Metabolic syndrome increases senescence-associated micro-RNAs in extracellular vesicles derived from swine and human mesenchymal stem/stromal cells

Abstract

Background The metabolic syndrome (MetS) is a combination of cardiovascular risk-factors, including obesity, hypertension, hyperglycemia, and insulin resistance. MetS may induce senescence in mesenchymal stem/stromal cells (MSC) and impact their micro-RNA (miRNA) content. We hypothesized that MetS also alters senescence-associated (SA) miRNAs in MSC-derived extracellular vesicles (EVs), and interferes with their function. Methods EVs were collected from abdominal adipose tissue-derived MSCs from pigs with diet-induced MetS or Lean controls (n = 6 each), and from patients with MetS (n = 4) or age-matched Lean controls (n = 5). MiRNA sequencing was performed to identify dysregulated miRNAs in these EVs, and gene ontology to analyze their SA-genes targeted by dysregulated miRNAs. To test for EV function, MetS and Lean pig-EVs were co-incubated with renal tubular cells in-vitro or injected into pigs with renovascular disease (RVD, n = 6 each) in-vivo. SA-b-Galactosidase and trichrome staining evaluated cellular senescence and fibrosis, respectively. Results Both humans and pigs with MetS showed obesity, hypertension, and hyperglycemia/insulin resistance. In MetS pigs, several upregulated and downregulated miRNAs targeted 5768 genes in MSC-EVs, 68 of which were SA. In MetS patients, downregulated and upregulated miRNAs targeted 131 SA-genes, 57 of which overlapped with pig-EVs miRNA targets. In-vitro, MetS-MSC-EVs induced greater senescence in renal tubular cells than Lean-MSC-EVs. In-vivo, Lean-MSC-EVs attenuated renal senescence, fibrosis, and dysfunction more effectively than MetS-MSC-EVs. Conclusions MetS upregulates SA-miRNAs in swine MSC-EVs, which is conserved in human subjects, and attenuates their ability to blunt cellular senescence and repair injured target organs. These alterations need to be considered when designing therapeutic regenerative approaches. Graphical abstract

Metabolic Syndrome Alters the Cargo of Mitochondria-Related microRNAs in Swine Mesenchymal Stem Cell-Derived Extracellular Vesicles, Impairing Their Capacity to Repair the Stenotic Kidney

“…Therefore, MetS-induced changes in the miRNA profile of EVs might have exerted important posttranscriptional changes in mitochondria-related genes in recipient cells, which might have contributed to mitochondrial injury and in turn, impaired the capacity of MetS-EVs to repair the stenotic kidney. Importantly, our findings are in agreement with our recent observation that MetS alters the miRNA cargo of human MSC-derived EVs [65], suggesting that MetSinduced changes in the content of EVs are conserved in human subjects. Therefore, miRNAs packed in MSCderived EVs may have important implications for the use of autologous EVs as a therapeutic regenerative strategy.…”

The Micro-RNA Cargo of Extracellular Vesicles Released by Human Adipose Tissue-Derived Mesenchymal Stem Cells Is Modified by Obesity

“…The EV miRNA cargo in all samples was assessed by miRNA sequencing, as described ( Li et al, 2020 ; see Supplementary Material ), after passing quality control tests ( Supplementary Tables 1 , 2 ). Total RNA was extracted from EVs using the exoRNeasy Maxi Kit (cat#77044, Qiagen, Germany).…”

Mesenchymal stem cells and extracellular vesicles in therapy against kidney diseases

“…At the same time, some researchers proposed that MSC-Exos can only partially relieve ageing kidney induced by renal artery stenosis [98]. MetS is able to change the amount of loading of miRNA on EVs, upregulate ageing-related miRNA in EVs, and even limit the use of EVs in exogenous regenerative therapy through abnormal transcription [99][100][101]. Zhao et al found that autologous ADMSCs can better preserve microcirculation through comparative studies, while ADMSC-EVs perform better in retaining intactness of nephrocytes and reducing necrosis [102].…”