Metabolic Profiling in Human Fibroblasts Enables Subtype Clustering in Glycogen Storage Disease
Glycogen storage disease subtypes I and III (GSD I and GSD III) are monogenic inherited disorders of metabolism that disrupt glycogen metabolism. Unavailability of glucose in GSD I and induction of gluconeogenesis in GSD III modify energy sources and possibly, mitochondrial function. Abnormal mitoch...
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doaj-ca1717f1f4a64b22bae0089ce6cefbf62020-11-25T04:11:49ZengFrontiers Media S.A.Frontiers in Endocrinology1664-23922020-11-011110.3389/fendo.2020.579981579981Metabolic Profiling in Human Fibroblasts Enables Subtype Clustering in Glycogen Storage DiseaseLuciana Hannibal0Jule Theimer1Victoria Wingert2Katharina Klotz3Iris Bierschenk4Roland Nitschke5Roland Nitschke6Ute Spiekerkoetter7Sarah C. Grünert8Laboratory of Clinical Biochemistry and Metabolism, Department of General Pediatrics, Adolescent Medicine and Neonatology, Faculty of Medicine, Medical Center—University of Freiburg, Freiburg, GermanyLaboratory of Clinical Biochemistry and Metabolism, Department of General Pediatrics, Adolescent Medicine and Neonatology, Faculty of Medicine, Medical Center—University of Freiburg, Freiburg, GermanyLaboratory of Clinical Biochemistry and Metabolism, Department of General Pediatrics, Adolescent Medicine and Neonatology, Faculty of Medicine, Medical Center—University of Freiburg, Freiburg, GermanyLaboratory of Clinical Biochemistry and Metabolism, Department of General Pediatrics, Adolescent Medicine and Neonatology, Faculty of Medicine, Medical Center—University of Freiburg, Freiburg, GermanyLife Imaging Center, Center for Integrated Signalling Analysis, Albert-Ludwigs-University, Freiburg, GermanyLife Imaging Center, Center for Integrated Signalling Analysis, Albert-Ludwigs-University, Freiburg, GermanyBIOSS Centre for Biological Signaling Studies, Albert-Ludwigs-University Freiburg, Freiburg, GermanyDepartment of General Pediatrics, Adolescent Medicine and Neonatology, Faculty of Medicine, Medical Center—University of Freiburg, Freiburg, GermanyDepartment of General Pediatrics, Adolescent Medicine and Neonatology, Faculty of Medicine, Medical Center—University of Freiburg, Freiburg, GermanyGlycogen storage disease subtypes I and III (GSD I and GSD III) are monogenic inherited disorders of metabolism that disrupt glycogen metabolism. Unavailability of glucose in GSD I and induction of gluconeogenesis in GSD III modify energy sources and possibly, mitochondrial function. Abnormal mitochondrial structure and function were described in mice with GSD Ia, yet significantly less research is available in human cells and ketotic forms of the disease. We hypothesized that impaired glycogen storage results in distinct metabolic phenotypes in the extra- and intracellular compartments that may contribute to pathogenesis. Herein, we examined mitochondrial organization in live cells by spinning-disk confocal microscopy and profiled extra- and intracellular metabolites by targeted LC-MS/MS in cultured fibroblasts from healthy controls and from patients with GSD Ia, GSD Ib, and GSD III. Results from live imaging revealed that mitochondrial content and network morphology of GSD cells are comparable to that of healthy controls. Likewise, healthy controls and GSD cells exhibited comparable basal oxygen consumption rates. Targeted metabolomics followed by principal component analysis (PCA) and hierarchical clustering (HC) uncovered metabolically distinct poises of healthy controls and GSD subtypes. Assessment of individual metabolites recapitulated dysfunctional energy production (glycolysis, Krebs cycle, succinate), reduced creatinine export in GSD Ia and GSD III, and reduced antioxidant defense of the cysteine and glutathione systems. Our study serves as proof-of-concept that extra- and intracellular metabolite profiles distinguish glycogen storage disease subtypes from healthy controls. We posit that metabolite profiles provide hints to disease mechanisms as well as to nutritional and pharmacological elements that may optimize current treatment strategies.https://www.frontiersin.org/articles/10.3389/fendo.2020.579981/fullinborn error of metabolismmetabolismmetabolomicsmitochondriaglycogen storage diseaseenergy deficiency |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Luciana Hannibal Jule Theimer Victoria Wingert Katharina Klotz Iris Bierschenk Roland Nitschke Roland Nitschke Ute Spiekerkoetter Sarah C. Grünert |
spellingShingle |
Luciana Hannibal Jule Theimer Victoria Wingert Katharina Klotz Iris Bierschenk Roland Nitschke Roland Nitschke Ute Spiekerkoetter Sarah C. Grünert Metabolic Profiling in Human Fibroblasts Enables Subtype Clustering in Glycogen Storage Disease Frontiers in Endocrinology inborn error of metabolism metabolism metabolomics mitochondria glycogen storage disease energy deficiency |
author_facet |
Luciana Hannibal Jule Theimer Victoria Wingert Katharina Klotz Iris Bierschenk Roland Nitschke Roland Nitschke Ute Spiekerkoetter Sarah C. Grünert |
author_sort |
Luciana Hannibal |
title |
Metabolic Profiling in Human Fibroblasts Enables Subtype Clustering in Glycogen Storage Disease |
title_short |
Metabolic Profiling in Human Fibroblasts Enables Subtype Clustering in Glycogen Storage Disease |
title_full |
Metabolic Profiling in Human Fibroblasts Enables Subtype Clustering in Glycogen Storage Disease |
title_fullStr |
Metabolic Profiling in Human Fibroblasts Enables Subtype Clustering in Glycogen Storage Disease |
title_full_unstemmed |
Metabolic Profiling in Human Fibroblasts Enables Subtype Clustering in Glycogen Storage Disease |
title_sort |
metabolic profiling in human fibroblasts enables subtype clustering in glycogen storage disease |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Endocrinology |
issn |
1664-2392 |
publishDate |
2020-11-01 |
description |
Glycogen storage disease subtypes I and III (GSD I and GSD III) are monogenic inherited disorders of metabolism that disrupt glycogen metabolism. Unavailability of glucose in GSD I and induction of gluconeogenesis in GSD III modify energy sources and possibly, mitochondrial function. Abnormal mitochondrial structure and function were described in mice with GSD Ia, yet significantly less research is available in human cells and ketotic forms of the disease. We hypothesized that impaired glycogen storage results in distinct metabolic phenotypes in the extra- and intracellular compartments that may contribute to pathogenesis. Herein, we examined mitochondrial organization in live cells by spinning-disk confocal microscopy and profiled extra- and intracellular metabolites by targeted LC-MS/MS in cultured fibroblasts from healthy controls and from patients with GSD Ia, GSD Ib, and GSD III. Results from live imaging revealed that mitochondrial content and network morphology of GSD cells are comparable to that of healthy controls. Likewise, healthy controls and GSD cells exhibited comparable basal oxygen consumption rates. Targeted metabolomics followed by principal component analysis (PCA) and hierarchical clustering (HC) uncovered metabolically distinct poises of healthy controls and GSD subtypes. Assessment of individual metabolites recapitulated dysfunctional energy production (glycolysis, Krebs cycle, succinate), reduced creatinine export in GSD Ia and GSD III, and reduced antioxidant defense of the cysteine and glutathione systems. Our study serves as proof-of-concept that extra- and intracellular metabolite profiles distinguish glycogen storage disease subtypes from healthy controls. We posit that metabolite profiles provide hints to disease mechanisms as well as to nutritional and pharmacological elements that may optimize current treatment strategies. |
topic |
inborn error of metabolism metabolism metabolomics mitochondria glycogen storage disease energy deficiency |
url |
https://www.frontiersin.org/articles/10.3389/fendo.2020.579981/full |
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