MicroRNA profiling reveals age-dependent differential expression of nuclear factor κB and mitogen-activated protein kinase in adipose and bone marrow-derived human mesenchymal stem cells

Abstract

IntroductionMesenchymal stem cells (MSCs) play a central role in mediating endogenous repair of cell and tissue damage. Biologic aging is a universal process that results in changes at the cellular and molecular levels. In the present study, the role of microRNA (miRNA) in age-induced molecular changes in MSCs derived from adipose tissue (ASCs) and bone marrow (BMSCs) from young and old human donors were investigated by using an unbiased genome-wide approach.MethodsHuman ASCs and BMSCs from young and old donors were cultured, and total RNA was isolated. The miRNA fraction was enriched and used to determine the expression profile of miRNA in young and old donor MSCs. Based on miRNA expression, differences in donor MSCs were further investigated by using differentiation assays, Western blot, immunocytochemistry, and bioinformatics.ResultsBiologic aging demonstrated reduced osteogenic and adipogenic potential in ASCs isolated from older donors, whereas cell size, complexity, and cell-surface markers remained intact with aging. Analysis of miRNA profiles revealed that small subsets of active miRNAs changed secondary to aging. Evaluation of miRNA showed significantly decreased levels of gene expression of inhibitory kappa B kinase (IκB), interleukin-1α, inducible nitric oxide synthase (iNOS), mitogen-activated protein kinase/p38, ERK1/2, c-fos, and c-jun in MSCs from older donors by both bioinformatics and Western blot analysis. Nuclear factor kappa B (NF-κB), myc, and interleukin-4 receptor mRNA levels were significantly elevated in aged cells from both the adipose and bone marrow depots. Immunocytochemistry showed nuclear localization in young donors, but a cytosolic predominance of phosphorylated NF-κB in ASCs from older donors. Western blot demonstrated significantly elevated levels of NF-κB subunits, p65 and p50, and AKT.ConclusionsThese findings suggest that differential expression of miRNA is an integral component of biologic aging in MSCs.

Adipose-Derived Mesenchymal Stromal Cells From Aged Patients With Coronary Artery Disease Keep Mesenchymal Stromal Cell Properties but Exhibit Characteristics of Aging and Have Impaired Angiogenic Potential

“…There were no discernible age-associated changes in MSC marker profiles and ADSC differentiation potential, which correlated with results of Pandey et al [43].…”

Effects of Aging on the Collateral Circulation, and Therapeutic Implications

“…Another potential therapeutic-related approach might involve targeting the increased oxidative stress occurring with aging. Thus, Miller and coworkers 46 examined the capacity of the ileal artery of aged rats to remodel, in vivo, in response to ligation of adjacent arteries. They found that luminal expansion was absent in aged rats, but that expansion was restored by antioxidant treatment.…”

Influence of Donor Age and Passage Number on Angiogenic Activity in Human Adipose-Derived Stem Cell-Conditioned Media

“…A similar donor age-dependent decline in VEGF levels has been reported in mouse and rat bone marrow MSCs [18,43] and mouse ASCs [26]. A recent study using human bone marrow MSCs and ASCs demonstrated donor age-associated changes in microRNAs, which regulate expression of several genes and cellular function [23,24]. Further studies are needed to identify the essential molecules, including micro-RNAs, that regulate biological age-dependent VEGF-A expression.…”

“…The proliferation and differentiation potential of MSCs/ASCs is known to be affected by in vitro aging (passage number in culture) [18][19][20][21] and by in vivo aging (donor age) [18,20,[22][23][24][25]. On the other hand, limited information is currently available regarding the angiogenic potential of aged ASCs [26][27][28].…”