EPIGENOME-WIDE DNA METHYLATION ANALYSIS OF RAMADAN FASTING EFFECTS ON PEDIATRIC MASLD

Abstract

Metabolic dysfunction associated steatotic liver disease (MASLD) is the most common chronic liver disease in children and is closely linked to insulin resistance (IR). Ramadan fasting (RF), which resembles an intermittent fasting pattern, has shown metabolic benefits in adults. Its effect on children and molecular determinants remains unclear. In this study, we examined the impact of RF on DNA methylation (DNAm) and cardiometabolic markers in children with MASLD. A longitudinal pilot study followed 27 obese children across four visits (T1 before RF, T2–T3 during RF, and T4 one month after RF). Biochemical tests, body composition, diet, sleep, and physical activity habits were collected at each visit and analyzed overall and at each timepoint using R packages. Blood DNAm was measured using the Illumina EPIC v2 array and analyzed using minfi and chAMP. Correlation analysis investigated associations between DNA methylation modifications and phenotypes. Fasting glucose transiently increased during RF (T1 vs T3, p = 0.039) and normalized post-RF (T1 vs T4, p = 0.973). Fasting insulin and HOMA-IR were significantly higher one month after RF (T1–T4, p = 0.033 and p = 0.042, respectively), suggesting a T4 metabolic rebound, though overall time effects did not survive FDR correction. Total sugar intake peaked at T2 before dropping post-fasting below the baseline (T1) level (p=0.001). School-night bedtime was significantly delayed during RF (p < 0.001), and increased daytime sleepiness and bedtime problems persisted post-RF, indicating circadian disruption that persisted beyond the fasting month. Multi-block sPLS-DA showed that the methylation block had the clearest separation between timepoints. DMPs (FDR < 0.05, |Δβ| ≥ 0.1) indicated progressive DNAm changes with RF. T4 showed the strongest pathway changes, with DMPs linked to neuronal signaling, small GTPase cycles, and metabolic pathways including PI3K–Akt, AMPK, and calcium signaling. A core of 51 genes was identified as affected both during and after Ramadan and correlating with diet, circadian disruption, and metabolic outcomes. Overall, RF is associated with progressive DNAm remodeling and a metabolic rebound in children with MASLD, with the post-fasting period showing the most disease-relevant epigenetic changes.

Date of Award	2026
Original language	American English
Awarding Institution	HBKU College of Health & Life Sciences