Obesity is a public health epidemic wordwide and affects nearly a third of adult Americans. Several devastating co-morbidities are associated with obesity, including insulin resistance/type II diabetes and non-alcoholic steatohepatitis/non-alcoholic fatty liver disease. Paradoxically, in obese states, lipid storage is not suppressed, in spite of resistance to insulin action. This finding that has important consequences for the management of obesity and its complications. An emerging molecular mechanism linking obesity to excessive lipid storage is the mTORC1/SREBP1c pathway, which our group and others have identified as both activated with obesity, and as a key regulator of lipogenesis and new adipocyte formation. This study will test the hypothesis that mTORC1 activation in the obese state elevates lipid levels, due to activation of both adipogenesis and lipogenesis. To accomplish this, we have developed several new innovative models to test specific aspects of this hypothesis. First, we have generated adipose-specificTsc1 knockout mice as a model of chronic adipose mTORC1 activation. The chronic elevations in mTORC1 signaling in these mice are associated with elevated fat mass and increased hepatic steatosis, likely due to enhanced de novo lipogenesis in adipose tissue. We will determine the molecular changes resulting from chronic mTORC1 elevation, and identify the molecular mechanisms underlying these mTORC1-dependent increases in lipid storage. Elevated adiposity may is caused by increased adipogenesis, so we will determine the molecular mechanisms by which mTORC1 positively regulates adipogenesis. We will specifically evaluate the hypothesis that mTORC1 regulates PPARγ mRNA stability via a miRNA-dependent mechanism. To test the role of mTORC1 in the liver, we will study both activation and inhibition this kinase via ablation of the essential mTORC1 component Rptor, and Tsc1 respectively in adult mouse livers. This approach will allow us to evaluate whether mTORC1 is necessary and sufficient for the development and maintenance of hepatic steatosis in adult liver tissues for the first time. This is an important gap in our knowledge, since in obesity-associated liver disease, mTORC1 is not activated during development, but co-incident with elevations in adiposity. We will explore the physiological significance of a positive feedback loop in SREBP1c using genome-edited rats. This key mTORC1 target plays an important role in de novo lipogenesis and the amplification of SREBP1c action by a transcriptional feed-forward circuit has been proposed to be an important component of both diet-induced hepatic steatosis and obesity. By deleting only the relevant SRE at the endogenous Srebf1 locus, we can test the importance of this circuit in a controlled and direct manner. Importantly, these rats will also allow us to separate the direct activation of SREBP1c by mTORC1 and other signals, from the confounding effects of positive feedback. Together these studies will answer fundamental mechanistic questions regarding how mTORC1 and SREBP1c regulate adipogenesis and lipogenesis, providing insights into potential routes of therapeutic intervention for obesity and liver disease.
Glucocorticoids increase liver glycogen levels, but the mechanism and relevance of this process are unknown. By unbiased analyses of multiple transcriptomes using the NURSA platform we have identified PTG, a glycogen-associated protein phosphatase targeting subunit as a novel, glucocorticoid-induced protein. The objective of this proposal is to characterize the nature and relevance of GR/glucocorticoid-dependent induction of PTG expression. To do this we propose to first identify the mechanism by which glucocorticoids result in increased PTG expression, including identification of regulatory elements in the PTG promoter. Second, using PTG knockout mice, we propose to evaluate the relevance of glucocorticoid-dependent induction of PTG on glucose homeostasis. Together these aims will validate a novel nuclear hormone receptor target and establish its relevance in the endocrine control of glucose metabolism.
Glucocorticoids result in adverse metabolic effects including diabetes, NAFLD and weight gain. We have generated preliminary data suggesting that while these risks are present in lean individuals, the consequences are dramatically worse in obese individuals. This proposal aims to identify the molecular and physiological mechanisms that cause these effects by testing effects of obesity on chromatin availability and the role of adipocytes in these effects. Our goal is to identify why and in what tissues obesity causes increased metabolic risk to glucocorticoids.
Peripheral tissues communicate their status to the brain in ways that can modify behavior by engaging higher order processes. Diet and exercise can have potent effects on behaviors, changing appetite, reducing stress, and activating other aspects of the central and peripheral nervous system. There are likely multiple signals that mediate this, but this project will focus on an emerging hormone called GDF15. In particular, we are interested in the impact of moderate to vigorous physical activity and low-carbohydrate diets on the production of this hormone using a combination of mouse and human studies. Long term, this project will study how this hormone responds to changes in diet and exercise to modify behaviors such as food intake, food preference and anxiety.