Exploring proteoform biomarkers to explain variability in pre-clinical cardiovascular disease risk among male and female young adults with high exposure to adverse childhood experiences

Background

Around 15-20% of the population grow up in a toxic home environment exposed to high levels of adverse childhood experiences (ACEs), such as severe household dysfunction and maltreatment. Research indicates that high exposure to ACEs trigger physiological responses, leading to an array of later conditions including cardiovascular disease (CVD). The pathways linking ACEs to CVD likely involve lifestyle, psychosocial, and inflammatory factors that alter health trajectories through complex molecular mechanisms. While the conceptual linkage between biological, lifestyle, and psychosocial factors as determinants of health is recognized, understanding the precise mechanisms remains challenging. One potential avenue to elucidate these mechanisms is to examine overall differences in biologically active proteins (proteoforms) that link ACEs with pre-clinical markers of CVD, specifically central arterial stiffness (carotid-femoral pulse wave velocity; cfPWV). Comparing young adults with higher vs. lower cfPWV among both males and females with high ACE exposure can identify specific proteoforms that may contribute to variation in pre-clinical CVD risk markers.

Methods

Using 2-dimensional gel electrophoresis (2DE) coupled with liquid chromatography and tandem mass spectrometry (LC/MS/MS), we analyzed the total serum proteoform content separately for male and female participants aged 19-26 with high ACE levels (≥4), stratified across the highest and lowest quartile of cfPWV. Targeted ELISA-based detection platforms will measure the abundance of identified proteoforms in participants with low ACE levels (0) and high ACE levels (≥4). These measurements will then be compared against previously collected data on lifestyle, psychosocial, and inflammatory factors to determine correlations.

Results

Initial findings reveal over 100 resolved serum proteoforms differing significantly between high and low cfPWV groups. Moreover, a significant interaction between cfPWV and sex was identified for some proteoforms indicating potential sex-specific effects. All identifications belonged to various classes, predominantly defense/immunity, protein-binding activity modulators, metabolite interconversion enzymes, and protein modifying enzymes. Notably, some were found in more than one fixed MW and pI gel region, suggesting the presence of different proteoforms.

Conclusion

These proteoform identifications offer insights into pathways and molecular mechanisms linking ACEs to CVD risk. Determining to what extent the concentrations of these markers are influenced by lifestyle, psychosocial, and inflammatory factors may enable the identification of protective and/or risk factors. This will facilitate the development of targeted protein detection tools to validate and evaluate protein-specific biomarkers in ACE-driven development of CVD, contributing to a deeper understanding of this complex relationship.