title: 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: BACKGROUND
  Mesenchymal 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.
  
  
  METHODS
  Human 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.
  
  
  RESULTS
  Biologic 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.
  
  
  CONCLUSIONS
  These findings suggest that differential expression of miRNA is an integral component of biologic aging in MSCs.
authors.affiliation: Center for Stem Cell Research and Regenerative Medicine, School of Medicine, Tulane University, New Orleans, LA, USA.
authors.forename: Amitabh C
authors.initials: AC
authors.lastname: Pandey
authors.forename: Julie A
authors.initials: JA
authors.lastname: Semon
authors.forename: Deepak
authors.initials: D
authors.lastname: Kaushal
authors.forename: Regina P
authors.initials: RP
authors.lastname: O'Sullivan
authors.forename: Julie
authors.initials: J
authors.lastname: Glowacki
authors.forename: Jeffery M
authors.initials: JM
authors.lastname: Gimble
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Bunnell
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000375:Aging; D001854:Bone Marrow Cells / Q000166:cytology; D019295:Computational Biology; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D035683:MicroRNAs / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D014019:Tissue Donors
chemical_list: D016207:Cytokines; D035683:MicroRNAs; D016328:NF-kappa B; D052247:Nitric Oxide Synthase Type II; D020928:Mitogen-Activated Protein Kinases