Dataset Information

Selective galactose culture condition reveals distinct metabolic signatures in pyruvate dehydrogenase and complex I deficient human skin fibroblasts

ABSTRACT:

INTRODUCTION: A decline in mitochondrial function represents a key factor of a large number of inborn errors of metabolism, which lead to an extremely heterogeneous group of disorders.

OBJECTIVES: To gain insight into the biochemical consequences of mitochondrial dysfunction, we performed a metabolic profiling study in human skin fibroblasts using galactose stress medium, which forces cells to rely on mitochondrial metabolism.

METHODS: Fibroblasts from controls, complex I and pyruvate dehydrogenase (PDH) deficient patients were grown under glucose or galactose culture condition. We investigated extracellular flux using Seahorse XF24 cell analyzer and assessed metabolome fingerprints using NMR spectroscopy.

RESULTS: Incubation of fibroblasts in galactose leads to an increase in oxygen consumption and decrease in extracellular acidification rate, confirming adaptation to a more aerobic metabolism. NMR allowed rapid profiling of 41 intracellular metabolites and revealed clear separation of mitochondrial defects from controls under galactose using partial least squares discriminant analysis. We found changes in classical markers of mitochondrial metabolic dysfunction, as well as unexpected markers of amino acid and choline metabolism. PDH deficient cell lines showed distinct upregulation of glutaminolytic metabolism and accumulation of branched-chain amino acids, while complex I deficient cell lines were characterized by increased levels in choline metabolites under galactose.

CONCLUSION: Our results show the relevance of selective culture methods in discriminating normal from metabolic deficient cells. The study indicates that untargeted fingerprinting NMR profiles provide physiological insight on metabolic adaptations and can be used to distinguish cellular metabolic adaptations in PDH and complex I deficient fibroblasts.

INSTRUMENT(S): Nuclear Magnetic Resonance (NMR)

SUBMITTER: Damian Hertig

PROVIDER: MTBLS761 | MetaboLights | 2021-10-06

REPOSITORIES: MetaboLights

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Selective galactose culture condition reveals distinct metabolic signatures in pyruvate dehydrogenase and complex I deficient human skin fibroblasts.

Hertig Damian D Felser Andrea A Diserens Gaëlle G Kurth Sandra S Vermathen Peter P Nuoffer Jean-Marc JM

Metabolomics : Official journal of the Metabolomic Society 20190228 3

<h4>Introduction</h4>A decline in mitochondrial function represents a key factor of a large number of inborn errors of metabolism, which lead to an extremely heterogeneous group of disorders.<h4>Objectives</h4>To gain insight into the biochemical consequences of mitochondrial dysfunction, we performed a metabolic profiling study in human skin fibroblasts using galactose stress medium, which forces cells to rely on mitochondrial metabolism.<h4>Methods</h4>Fibroblasts from controls, complex I and ...[more]

PMID: 30830487

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			Action	DRS
	MTBLS761	Other
	FILES	Other
	a_MTBLS761_NMR_spectroscopy.txt	Txt
	files-all.json	Other
	i_Investigation.txt	Txt