Long-term exposure of human endothelial cells to metformin modulates miRNAs and isomiRs

Angelica Giuliani,Jacopo Sabbatinelli,Massimiliano Bonafè,Fiorella Marcheselli,Rina Recchioni,Emanuela Mensà,Fabiola Olivieri,Deborah Ramini,Francesco Prattichizzo,Eric Londin,Manuela Ferracin,Isidore Rigoutsos,Maria Rita Rippo

doi:10.1038/s41598-020-78871-5

Abstract

Increasing evidence suggest that the glucose-lowering drug metformin exerts a valuable anti-senescence role. The ability of metformin to affect the biogenesis of selected microRNAs (miRNAs) was recently suggested. MicroRNA isoforms (isomiRs) are distinct variations of miRNA sequences, harboring addition or deletion of one or more nucleotides at the 5′ and/or 3′ ends of the canonical miRNA sequence. We performed a comprehensive analysis of miRNA and isomiR profile in human endothelial cells undergoing replicative senescence in presence of metformin. Metformin treatment was associated with the differential expression of 27 miRNAs (including miR-100-5p, -125b-5p, -654-3p, -217 and -216a-3p/5p). IsomiR analysis revealed that almost 40% of the total miRNA pool was composed by non-canonical sequences. Metformin significantly affects the relative abundance of 133 isomiRs, including the non-canonical forms of the aforementioned miRNAs. Pathway enrichment analysis suggested that pathways associated with proliferation and nutrient sensing are modulated by metformin-regulated miRNAs and that some of the regulated isomiRs (e.g. the 5′ miR-217 isomiR) are endowed with alternative seed sequences and share less than half of the predicted targets with the canonical form. Our results show that metformin reshapes the senescence-associated miRNA/isomiR patterns of endothelial cells, thus expanding our insight into the cell senescence molecular machinery.

Highlights

Increasing evidence suggest that the glucose-lowering drug metformin exerts a valuable antisenescence role
Based on the evidence that treatment with metformin can modulate in vitro cellular senescence, as well as the biogenesis of miRNAs12, we performed for the first time a miR-seq analysis of human umbilical vein endothelial cells (HUVECs) undergoing replicative senescence in the presence of pharmacologically pertinent doses of metformin in order to identify senescence-associated (SA) miRNA and isomiR signatures affected by metformin treatment
To identify the pool of SA miRNAs modulated by metformin, we used a well-established model of human umbilical vein endothelial cells (HUVECs) undergoing replicative s enescence[21,22]

Summary

Introduction

Increasing evidence suggest that the glucose-lowering drug metformin exerts a valuable antisenescence role. While metformin has been shown to influence the cellular epigenetic machinery by modulating Sirtuin-1 (SIRT1)[9], i.e. the pro-longevity histone deacetylase[10], few studies have attempted to identify changes in expression profiles of microRNAs (miRNAs, miRs) induced by metformin treatment in the framework of cellular senescence, mainly showing a general increased abundance of multiple miRNAs after a short-term, high-dose treatment[11,12]. Due to their capability of preventing translation of specific messenger RNAs (mRNA), miRNAs, can impact many cellular processes, including cellular senescence[13]. Based on the evidence that treatment with metformin can modulate in vitro cellular senescence, as well as the biogenesis of miRNAs12, we performed for the first time a miR-seq analysis of human umbilical vein endothelial cells (HUVECs) undergoing replicative senescence in the presence of pharmacologically pertinent doses of metformin in order to identify senescence-associated (SA) miRNA and isomiR signatures affected by metformin treatment

Methods

Results

Conclusion