posted on 2013-11-08, 00:00authored byRyan T. Pitman, Jason T. Fong, Penny Billman, Neelu Puri
Recent studies suggest that FTO variants strongly correlate with obesity and mainly influence energy intake with little effect
on the basal metabolic rate. We suggest that FTO influences eating behavior by modulating intracellular energy levels and
downstream signaling mechanisms which control energy intake and metabolism. Since FTO plays a particularly important
role in adipocytes and in hypothalamic neurons, SH-SY5Y neuronal cells and 3T3-L1 adipocytes were used to understand
how siRNA mediated knockdown of FTO expression alters cellular energy homeostasis. Cellular energy status was evaluated
by measuring ATP levels using a luminescence assay and uptake of fluorescent glucose. FTO siRNA in SH-SY5Y cells
mediated mRNA knockdown (282%), increased ATP concentrations by up to 46% (P = 0.013) compared to controls, and
decreased phosphorylation of AMPk and Akt in SH-SY5Y by 252% and 246% respectively as seen by immunoblotting. In
contrast, FTO siRNA in 3T3-L1 cells decreased ATP concentration by 293% (p,0.0005), and increased AMPk and Akt
phosphorylation by 204% and 70%, respectively suggesting that FTO mediates control of energy levels in a cell-type specific
manner. Furthermore, glucose uptake was decreased in both SH-SY5Y (251% p = 0.015) and 3T3-L1 cells (230%,
p = 0.0002). We also show that FTO knockdown decreases NPY mRNA expression in SH-SY5Y cells (221%) through
upregulation of pSTAT3 (118%). These results provide important evidence that FTO-variant linked obesity may be associated
with altered metabolic functions through activation of downstream metabolic mediators including AMPk.
Funding
The authors would like to acknowledge the Research Open Access Publishing (ROAAP) Fund of the University of Illinois at Chicago for financial support
towards the open access publishing fee for this article.