*Re-Print: The NS-3 Mixture of δ-Tocotrienol, Vitamin D3 and Resveratrol Modulates Gene Expression of Several Novel MicroRNAs Identified by Transcriptomic Analyses in People with Type 2 Diabetes

Aims: Type 2 diabetes mellitus is due to hyperglycemia, therefore fasting glucose and glycosylated hemoglobin (HbA1c) levels are used as biomarkers to determine onset of diabetes. RT-PCR estimation of pooled total mRNAs of EDTA treated whole blood (plasma) obtained after the treatment of NS-3 mixture of d-tocotrienol, vitamin D3, resveratrol of people with type 2 diabetes mellitus (T2DM) for 24 weeks, showed significant down-regulation of gene expression of several diabetes biomarkers (IRS-1, SOD-2, GCKR, IGFBP-2) and cytokines (IL-4, IL-6) as compared to pre-treatment values. Present study investigates the effectiveness of NS-3 on gene expression of mRNAs, miRNAs, and paired mRNA-miRNA in people with T2DM. Methods: Present study is an extension of a randomized placebo controlled double-blinded clinical trial of T2DM (n = 56/group) given two capsules/d of cellulose/olive oil (placebo), or NS-3 for 24-weeks. Pure mRNAs and miRNAs of plasma of pre-dose versus post-dose of NS-3 treated samples were analyzed by next generation sequencing (NGS). Data was uploaded into “Ingenuity Pathways Analyses”. Results: A total of 4000 genes are considered significant, based on > 2-fold gene expression changes. Out of which 1373 genes are significantly differentially expressed in pre-dose vs post-dose (P < 0.02) samples, 20 are up-regulated and 27 are down-regulated of NS-3 treated RNAs of T2DM. Gene expression of up-regulated miR-29b-3p modulates (GLUT4, insulin resistance), miR-624-5p (nephropathy biomarker), miR-361-5p (chronic inflammation), miR-130a-3p (glucose metabolism, insulin secretion), miR-3912-3p (lipid metabolism), and miR-11401 (cellular transcription). The miR-374c-5p (insulin resistance), miR-4326 (HbA1c level)), miR-874-3p (b-cell function) are down-regulated of NS-3 treated people with T2DM. Whereas messengerR-ML-1621513 (oxidative/stress), mR-CTD-2349P217 (insulin-mediated glucose-uptake), are up-regulated, and mR-CTC-246B1810 (b-cell/biology) are down-regulated in T2DM after NS-3 treatment. Venn diagrams have established genetic regulatory network images and canonical signaling pathways for mRNA, miRNA, and paired mRNA-miRNA of gene expression profiles of pre-dose vs post-dose of NS-3 treatment group. Conclusions: The NS-3 treatment of people with T2DM indicates up- or down-regulation of several new miRNAs (miR-29b-3p, miR-624-5p, miR-361-5p, miR-130a-3p, miR-3912-3p, miR-374c-5p, miR-4326 [HbA1c], miR-1247-3p, miR-874-5p) which may be used to identify onset of T2DM. Overexpression of mRNA-AL1621513 indicates oxidative stress in people with T2DM, resulting in complications of diabetes (neuropathy, retinopathy, and stroke).