Prof. Marc Hellerstein, M.D. PhD
University of California, Berkeley
Metabolic alterations in primary hepatocytes from mouse CD model and effects of redox manipulations – pilot studies
We propose here a series of pilot studies in primary hepatocytes isolated from a mouse CD-G3PDHm double KO mouse model. We will explore whether ex vivo hepatocyte cultures can reliably quantify consequences on metabolic fluxes and the response to redox-altering interventions in this double-knockout model. The main goal here is to show whether primary hepatocytes can be useful for investigating metabolic alterations and the response to redox-targeted interventions – specifically, do the cells maintain their liver cell phenotype, their CD phenotype and the transfected gene phenotype. Advances in stable isotope -mass spectrometric methods allow key questions to be addressed in vivo, including: measurement of hepatic de novo lipogenesis, gluconeogenesis, glyceroneogenesis, cytosolic acetyl-CoA and triose-phosphate production rates by mass isotopomer distribution analysis; and measurement of hepatic-specific glycolysis/gluconeogenesis, long chain & medium chain FA oxidation by loss of deuterium from 2H-labeled substrates to body water. In addition, a powerful tool to dissipate cellular NADH by expressing bacterial NADH oxidase (LbNOX) will be explored. We hope to demonstrate in these pilot studies that primary hepatocytes can be used to better understand the basic metabolic consequences of CD in the liver.
(Updated November 2023)
Dr. Hellerstein is Professor of Human Nutrition at the University of California, Berkeley, where he occupies an Endowed Chair (Dr. Robert C. and Veronica Atkins Chair). He is also Professor of Endocrinology, Metabolism and Nutrition in the Department of Medicine at the University of California at San Francisco, where he ran the diabetes clinic at SF General Hospital for 25 years. Dr. Hellerstein’s medical training was at Yale Medical School and he completed a PhD at the Massachusetts Institute of Technology, in addition to medical training in Internal Medicine and Endocrinology and Metabolism.