Low-density lipoprotein receptor genotypes modify the sera metabolome of patients with homozygous familial hypercholesterolemia

Figures

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  Figure 1. Study design and multivariate statistical pattern analysis of metabolomic data (A) Grouping design of the study patients. (B) Treemap overview for the category distribution of identified metabolites. (C and D) PCA and PLS-DA scores plot of the metabolomic data from serum samples of patients with TH and CH; each point represents an individual serum sample. (E) PLS-DA permutations plot of metabolomic data of TH and CH groups; each green point and black point represents R2-value and Q2-value, respectively. The criteria for PLS-DA model validity: All Q2-values to the left are lower than the original points to the right; The left regression line of the Q2-points is below zero; all R2-values to the left are lower than the original point to the right. (F–H) PCA scores plot, PLS-DA scores plot, PLS-DA permutations plot of the metabolomic data from defective/defective, null/defective, and null/null groups.

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  Figure 2. Metabolomic signatures associated with LDLR functional status (A) VIP plots of each metabolite variation in pairwise comparisons of defective/defective, null/defective, and null/null groups; the dashed lines indicate the threshold for the significant differences. (B) Hierarchical clustering heatmap of LDLR status-associated metabolite intensities in different groups. (C) PCA scores plot of the differentiated metabolites; each point represents an individual sample. CE = cholesterol esters, TAG = triacylglycerides, LysoPC = lysophosphocholines, LysoPC O = alkyl LysoPC, LysoPE O = alkyl lysophosphoethanolamines, LysoPA = lysophosphatidic acid, LPI = lysophosphatidylinositol.

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  Figure 3. Metabolic pathway enrichment and regression analyses (A) Pathway enrichment analysis of the differentiated metabolites that distinguished the study groups; the bubble size refers to the enrichment factor of the pathway and the color represents the natural log of the pathway p value. (B) Correlation plot depicting the relation of each metabolite and LDLR status; positive or negative β-coefficient values indicate a positive or negative correlation between metabolites and LDLR status (at least LDLR null mutation: null/null or null/defective), FDR-adjusted p < 0.05 was considered as significant threshold, n.s. represents no significance; Model 1: Adjustments for ages, gender, and CVD history; Model 2: Adjustments for all variables in model 1 and clinical lipids (TC, TG, LDL-C, HDL-C, Lp(a), and APOB). NN = null/null, DD = defective/defective, ND = null/defective. Other abbreviations are seen in Figure 2.

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  Figure 4. Differential metabolites correlated with clinical lipids (A) Spearman’s rank correlation heatmap depicting the relationship of each LDLR status-associated metabolites with clinical lipids. Positive correlations are displayed in red and negative correlations in blue. Color intensity and size of the circle are proportional to the Spearman correlation coefficients. In this correlogram, correlations with p < 0.05∗ or p < 0.01∗∗ are considered significant. (B) Debiased sparse partial correlation network plot visualizing the major relationships of clinical lipids and key metabolites; circles represent metabolites, squares represent clinical lipids, the solid lines indicate positive correlation, the dashed lines indicate negative correlation, the thickness of the edges represents the strength of the partial correlation coefficients (|coefficients| ranges from 0.5 to 1.0). Abbreviations are seen in Figure 2.