MSACL 2025 Abstract

Population-based Proteomics: Platelet Data for Elucidating Mechanisms of Cardiovascular Diseases

Yu-Heng Hsieh, Fiorella A. Solari and Albert Sickmann
Leibniz-Institut für Analytische Wissenschaften – ISAS – e.V., Dortmund, Germany

Albert Sickmann (Presenter) ISAS

Relevant Financial Disclosures (within past 24 months, reported on Mar 28, 2025) No relevant financial relationship(s) to disclose.

INTRODUCTION:
Platelets are small, anucleated cell fragments circulating in the bloodstream, playing a crucial role in thrombus formation following endothelial injury. Dysfunction in platelet mechanisms can contribute to the onset of cardiovascular diseases (CVDs), which are the leading cause of mortality worldwide, responsible for an estimated 17.9 million deaths annually (1). This category includes coronary heart disease, cerebrovascular disease, lung embolism and stroke, among others.

OBJECTIVE:
The primary objective is to generate large-scale platelet population data to observe protein abundance differences in basal platelet states across different age and gender groups. This foundational platelet proteome data, in combination with computational modeling strategies, will aid in developing a highly relevant platelet model. This model will facilitate the analysis of patient samples with CVDs, identify biomarkers for stroke risk and bleeding disorders, and predict drug responses in patients.

METHODS:
To achieve these aims, state-of-the-art mass spectrometry-based proteomics approaches have been employed, integrated with computational modeling strategies. A positive-pressure Filter Aided Sample Preparation (FASP) in a 96-well format (PF96) was developed (2), enabling highly reproducible, time- and cost-efficient analysis of large sample cohorts. This method is combined with robust Liquid Chromatography coupled online to a Mass Spectrometer, utilizing Data-Independent Acquisition (DIA) for enhanced coverage and reproducibility.

RESULTS:
Consequently, these approaches allow for the scale-up of sample input, facilitating population-based studies on platelet biology, extending beyond the analysis of just 4 donors to over 2000, thereby elucidating the ground-truth platelet proteome data.
Analysis of 1500 platelet samples from healthy volunteers aged 18 to 68 years quantified approximately 4,000 platelet proteins in all samples. Preliminary data from the first samples revealed proteomic differences between gender and age groups for proteins such as Purinergic receptor (P2RY12), inositol 1,4,5-triphosphate receptor-associated 2 (IRAG2), vasodilator-stimulated phosphoprotein (VASP), Glycoprotein VI (GP6), and Fibrinogen.

CONCLUSION:
This approach aims to provide insights into malfunctioning platelet mechanisms, offering opportunities for faster diagnostic and therapeutic strategies. Additionally, mapping platelet molecular mechanisms may serve as a blueprint for studying other blood cells, such as neutrophils involved in cancer development and erythrocytes.

REFERENCES:
1. Burkhart JM, Gambaryan S, Watson SP, Jurk K, Walter U, Sickmann A, Heemskerk JW, Zahedi RP. What can proteomics tell us about platelets? Circ Res. 2014 Mar 28;114(7):1204-19. doi: 10.1161/CIRCRESAHA.114.301598.
2. Loroch S, Kopczynski D, Schneider AC, Schumbrutzki C, Feldmann I, Panagiotidis E, Reinders Y, Sakson R, Solari FA, Vening A, Swieringa F, Heemskerk JWM, Grandoch M, Dandekar T, Sickmann A. Toward Zero Variance in Proteomics Sample Preparation: Positive-Pressure FASP in 96-Well Format (PF96) Enables Highly Reproducible, Time- and Cost-Efficient Analysis of Sample Cohorts. J Proteome Res. 2022 Apr 1;21(4):1181-1188. doi: 10.1021/acs.jproteome.1c00706.