TARGETED METABOLOMIC PROFILING REVEALS DISTINCT PATHWAY ALTERATIONS IN FAMILIAL HYPERCHOLESTEROLEMIA

Therapeutic Area

Novel Biomarkers

Background

Familial hypercholesterolemia (FH) is a monogenic lipid disorder characterized by markedly elevated low-density lipoprotein cholesterol and a high risk of premature atherosclerosis. However, LDLC alone does not fully account for the metabolic complexity and residual cardiovascular risk observed in FH. In this study, we performed targeted metabolomic profiling to uncover systemic metabolic alterations and identify pathways that may contribute to disease pathogenesis and cardiovascular outcomes.

Methods

Plasma samples from 16 FH patients and 15 non-FH controls were analyzed using a SCIEX Triple Quad™ 7500 LC-MS/MS-QTRAP with an ExionLC system. Chromatographic separation was performed on a Kinetex F5 column using a water/acetonitrile gradient with 0.1% formic acid. A Scheduled MRM™ method in positive and negative ion modes targeted over 800 metabolites. Instrument settings included a 350°C ion source temperature and optimized gas flows. Peaks were quantified using Analyst 3.1 software and normalized to internal standards. Data processing and pathway analysis were conducted using MetaboAnalyst 6.0 and GraphPad Prism 10.

Results

Targeted metabolomic profiling detected 248 plasma metabolites, of which 72 were significantly downregulated and 9 upregulated in FH patients compared to healthy controls. PCA and PLS-DA demonstrated clear group separation, and volcano plot and heatmap analyses confirmed distinct differences in metabolite abundance. Pathway enrichment analysis revealed 16 significantly disrupted metabolic pathways in FH (Figure 1), including primary bile acid biosynthesis (p = 0.0005), beta-alanine metabolism (p = 0.0013), arginine and proline metabolism (p = 0.0013), glutathione metabolism (p = 0.0087), and pyrimidine metabolism (p = 0.0097). Several of the altered pathways were linked to amino acid metabolism, oxidative stress, and nucleotide synthesis, indicating broad systemic metabolic disturbances in FH beyond lipid dysregulation.

Conclusions

Our targeted metabolomic analysis revealed broad metabolic dysregulation in FH patients, including significant alterations in bile acid biosynthesis, amino acid metabolism, nucleotide pathways, and oxidative stress processes. This work lays the groundwork for future studies to identify novel biomarkers and therapeutic targets for cardiovascular risk reduction in FH.