancing resuscitation strategies in sudden cardiac arrest (SCA). However, emergency blood collection under the SCA introduces preanalytical variables that may affect proteomic analyses. This study assessed plasma protein stability under blood-handling conditions that simulate real-world first responder scenarios. Blood from four individuals was aliquoted and subjected to different (a) initial storage conditions and (b) del ...[more]

PMID: 41410264

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Project description:Background: The transcriptome of peripheral white blood cells (PWBCs) contains valuable physiological information, thus making them a prime biological sample for investigating mRNA-based biomarkers. However, prolonged storage of whole blood samples can alter gene transcript abundance in PWBCs, compromising the results of biomarker discovery. Here, we designed an experiment to interrogate the impacts of delayed processing of whole blood samples on gene transcript abundance in PWBCs. We hypothesized that storing blood samples for 24 hours at 4°C would cause RNA degradation resulting in altered transcriptome profiles. Results: We produced RNA-sequencing data for 30 samples collected from five estrus synchronized heifers (Bos taurus). We quantified transcript abundance for 12,414 protein-coding genes in PWBCs. Analysis of parameters of RNA quality revealed no statistically significant differences (P>0.05) between samples collected from the jugular vein and coccygeal vein, as well as among samples processed after 1, 3, 6, or 8 hours. However, samples processed after 24 hours of storage had a significantly lower RNA integrity number value (P=0.03) in comparison to those processed after one hour of storage. Next, we analyzed RNA-sequencing data between samples using those processed after one hour of storage as the baseline for comparison. Interestingly, evaluation of 3’/5’ bias revealed no statistically significant differences between genes with lower transcript abundance in samples stored for 24 hours relative to one hour. In addition, sequencing coverage of transcripts was similar between samples from the 24 hours and one-hour groups. We identified four and 515 genes with differential transcript abundance in samples processed after storage for eight and 24 hours, respectively, relative to samples processed after one hour. Conclusions: The PWBCs respond to prolonged cold storage by increasing genes related to active chromatin compaction which in turn reduces gene transcription. This alteration in transcriptome profiles can impair accuracy of mRNA-based biomarkers. Therefore, blood samples collected for mRNA-based biomarker discovery should be refrigerated immediately and processed within six hours post sampling.

Project description:Metabolomics is a widely used approach for analyzing a vast array of low molecular weight compounds such as amino acids, organic acids, vitamins, biogenic amines and carbohydrates in biological samples, with the aim of investigating biomarkers in personalized medicine studies. Advancements in gas chromatography mass spectrometry (GCMS) instrumentation, along with the availability of commercial and public spectral libraries, have highlighted the relevance of GCMS analysis as a valuable tool for metabolomics applications. Stability assessment in derivatization and GCMS analysis is a crucial yet often overlooked aspect of metabolomics studies. In this study, an untargeted GCMS method workflow for large scale metabolomics studies is presented after assessment and optimization of whole blood sample stability. The method consists of a common two-step derivatization procedure including methoximation using methoxyamine hydrochloride, followed by silylation with N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA). To ensure the stability of the studied metabolites, extensive stability experiments were performed. The stability of the derivatives was evaluated over 24 h in the autosampler at room temperature, as well as after storage for 24 and 48 h at -20 C for both derivatized and dried extracts. While derivatized samples remained stable for 24-48 h in the freezer, dried extracts exhibited variability after 48 h. Findings support the storage of derivatized samples over dried extracts, ensuring greater stability. To increase confidence in metabolite identification, data from the analysis of 120 standard compounds were utilized. The developed method was applied to analyze blood samples from 32 children with ventilator-associated pneumonia (VAP), collected at four different time points during ICU hospitalization. This analysis led to the identification of 43 metabolites. The results of multivariate and univariate statistical analyses demonstrated several statistically significant metabolites, including aspartic acid, alanine, and pyroglutamic acid, which showed a strong correlation with the disease manifestation and may potentially serve as biomarkers in the diagnosis of ventilator-associated pneumonia (VAP) at the stage of clinical suspicion.

Dataset Information

Plasma Proteome Stability at the Time of Sudden Cardiac Arrest: Implications for Biomarker Studies in Post-Resuscitation Survival

Publications

Plasma Proteome Stability at the Time of Sudden Cardiac Arrest: Implications for Biomarker Studies in Post-Resuscitation Survival.

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