Mechanisms of Dietary Restriction mediated by Creb-binding Protein in Mice

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DESCRIPTION (provided by applicant): Dietary Restriction is a manipulation that reduces caloric intake and subsequently extends lifespan and protects against age-related diseases in animal models. The protective spectrum of this manipulation includes prevalent diseases such as cancer, diabetes, cardiovascular disease, and Alzheimer's disease. It is therefore imperative to understand the underlying metabolic mechanisms of this paradigm. CBP is a transcriptional regulator that has been implicated in both the protective and age-extending properties of dietary restriction. Of importance, evidence suggests CBP exerts these effects by mainly acting in the hypothalamus, a sensory center of metabolic state. The purpose of these studies is to study the role of CBP and the metabolic profile of dietary restriction with an emphasis in hypothalamic responses. To carry these studies we will develop mice that lack CBP post-natally. These mice will then be subjected to either acute (8 weeks) or chronic (18 month) dietary restriction. Following this period, relevant tissues will be collected and analyzed for metabolic profiles. Specifically, we will 1) monitor metabolic gene expression changes using custom qPCR plates; 2) observe metabolic gene targeting changes of CBP using Chromatin immunoprecipitation followed by RT-PCR; 3) quantify metabolite usage by liquid chromatography followed by mass spectrometry. These assays will be performed in both chronic and acute groups. Additionally, we will assess the chronically dietary restricted mice using genetic markers of senescence, which will help discern if the protective effects are attributable to CBP expression in the hypothalamus. Other neuroendocrine responses will be monitored for all groups further characterizing the physiology of this dietary manipulation. Upon completion of this project it is expected that a clearer metabolic and transcriptional profile of dietary restriction will be established, taking us one step closer to the development of clinical applications. PUBLIC HEALTH RELEVANCE: Dietary Restriction is currently the only manipulation that repeatedly results in both lifespan extension and age-related disease protection across species. It is therefore of clinical relevance to study these complex mechanisms in hope to mimic these effects in the future. This proposal intends to elucidate the mechanisms by which a transcriptional factor, CBP, mediates these effects.