FATTY-ACID OXIDATION IN PERIPHERAL-BLOOD CELLS - CHARACTERIZATION AND USE FOR THE DIAGNOSIS OF DEFECTS OF FATTY-ACID OXIDATION

Disorders of mitochondrial fatty acid oxidation are increasingly recognized as an important group of inborn errors of metabolism that are associated with a significant, but easily preventable, morbidity and mortality in children. However, diagnosis is often delayed because there is no easily applied method that detects all defects. Therefore, we have characterized the acylcarnitine intermediates of fatty acid oxidation in peripheral blood cells from healthy control volunteers and patients with four different defects. After selective permeabilization with histone II AS, a novel permeabilizing agent, the cells were incubated with [U-C-14]hexadecanoate and beta-oxidation flux and the acylcarnitine esters formed were measured. Blood cells from the control population produced large amounts of 3-hydroxyacylcarnitines and 2-enoylcarnitine esters, in addition to saturated acylcarnitine esters. This result is different from that found in other tissues (fibroblasts and muscle), where only saturated acylcarnitine esters could be detected. In blood cells from patients with defects of enzymes involved in long-chain fatty acid oxidation, flux was significantly reduced at 15 to 20% of control values (7.1 +/- 2.3 nmol C-2 units formed per minute per International Unit of citrate synthase activity). There was a characteristic accumulation of acylcarnitines that was pathognomonic for the site of the defect. Thus, analysis of beta-oxidation intermediates from blood cells allows unequivocal identification of the four most common beta-oxidation defects.