DC15

Background: In computational, animal and cell culture models of a fatty acid oxidation disorder (medium-chain acyl-CoA dehydrogenase deficiency (MCADD)), we observed a substantial sequestration of CoA in acyl-CoA esters and a concomitant decrease of the free CoA that is available for other metabolic processes. When cells/mice are dependent on fatty-acid oxidation (e.g., cold or low glucose), they exhibit a massive compensatory response in CoA biosynthesis and in pathways liberating free CoA. We hypothesize that the nutritional status also plays a role in primary CoA biosynthesis defects, such as PKAN. To study the functional impact of MCADD and PKAN on the dynamics of CoA and acyl-CoA esters of all chain lengths, we will use stable-isotope precursors, HILIC-MS-MS methodology and mild extraction leaving CoA esters intact. We noticed previously that this ‘metabolically active’ CoA pool is labelled faster than the total CoA pool.

Objectives: (1) Quantify the functional impact of MCADD and PKAN on CoA biosynthesis by isotope labelling and HILIC-MS-MS in MCADD and PKAN cell models; (2) Assess the impact of available nutrients (glucose, fatty acid, carnitine) on CoA dynamics in these disease models (3) Develop a computational model of isotope dynamics to infer fluxes of (acyl)-CoA metabolism in MCADD and PKAN.