Supplementary Material for: Dairy Product Intake Modifies MicroRNA Expression among Individuals with Hyperinsulinemia: A Post-Intervention Cross-Sectional Study

<b><i>Introduction:</i></b> MicroRNA (miRNA) profiles have been shown to change after intake of dairy products. Dysregulation of miRNA is associated with the changes in the level of glycemic parameters. The objectives are: (1) to investigate miRNA expression after consumption of dairy products and (2) to study the association between miRNAs and glycemic profile among individuals with hyperinsulinemia. <b><i>Methods:</i></b> In crossover design, 24 participants were randomized into 2 phases: high dairy (HD) (≥4 servings/day according to the Canadian food guide [2007]) and adequate dairy (AD) (≤2 servings/day) over 6 weeks. First, miRNAs were extracted from a pooled plasma sample of 10 subjects after HD and AD intervention which analyzed in duplicate by array hybridization (Affymetrix Gene Chip miRNA Array v. 4.0). Second, 6 miRNAs related to type 2 diabetes (T2D) were validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) from plasma of 24 participants. <b><i>Results:</i></b> Microarray analysis indicated that 297 miRNAs expressed differentially (FC ≥ ±1.2; <i>p</i> value &lt;0.05) in a pooled plasma sample of 10 subjects. Among pooled miRNAs, the level of selected miRNAs, including miR-652-3p, miR-106b-5p, miR-93-5p, and miR-107 were downregulated; conversely, miR-223-3p and miR-122-5p were upregulated. After qRT-PCR validation, only the expression level of miR-106-5p tended to be increased after HD compared to AD (<i>p</i> = 0.06). After AD intervention, the level of fasting plasma glucose (FPG) and insulin and homeostatic model assessment of insulin resistance were negatively correlated with miR-122-5p. Similarly, negative correlation was found between miR-106-5p and FPG. <b><i>Conclusion:</i></b> The miRNAs profile was modified after HD compared to AD, and this may have role in modifying the risk of T2D (registration No. NCT02961179).