Endoplasmic reticulum stress in a circumventricular organ-hypothalamic neuronal circuit alters hepatic glucose and lipid metabolism during obesity
Authors: Kim Han Rae , Tabiatnejad Parisa , Young Colin N. 
Published: May 2024

Non-alcoholic fatty liver disease (NAFLD) is a leading cause of chronic hepatic disorders and is associated with metabolic conditions such type 2 diabetes. Accumulating evidence points to endoplasmic reticulum (ER) stress and activation of the unfolded protein response (UPR) in the subfornical organ (SFO), a circumventricular nucleus situated outside of the blood-brain-barrier, as a major contributor to NAFLD development during obesity. We recently reported that ER stress in SFO neurons that project to the hypothalamic paraventricular nucleus (PVN, SFO→PVN neurons) drives liver triglyceride accumulation during obesity. However, the mechanism by which ER stress in this neural network mediates obesity-related hepatic steatosis remains unknown. Elevations in hepatic gluconeogenesis, lipogenesis, and free fatty acid uptake are all associated with the development of fatty liver. Based on this, we hypothesized that ER stress in SFO→PVN neurons may contribute to NAFLD through alterations in hepatic glucose/lipid metabolism during obesity. To investigate this, we used an intersectional viral strategy to selectively inhibit ER stress in SFO→PVN neurons in C57Bl/6J male mice that were fed a high-fat diet (HFD, 60% kCal fat) for 7 wks (n=4-5/group) and then underwent selective PVN targeting of a retrogradely transported Cre-recombinase virus (CAV2-Cre-GFP; CAV2-GFP served as a control). Concurrently, a Cre inducible vector to overexpress the ER chaperone glucose-regulated peptide 78 (GRP78; AAV-FLEX-GRP78-mStrawberry) was targeted to the SFO. This approach results in the overexpression of GRP78 specifically in SFO→PVN neurons, which has been established as an effective means of reducing ER stress. Four weeks following selective inhibition of SFO→PVN neuronal ER stress, body weight (46±2 vs 43±1 g, CAV2-GFP vs CAV2-Cre, P=0.26), food intake, and adipose mass were not different between groups. However, diminishing UPR activation in this circumventricular-hypothalamic circuit resulted in a reduction in hepatomegaly (1.8±0.1 vs 1.5±0.1 g, CAV2-GFP vs CAV2-Cre, P=0.08), hepatic triglyceride levels (4.3±0.6 vs 2.3±0.4 mM, CAV2-GFP vs CAV2-Cre, p<0.05), and plasma glucose (198±12 vs 168±8 mg/dL, CAV2-GFP vs CAV2-Cre, P=0.06). Interestingly, reducing ER stress in SFO→PVN neurons was also associated with a downregulation in hepatic mRNA levels of genes involved in hepatic gluconeogenesis (e.g. G6pase: 0.29±0.04; Pepck: 0.46±0.04 fold CAV2-GFP, both p<0.05), de novo lipogenesis (e.g. SREBP1: 0.65±0.1 fold CAV2-GFP, P=0.05) and free fatty acid synthesis and uptake (e.g. Fasn: 0.57±0.11 fold CAV2-GFP, P=0.09 and FATP5: 0.42±0.03, fold CAV2-GFP, p<0.05). Overactivation of the liver sympathetic nerves has recently been shown to contribute to obesity-related NAFLD, and inhibition of ER stress in SFO→PVN neurons resulted in a reduction in protein levels of hepatic tyrosine hydroxylase (TH), the rate limiting enzyme in catecholamine synthesis (0.58±0.03, fold CAV2-GFP, p<0.05). Notably, hepatic TH levels were significantly correlated with liver glucose/lipid metabolism-related gene expression and triglyceride content (e.g. Pepck: R2=0.57; Fasn: R2=0.80; FATP5: R2=0.77; triglycerides: R2=0.70, all p<0.05). Collectively, these results suggest that during obesity, ER stress in SFO→PVN neurons may drive liver sympathetic overactivity and subsequently disrupt hepatic glucose/lipid metabolism, thus contributing to NAFLD development. R01DK117007, R01HL141393, AHA932522. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.