UHPLC and LC-MS/MS Profiling Reveals Gut Microbiota Dysbiosis and Novel Biomarkers in Coronary Heart Disease

While increasing evidence indicates that gut microbiota dysbiosis contributes to coronary heart disease (CHD) pathogenesis through metabolic, inflammatory, and coagulation-related mechanisms, comprehensive multi-omics investigations of individuals with CHD remain limited. In response, a team of researchers aimed to characterize the multi-omics features of CHD and to identify potential diagnostic biomarkers. Non-targeted metabolomic profiling was performed using ultra-high-performance liquid chromatography (UHPLC) coupled with orbital trap mass spectrometry (MS), and quantitative proteomic analysis was conducted using liquid chromatography-tandem mass spectrometry (LC-MS/MS). A paper based on their work was published in Frontiers in Microbiology.¹

Approximately 18.6 million people die from cardiovascular diseases each year, with CHD, a major cause of mortality and disability worldwide, accounting for nearly 45% of these deaths.² While clinical interventions such as statins for lipid regulation, antiplatelet therapy, and revascularization techniques have significantly improved the management of CHD through the slowing of disease progression and enhancement of patient outcomes,^3-5 there are still challenges in the accurate detection of the disease in its early stages, as well as enabling individualized treatment, and reducing the readmission rates. “To further improve CHD prevention and management,” write the authors of the paper,¹ “identifying molecular biomarkers and therapeutic targets that can facilitate early diagnosis and precision interventions through an in-depth understanding of the underlying molecular mechanisms of the disease is essential.”

Recent evidence has spotlighted the crucial role of the gut microbiota and its metabolites in the pathogenesis and progression of CHD.^6,7 Trimethylamine (TMA), which is generated by gut microbiota metabolism of dietary choline and carnitine, is oxidized in the liver to form trimethylamine N-oxide (TMAO, which promotes atherosclerosis progression, enhances platelet reactivity, and increases the risk of adverse cardiovascular events.^8,9 Alterations in the composition and function of the gut microbiota, therefore, may, in the opinion of the research team, provide critical biological information for early detection and stratification of CHD.¹

For this study, blood and fecal samples were collected 10 patients with clinically diagnosed CHD, as well as10 healthy controls, for analysis. The gut microbiota composition was assessed using 16S ribosomal RNA high-throughput sequencing, and shotgun metagenomic sequencing was further performed to evaluate microbial functional potential through the Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation and differential pathway analysis. Functional interaction networks between differentially expressed metabolites and proteins were constructed using Spearman correlation analysis, and the diagnostic potential of candidate biomarkers was evaluated using receiver operating characteristic (ROC) curve analysis.¹

The researchers report that, at the phylum level, the CHD group displayed an increased abundance of the mitrochondria Pseudomonadota, and a decreased abundance of the mitrochondria Bacillota and Actinomycetota. At the genus level, Escherichia-Shigella, Bacteroides, and Klebsiella were significantly enriched; conversely, Bifidobacterium and Faecalibacterium were decreased in abundance. Shotgun metagenomic analysis showed functional remodeling of gut microbiota in CHD, with upregulation of KEGG pathways related to energy metabolism, inflammatory signaling, and host-microbe interactions. Serum metabolomics and proteomic analyses revealed 32 differentially expressed metabolites and 38 differentially expressed proteins, respectively. Correlation analysis showed significant associations between phospholipid metabolites and apolipoproteins, inflammatory mediators and the complement system, asymmetric dimethylarginine and endothelial function-related proteins, and oxidative stress metabolites and antioxidant proteins. ROC analysis identified several potential biomarkers with high diagnostic value.¹

The authors of the paper state that the study demonstrates “that individuals with CHD exhibit significant gut microbiota dysbiosis, distinct metabolic pathway alterations, and aberrant expression of coagulation- and inflammatory-related proteins. These findings provide novel insights into potential targets for CHD prevention and treatment strategies.”¹

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References

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2. WHO Cardiovascular Diseases Fact Sheets [Online]. World Health Organization website 2023. https://www.who.int/health-topics/cardiovascular-diseases (cccessed 2025-10-28).
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7. Escobar, C.; Aldeguer, X.; Vivas, D. et al. The Gut Microbiota and its Role in the Development of Cardiovascular Disease. Expert Rev Cardiovasc Ther. 2025, 23 (1-2), 23-34. DOI: 10.1080/14779072.2025.2463366
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9. Koeth, R. A.; Wang, Z.; Levison, B. S. et al. Intestinal Microbiota Metabolism of L-carnitine, a Nutrient in Red Meat, Promotes Atherosclerosis. Nat Med. 2013, 19 (5), 576-585. DOI: 10.1038/nm.3145