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Pilot & Feasibility Program Application Abstract

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Quantifying the reaction rates that affect fatty liver and hepatic function

Michelle Puchowicz (Memphis, TN)

Pilot & Feasibility Program

Investigators have documented a clear relationship between hepatic lipid content and the progression towards liver failure and related disease(s) (e.g. diabetes), however, there is limited information regarding the biochemical flux rates that affect fatty liver and the consequences of fatty liver on liver function. Our primary aim is to develop a novel stable isotope tracer method (that relies on the administration of D2O) for quantifying the rate of hepatic triglyceride synthesis/secretion and to determine the contributions of de novo lipogenesis and reesterification to the source of fatty acids. Our secondary aim is to quantify the effect(s) of fatty liver on two critical hepatic functions (managing glucose flux and albumin synthesis/secretion) which can be determined using the same D2O method. We plan to utilize experimental models including C57BL/6J mice fed either a low-fat (LF) or a high-fat (HF) diet and studied during basal or glucose-insulin clamp conditions (hyperglycemic-hyperinsulinemic) and PPAR-alpha KO mice during short vs. prolonged fasting in the presence or absence of acipimox treatment (biological studies will be done in the Vanderbilt MMPC and analyses will be done in the CWRU MMPC). These physiological conditions should allow us to make reasonable predictions regarding the importance of specific reactions a priori and therein provide a rigorous test of the method, e.g. we expect and increase in triglyceride synthesis in HF vs. LF fed mice, we expect an increased contribution of de novo lipogenesis and a reduction in the contribution of reesterification in basal vs. "clamped" mice, etc. Successful completion of our studies will (i) generate a new tool for MMPC users regarding lipid flux and liver disease, (ii) demonstrate the importance and potential of inter-MMPC activities and (iii) generate data to be used in a larger grant that is aimed at examining the efficacy of different therapeutic interventions for managing fatty liver and adapting these methods for use in humans.

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