Next generation sequencing of RNA reveals novel targets of resveratrol with possible implications for Canavan disease

Abstract

Resveratrol (RSV) is a small compound first identified as an activator of sirtuin 1 (SIRT1), a key factor in mediating the effects of caloric restriction. Since then, RSV received great attention for its widespread beneficial effects on health and in connection to many diseases. RSV improves the metabolism and the mitochondrial function, and more recently it was shown to restore fatty acid βoxidation (FAO) capacities in patient fibroblasts harboring mutations with residual enzyme activity. Many of RSV's beneficial effects are mediated by the transcriptional coactivator PGC-1α, a direct target of SIRT1 and a master regulator of the mitochondrial fatty acid oxidation. Despite numerous studies RSV's mechanism of action is still not completely elucidated. Our aim was to investigate the effects of RSV on gene regulation on a wide scale, possibly to detect novel genes whose upregulation by RSV may be of interest with respect to disease treatment. We performed Next Generation Sequencing of RNA on normal fibroblasts treated with RSV. To investigate whether the effects of RSV are mediated through SIRT1 we expanded the analysis to include SIRT1knockdown fibroblasts. We identified the aspartoacylase (ASPA) gene, mutated in Canavan disease, to be strongly up-regulated by RSV in several cell lines, including Canavan disease fibroblasts. We further link RSV to the up-regulation of other genes involved in myelination including the glial specific transcription factors POU3F1, POU3F2, and myelin basic protein (MBP). We also observe a strong up-regulation by RSV of the riboflavin transporter gene SLC52a1. Mutations in SLC52a1 cause transient multiple acyl-CoA dehydrogenase deficiency (MADD). Our analysis of alternative splicing identified novel metabolically important genes affected by RSV, among which is particularly interesting the α subunit of the stimulatory G protein (Gsα), which regulates the cellular levels of cAMP through adenylyl cyclase. We conclude that in fibroblasts RSV stimulates the PGC-1α and p53 pathways, and up-regulates genes affecting the glucose metabolism, mitochondrial β-oxidation, and mitochondrial biogenesis. We further confirm that RSV might be a relevant treatment in the correction of FAO deficiencies and we suggest that treatment in other metabolic disorders including Canavan disease and MADD might be also beneficial.

Genome-Wide Transcriptional Profiling Reveals PHACTR1 as a Novel Molecular Target of Resveratrol in Endothelial Homeostasis

“…We found that SLC52A1 and CYP1A1 were the most significantly up- and downregulated gene in RES-treated human endothelial cells, respectively. This was consistent with the RNA sequencing results observed in fibroblasts, which showed that SLC52A1 was significantly up-regulated after RES treatment [ 58 ]. Studies have shown that CYP1A1 may be involved in the pathogenesis of atherosclerosis and the occurrence of diabetes mellitus and its vascular complications [ 59 ].…”

Glut 1 in Cancer Cells and the Inhibitory Action of Resveratrol as A Potential Therapeutic Strategy

“…SIRT6 influences the expression of several glycolytic genes such as GLUT1, aldolase, pyruvate dehydrogenase kinase 1 (PDK1), and phosphofructokinase 1 (PFK1) [134]. Furthermore, RSV is known as an activator of SIRT1 and has been related to diabetes progress as a target for its treatment [135,136], inflammation, and neuroprotection [137]. There are data suggesting that RSV has hypoglycemic properties in diabetic rats and restore glycolytic enzyme activities [138,139].…”

Nuclear Peroxisome Proliferator-Activated Receptors (PPARs) as Therapeutic Targets of Resveratrol for Autism Spectrum Disorder

“…Although safety profiles of TZD were documented in young patients with type 1 diabetes [110], further studies are required to understand the effects on behavior and long-term safety of TZD in children with ASD. Novel approaches using natural FAO activators like RSV are particularly appealing, due to their proven application for correction of CPT2- and VLCAD-deficiency in patient cells [103,117].…”