Identification and validation of key biomarkers of the glycolysis-ketone body metabolism in heart failure based on multi-omics and machine learning.

For qPCR validation, we selected genes representing different functional categories within the WGCNA red module: (1) TIMP1—the hub gene identified by machine learning; (2) HCLS1 and C5AR1—two of the five candidate genes with known immune regulatory functions; (3) THBS4 (Thrombospondin 4)—although not among the final five candidate genes, THBS4 was a highly significant DEG (log2FC = 8.36, p < 0.0001) within the red module with established roles in extracellular matrix remodeling and cardiac fibrosis ( 28 ).

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