Abstract

BackgroundMesenchymal stem cells including adipose-derived stem cells (ASCs) have a considerable potential in the field of translational medicine. Unfortunately, multiple factors (e.g., older age, co-existing diabetes, and obesity) may impair cellular function, which hinders the overall effectiveness of autologous stem cell therapy. Noncoding RNAs—including microRNAs (miRNAs), long ncRNAs (lncRNAs), and circular RNAs (circRNAs)—have been shown to play important roles in stem cell biology. However, the overall diabetes-related and aging-related expression patterns and interactions of these RNAs in ASCs remain unknown.MethodThe phenotypes and functions of ASCs isolated from diabetic (D-ASCs), old (O-ASCs), and young (Y-ASCs) donors were evaluated by in vitro assays. We conducted high-throughput RNA sequencing (RNA-seq) in these ASCs to identify the differentially expressed (DE) RNAs. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and protein-protein interaction (PPI) analyses were performed to investigate mRNAs with significant differences among groups. The lncRNA- or circRNA-associated competing endogenous RNA (ceRNA) networks were constructed based on bioinformatics analyses and real-time polymerase chain reaction (RT-PCR) results. The miR-145-5p mimics were transfected into O-ASCs and verified by PCR.ResultsASCs from diabetic and old donors showed inferior migration ability and increased cellular senescence. Furthermore, O-ASCs have decreased capacities for promoting endothelial cell angiogenesis and fibroblast migration, compared with Y-ASCs. The DE miRNAs, mRNAs, lncRNAs, and circRNAs were successfully identified by RNA-seq in O-ASCs vs. Y-ASCs and D-ASCs vs. O-ASCs. GO and KEGG analyses demonstrated that DE mRNAs were significantly enriched in aging and cell senescence terms separately. PPI networks revealed critical DE mRNAs in the above groups. RNAs with high fold changes and low p values were validated by PCR. ceRNA networks were constructed based on bioinformatics analyses and validated RNAs. Additionally, the lncRNA RAET1E-AS1–miR-145-5p–WNT11/BMPER axis was validated by PCR and correlation analyses. Finally, the overexpression of miR-145-5p was found to rejuvenate O-ASCs phenotype and augment the functionality of these cells.ConclusionOur research may provide insights regarding the underlying mechanisms of ASC dysfunction; it may also offer novel targets for restoring therapeutic properties in ASCs.

Highlights

  • Mesenchymal stem cells including adipose-derived stem cells (ASCs) have a considerable potential in the field of translational medicine

  • We demonstrated that the long ncRNAs (lncRNAs) RAET1E-AS1– miR-145-5p–WNT11/BMP binding endothelial regulator (BMPER) axis may play a vital role in regulating ASC senescence and function

  • We found that four miRNAs, 22 mRNAs, eight Circular RNA (circRNA) were upregulated in O-ASCs, compared with DASCs; these RNAs were all upregulated in Y-ASCs, Fig. 1 Evaluation of the phenotype and function of ASCs isolated from diabetic, old, and young patients. a EdU assay analysis of the proliferation rate of ASCs from those three groups

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Summary

Introduction

Mesenchymal stem cells including adipose-derived stem cells (ASCs) have a considerable potential in the field of translational medicine. The most recent clinical successes involving limbal stem cells in corneal restoration and transgenic stem cells for epidermis regeneration have been compelling [1, 2], which presumably reflect the considerable potential of this field. Most of these studies remain in preclinical or clinical trial stages. The therapeutic effect of stem cells can be impaired by physical conditions within donors (e.g., high age, diabetes, obesity, and chronic kidney disease) [7,8,9]. Given the high morbidity of diabetes and the rapidly aging global population, the donor condition merits extensive consideration

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