(282c) 13C Flux Analysis in Metabolism Research: From Cells to Multi-Tissue In Vivo Models

¹³C metabolic flux analysis (MFA) is the gold standard approach for quantifying rates of biochemical reactions in living cells and tissues. It has been widely applied to assess the metabolic phenotypes of industrial host organisms, but it is now being increasingly used to investigate metabolic disease mechanisms both in cellular and in vivo models. Over the past 15 years, my lab has established novel ¹³C MFA tools and approaches that enable us to probe entirely new aspects of metabolism previously inaccessible to measurement. In particular, we have developed publicly available software packages PIRAMID and INCA that automate the computational workflows of ¹³C tracer analysis and data modeling. I will discuss ongoing work where my lab has leveraged these tools to investigate the pathogenesis of metabolic dysfunction-associated steatotic liver disease (MASLD) using cultured cells and mouse models. We have recently expanded our in vivo ¹³C MFA platform to enable simultaneous determination of intermediary metabolic fluxes in both glycolytic (e.g., muscle, adipose) and gluconeogenic tissues (e.g., liver) of individual mice based on modeling metabolomics datasets obtained from infusion of stable isotopes. These studies have established ¹³C flux analysis and the INCA software suite as a comprehensive platform to investigate intermediary metabolism in a wide range of biological contexts.