Prof. Marc Hellerstein, M.D. PhD
University of California, Berkeley

Metabolic alterations in vivo in liver of rodent CD double knock-out model and effects of redox manipulations

CD is arguably the cleanest example of a liver redox disease in human biology. Yet the metabolic consequences – the phenotype – of this monogenic disorder remain poorly understood. Although human CD has some predictable metabolic consequences, many metabolic features defy current knowledge about hepatic metabolic regulation.

We propose studies to quantify metabolic flux alterations fluxes in liver in vivo in a mouse CD model (citrin-G3PDHm double knockout) and the response to interventions (milestone-based, in year 2). Advances in stable isotope-mass spectrometric methods allow basic metabolic control questions to be measured, including: hepatic de novo lipogenesis, gluconeogenesis, glyceroneogenesis and production rates of cytosolic acetyl-CoA and triose-phosphate, by mass isotopomer distribution analysis; hepatic-specific glycolysis/gluconeogenesis and oxidation of long chain & medium chain fatty acids by loss of deuterium from 2H-labeled substrates; and synthesis of key liver enzymes, including arginosuccinate synthetase, by heavy water labeling. In addition, a powerful tool to dissipate cellular NADH through expression of bacterial NADH oxidase (LbNOX) will be deployed and the therapeutic consequences of medium chain triglyceride (MCT) administration will be established. Our primary objectives are to correlate measurements of key hepatic metabolic pathway fluxes with molecular changes (gene expression, redox state) and to test interventions on cytosolic redox (LbNOX, ethanol) or lipid substrates (MCT). We propose a milestone-based decision for advancing to year 2 or potentially switching to a rat CD model. In summary, we hope to understand basic metabolic consequences of CD in liver, to promote effective management and development of new treatments for CD.

(Updated November 2023)