<b>High-frequency feeding with late-night snacking accelerates type 2 diabetes and alters diurnal metabolic gene expression in male KK</b><sup><strong>Ay</strong></sup><b> mice</b>

<jats:p>&lt;p dir="ltr"&gt;We investigated whether sustained high-frequency feeding (six meals per day) increases T2DM incidence and alters diurnal metabolic patterns in male KK&lt;sup&gt;Ay&lt;/sup&gt; mice. Mice were randomly assigned to either three or six isocaloric meals per day for 60 days. Metabolic phenotyping, tissue glycogen assays, multi-tissue circadian clock gene analysis, and exploratory correlation analyses of gene co-expression were performed. Our results demonstrated that high-frequency feeding significantly increased body weight, fasting glucose levels, and T2DM incidence (relative risk = 2.1) compared to three-meal feeding. Six-meal feeding exhibited larger diurnal glucose/insulin oscillations, earlier and higher hepatic glycogen accumulation, disrupted muscle glycogen rhythmicity, and elevated expression of hepatic metabolic genes. Exploratory correlation analyses revealed altered diurnal co-expression patterns among clock and metabolic genes, though causal or regulatory inferences are not warranted from mRNA-level data alone. These findings support meal frequency as an independent modulator of metabolic health and provide preclinical evidence relevant to chrononutrition strategies for T2DM prevention.&lt;/p&gt;</jats:p>