Improved detection of long‐chain fatty acid oxidation defects in intact cells using [9,10‐3H]oleic acid

Olpin, S. E.; Manning, N. J.; Pollitt, R. J.; Clarke, Steven D.

doi:10.1023/a:1005358802096

Cited by 56 publications

(16 citation statements)

References 6 publications

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“…Most of the lcFAODs result in a distinctive acylcarnitine profile depending on which of the lcFAO enzymes is deficient (Table 1 ). Overall lcFAO activity can also be measured in fibroblasts; a whole cell assay, using radiolabeled fatty acids as a substrate [ 48 , 49 ]. In addition, the activity of specific enzymes can be measured in lymphocytes or in skin fibroblasts, as these cell types express all lcFAO enzymes [ 5 ].…”

Section: Diagnostic Approach For Long-chain Fatty Acid Oxidation Disomentioning

confidence: 99%

Disorders of mitochondrial long-chain fatty acid oxidation and the carnitine shuttle

Knottnerus

Bleeker

Wüst

et al. 2018

Rev Endocr Metab Disord

234

209

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Mitochondrial fatty acid oxidation is an essential pathway for energy production, especially during prolonged fasting and sub-maximal exercise. Long-chain fatty acids are the most abundant fatty acids in the human diet and in body stores, and more than 15 enzymes are involved in long-chain fatty acid oxidation. Pathogenic mutations in genes encoding these enzymes result in a long-chain fatty acid oxidation disorder in which the energy homeostasis is compromised and long-chain acylcarnitines accumulate. Symptoms arise or exacerbate during catabolic situations, such as fasting, illness and (endurance) exercise. The clinical spectrum is very heterogeneous, ranging from hypoketotic hypoglycemia, liver dysfunction, rhabdomyolysis, cardiomyopathy and early demise. With the introduction of several of the long-chain fatty acid oxidation disorders (lcFAOD) in newborn screening panels, also asymptomatic individuals with a lcFAOD are identified. However, despite early diagnosis and dietary therapy, a significant number of patients still develop symptoms emphasizing the need for individualized treatment strategies. This review aims to function as a comprehensive reference for clinical and laboratory findings for clinicians who are confronted with pediatric and adult patients with a possible diagnosis of a lcFAOD.

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Section: Diagnostic Approach For Long-chain Fatty Acid Oxidation Disomentioning

confidence: 99%

Disorders of mitochondrial long-chain fatty acid oxidation and the carnitine shuttle

Knottnerus

Bleeker

Wüst

et al. 2018

Rev Endocr Metab Disord

234

209

View full text Add to dashboard Cite

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“…Skin fibroblast were plated in 48-well plates. Long-chain fatty acid oxidation was measured by the production of 3 H 2 O from [9-3 H(N)]-oleic acid as described previously [45]. Measurements were done at 37 • C in duplicate and lcFAO flux is expressed as percentage of mean activity of skin fibroblasts of healthy controls measured in the same experiment.…”

Section: Measurement Of Long-chain Fatty Acid Oxidation (Lcfao) Fluxmentioning

confidence: 99%

Electrophysiological Abnormalities in VLCAD Deficient hiPSC-Cardiomyocytes Can Be Improved by Lowering Accumulation of Fatty Acid Oxidation Intermediates

Knottnerus

Mengarelli

Wüst

et al. 2020

IJMS

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Patients with very long-chain acyl-CoA dehydrogenase deficiency (VLCADD) can present with life-threatening cardiac arrhythmias. The pathophysiological mechanism is unknown. We reprogrammed fibroblasts from one mildly and one severely affected VLCADD patient, into human induced pluripotent stem cells (hiPSCs) and differentiated these into cardiomyocytes (VLCADD-CMs). VLCADD-CMs displayed shorter action potentials (APs), more delayed afterdepolarizations (DADs) and higher systolic and diastolic intracellular Ca2+ concentration ([Ca2+]i) than control CMs. The mitochondrial booster resveratrol mitigated the biochemical, electrophysiological and [Ca2+]i changes in the mild but not in the severe VLCADD-CMs. Accumulation of potentially toxic intermediates of fatty acid oxidation was blocked by substrate reduction with etomoxir. Incubation with etomoxir led to marked prolongation of AP duration and reduced DADs and [Ca2+]i in both VLCADD-CMs. These results provide compelling evidence that reduced accumulation of fatty acid oxidation intermediates, either by enhanced fatty acid oxidation flux through increased mitochondria biogenesis (resveratrol) or by inhibition of fatty acid transport into the mitochondria (etomoxir), rescues pro-arrhythmia defects in VLCADD-CMs and open doors for new treatments.

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“…Through the years, many different methods have been set up for this purpose, including: (1) [ 14 C]-CO 2 release assays using different [1- 14 C]- or [U- 14 C]-labelled fatty acids; (2) tritium release assays using different [ 3 H]-labelled fatty acids, notably [9,10- 3 H]-myristic acid, [9,10- 3 H]-palmitic acid, and [9,10- 3 H]-oleic acid (Manning et al 1990; Olpin et al 1992, 1997); and (3) quantitative acyl-CoA and acylcarnitine profiling studies. The latter methods have been pioneered by Bartlett and co-workers in the late 1980s during studies on the beta-oxidation of fatty acids in rat liver mitochondria.…”

Section: The Mitochondrial Fatty Acyl-coa Oxidation Systemmentioning

confidence: 99%

The enzymology of mitochondrial fatty acid beta‐oxidation and its application to follow‐up analysis of positive neonatal screening results

Wanders

Ruiter

IJlst

et al. 2010

J of Inher Metab Disea

177

147

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Oxidation of fatty acids in mitochondria is a key physiological process in higher eukaryotes including humans. The importance of the mitochondrial beta-oxidation system in humans is exemplified by the existence of a group of genetic diseases in man caused by an impairment in the mitochondrial oxidation of fatty acids. Identification of patients with a defect in mitochondrial beta-oxidation has long remained notoriously difficult, but the introduction of tandem-mass spectrometry in laboratories for genetic metabolic diseases has revolutionalized the field by allowing the rapid and sensitive analysis of acylcarnitines. Equally important is that much progress has been made with respect to the development of specific enzyme assays to identify the enzyme defect in patients subsequently followed by genetic analysis. In this review, we will describe the current state of knowledge in the field of fatty acid oxidation enzymology and its application to the follow-up analysis of positive neonatal screening results.

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Improved detection of long‐chain fatty acid oxidation defects in intact cells using [9,10‐3H]oleic acid

Cited by 56 publications

References 6 publications

Disorders of mitochondrial long-chain fatty acid oxidation and the carnitine shuttle

Disorders of mitochondrial long-chain fatty acid oxidation and the carnitine shuttle

Electrophysiological Abnormalities in VLCAD Deficient hiPSC-Cardiomyocytes Can Be Improved by Lowering Accumulation of Fatty Acid Oxidation Intermediates

The enzymology of mitochondrial fatty acid beta‐oxidation and its application to follow‐up analysis of positive neonatal screening results

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