title: Gene-chip studies of adipogenesis-regulated microRNAs in mouse primary adipocytes and human obesity.
abstract: BACKGROUND
  Adipose tissue abundance relies partly on the factors that regulate adipogenesis, i.e. proliferation and differentiation of adipocytes. While components of the transcriptional program that initiates adipogenesis is well-known, the importance of microRNAs in adipogenesis is less well studied. We thus set out to investigate whether miRNAs would be actively modulated during adipogenesis and obesity.
  
  
  METHODS
  Several models exist to study adipogenesis in vitro, of which the cell line 3T3-L1 is the most well known, albeit not the most physiologically appropriate. Thus, as an alternative, we produced EXIQON microarray of brown and white primary murine adipocytes (prior to and following differentiation) to yield global profiles of miRNAs.
  
  
  RESULTS
  We found 65 miRNAs regulated during in vitro adipogenesis in primary adipocytes. We evaluated the similarity of our responses to those found in non-primary cell models, through literature data-mining. When comparing primary adipocyte profiles, with those of cell lines reported in the literature, we found a high degree of difference in 'adipogenesis' regulated miRNAs suggesting that the model systems may not be accurately representing adipogenesis. The expression of 10 adipogenesis-regulated miRNAs were studied using real-time qPCR and then we selected 5 miRNAs, that showed robust expression, were profiled in subcutaneous adipose tissue obtained from 20 humans with a range of body mass indices (BMI, range = 21-48, and all samples have U133+2 Affymetrix profiles provided). Of the miRNAs tested, mir-21 was robustly expressed in human adipose tissue and positively correlated with BMI (R2 = 0.49, p < 0.001).
  
  
  CONCLUSIONS
  In conclusion, we provide a preliminary analysis of miRNAs associated with primary cell in vitro adipogenesis and demonstrate that the inflammation-associated miRNA, mir-21 is up-regulated in subcutaneous adipose tissue in human obesity. Further, we provide a novel transcriptomics database of EXIQON and Affymetrix adipocyte profiles to facilitate data mining.
authors.affiliation: Royal Veterinary College, University of London, UK. jamie.timmons@gmail.com.
authors.forename: Pernille
authors.initials: P
authors.lastname: Keller
authors.forename: Valentina
authors.initials: V
authors.lastname: Gburcik
authors.forename: Natasa
authors.initials: N
authors.lastname: Petrovic
authors.forename: Iain J
authors.initials: IJ
authors.lastname: Gallagher
authors.forename: Jan
authors.initials: J
authors.lastname: Nedergaard
authors.forename: Barbara
authors.initials: B
authors.lastname: Cannon
authors.forename: James A
authors.initials: JA
authors.lastname: Timmons