Integrated metabolomics and machine learning identify potential biomarkers and metabolism-related targets for MASLD in older adults.

BACKGROUND: Given the high prevalence and serious health threats of Metabolic dysfunction-associated steatotic liver disease (MASLD) in older people, uncovering the pathological features of MASLD in the context of age is critically important. This study aimed to perform serum metabolomics analysis for older adults with MASLD to identify potential biomarkers and involved metabolic disturbances. METHODS: 30 older subjects with MASLD and 30 healthy individuals were included (age > 65). Sex-stratified metabolomics analysis was applied by using ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS/MS). Partial least squares discriminant analysis (PLS-DA) was utilized to identify differential metabolites between groups. The distinct and shared pathways of MASLD in older men and older women were explored using MetaboAnalyst 6.0. Machine learning algorithms are applied to screen for optimal biomarkers. Receiver operating characteristics (ROC) curve analysis was applied to test the predictive ability of biomarkers. Spearman's correlation analysis was conducted to evaluate the correlation of metabolic biomarkers with biochemical parameters. The differences in phosphatidylcholine concentration were externally validated in other 60 older adults. Finally, in the aged mice MASLD model, the mRNA expression levels of the rate-limiting enzyme of phosphatidylcholine synthesis (PCYT1A and PEMT) were detected. RESULTS: Principal components analysis (PCA) and PLS-DA exhibited pronounced metabolic separations between the MASLD and HC groups in both older males and females. 14 shared metabolites were selected by the least absolute shrinkage and selection operator (LASSO) and Support Vector Machine-Recursive Feature Elimination (SVM-RFE) algorithms. Finally, 6 metabolites significantly dysregulated in older MASLD patients (|logFC| > 0.5) would be determined as the optimal metabolic markers, including: PC 18:1_22:6, 2'-Deoxyuridine-5-monophosphate, porphobilinogen, uridine, all-trans-13,14-Dihydroretinol and D-Ribulose 1,5-bisphosphate. Notably, a marked difference in serum phosphatidylcholine levels in the elderly between the MASLD group and the healthy group was confirmed by external validation (p < 0.001). The significant downregulation of PCYT1A mRNA expression was also found in the aged mice MASLD model. CONCLUSIONS: This study depicts the metabolic features of MASLD in the older population. Our finding explores promising biomarkers for diagnosis and provides more perspectives on molecular mechanisms and potential targets for MASLD in the context of aging.