Bioinformatic identification in grape seeds RNA-seq data of miRNAs with crosskingdom activity involved in pathological condition associated with aging

Introduction Recent publications show that through our diet, we ingest plant miRNAs which, once absorbed, can travel in the bloodstream, either as free molecules or encapsulated in microvesicles, reaching target tissues and organs where they exert gene regulation. This phenomenon is known as Cross-Kingdom Regulation 1 . Crosskingdom regulation may contribute to various health effects, including potential nutritional benefits or even therapeutic effects. The activity focused on developing a bioinformatics pipeline for a bioinformatic study aimed at identifying miRNAs with cross-kingdom activity in grape seeds. Method Cross-kingdom gene regulation assumes that plant miRNA is accepted from human RISC complex. We have developed Croseed, a bioinformatics pipeline that performs a comparative human/grapevine study, considering both sequence similarity and the structural constraints imposed by the human RISC complex proteins 2 . The study enables the identification of plant miRNAs mimicking the biological function of human endogenous miRNAs, competing for the same target sites on the human RNA messengers. We used Croseed on RNA expression data from grape seeds (PmREN2.0 3 ) to identify miRNAs with cross-kingdom activity Results 45 bioactive miRNAs expressed in the seeds have been identified and proximately 395 human genes could potentially undergo Cross-Kingdom regulation. The gene network analysis using the databases Kegg 4 , miRTarbase 5 and PmREN2.0 pointed out 26 genes involved in neurological disease (Alzheimer Disease and senescence). Moreover, hub genes (MTOR, PPARA, PTEN, STAT3) associated with obesity and insulin resistance have been shown to be down regulated by several miRNAs expressed in grape seed. The results indicate that grape seeds could be a source of bioactive miRNAs to be used in functional foods. Perspective 3T3-L1 cell line are fibroblasts, isolated from mouse embryo, that can be used to study the basic cellular mechanisms associated with diabetes, obesity and related disorders. We are testing the effects of the grape seed miRNAs on 3T3-L1 cells and assessing the effects through differential gene expression studies.