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

Hertig, Damian; Felser, Andrea; Diserens, Gaëlle; Kurth, Sandra; Vermathen, Peter; Nuoffer, Jean-Marc (2019). Selective galactose culture condition reveals distinct metabolic signatures in pyruvate dehydrogenase and complex I deficient human skin fibroblasts. Metabolomics, 15(3), p. 32. Springer 10.1007/s11306-019-1497-2

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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.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Pavillon 52 > Abt. Magnetresonanz-Spektroskopie und Methodologie, AMSM
04 Faculty of Medicine > Department of Haematology, Oncology, Infectious Diseases, Laboratory Medicine and Hospital Pharmacy (DOLS) > Institute of Clinical Chemistry
04 Faculty of Medicine > Department of Gynaecology, Paediatrics and Endocrinology (DFKE) > Clinic of Paediatric Medicine

UniBE Contributor:

Felser, Andrea; Diserens, Gaëlle; Kurth, Sandra; Vermathen, Peter and Nuoffer, Jean-Marc

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1573-3882

Publisher:

Springer

Language:

English

Submitter:

Maria de Fatima Henriques Bernardo

Date Deposited:

25 Apr 2019 10:40

Last Modified:

23 Oct 2019 07:54

Publisher DOI:

10.1007/s11306-019-1497-2

Related URLs:

PubMed ID:

30830487

Uncontrolled Keywords:

Complex I Galactose Mitochondrial dysfunction NMR Pyruvate dehydrogenase

BORIS DOI:

10.7892/boris.127461

URI:

https://boris.unibe.ch/id/eprint/127461

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