Abstract 108: Altered Erk / Klf4-mediated Control Of Agrp And Metabolic Rate During Obesity

Abstract

Resting metabolic rate (RMR) adaptation contributes to the maintenance of human obesity, yet mechanisms controlling RMR in lean vs obese states remain poorly defined. Leptin contributes to RMR control in part through suppression of inhibitory Agouti-related peptide (AgRP) neurotransmission in the hypothalamic arcuate nucleus (ARC). Within AgRP neurons, leptin activates the extracellular signal-regulated kinase (ERK) cascade, which is thought to interrupt Agrp transcription through interference with the Krüppel-like factor 4 (KLF4) transcription factor, and our published data support altered ARC ERK signaling during obesity. We therefore hypothesized a role for ERK/KLF4 signaling within AgRP neurons for the suppression of AgRP transcription and thereby stimulation of RMR in the lean state, and changes in this pathway during obesity. In the GT1-7 cell model of AgRP neurons, leptin increased the phosphorylated ERK (pERK)/ERK ratio (n=4, 1 uM, 15 min +89% p<0.05) and reduced Agrp mRNA (n=4, 1 uM, 1 hr -43.0% p<0.05); similarly, siRNA knockdown of Klf4 reduced Agrp mRNA (n=4, -21.6% p<0.05). In lean adult C57BL/6J mice, leptin increased the ARC pERK/ERK ratio (n=8, 1 ug/g ip 16.5-fold p<0.05) and stimulated RMR (n=11, 2 ug/g ip 30 min +43.8% p<0.05). Lean transgenic mice with conditional deletion of KLF4 from AgRP neurons ( Agrp -Cre x Klf4 flox ) exhibited increased RMR (n=13 KO +13.48% vs controls n=22 p<0.05). These findings support the model that ERK inhibition of KLF4 within AgRP neurons mechanistically contributes to leptin-mediated control of Agrp and RMR in the lean state. Ten weeks of high fat feeding strongly increases endogenous plasma leptin in C57BL/6J mice, and this treatment increased the ARC pERK/ERK ratio (n=5 +54.4% p<0.05), yet ARC KLF4 was increased (n=6 +74.6% p<0.05) and Agrp mRNA was not suppressed (n=4 +1.6-fold, p=ns). Therefore, these data also support the concept that loss of ERK-mediated suppression of KLF4 within the AgRP neuron during obesity may contribute to the pathogenesis of RMR adaptation.