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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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Autores principales: Hannibal, Luciana, Theimer, Jule, Wingert, Victoria, Klotz, Katharina, Bierschenk, Iris, Nitschke, Roland, Spiekerkoetter, Ute, Grünert, Sarah C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7719825/
https://www.ncbi.nlm.nih.gov/pubmed/33329388
http://dx.doi.org/10.3389/fendo.2020.579981
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author Hannibal, Luciana
Theimer, Jule
Wingert, Victoria
Klotz, Katharina
Bierschenk, Iris
Nitschke, Roland
Spiekerkoetter, Ute
Grünert, Sarah C.
author_facet Hannibal, Luciana
Theimer, Jule
Wingert, Victoria
Klotz, Katharina
Bierschenk, Iris
Nitschke, Roland
Spiekerkoetter, Ute
Grünert, Sarah C.
author_sort Hannibal, Luciana
collection PubMed
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.
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spelling pubmed-77198252020-12-15 Metabolic Profiling in Human Fibroblasts Enables Subtype Clustering in Glycogen Storage Disease Hannibal, Luciana Theimer, Jule Wingert, Victoria Klotz, Katharina Bierschenk, Iris Nitschke, Roland Spiekerkoetter, Ute Grünert, Sarah C. Front Endocrinol (Lausanne) Endocrinology 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. Frontiers Media S.A. 2020-11-23 /pmc/articles/PMC7719825/ /pubmed/33329388 http://dx.doi.org/10.3389/fendo.2020.579981 Text en Copyright © 2020 Hannibal, Theimer, Wingert, Klotz, Bierschenk, Nitschke, Spiekerkoetter and Grünert http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Endocrinology
Hannibal, Luciana
Theimer, Jule
Wingert, Victoria
Klotz, Katharina
Bierschenk, Iris
Nitschke, Roland
Spiekerkoetter, Ute
Grünert, Sarah C.
Metabolic Profiling in Human Fibroblasts Enables Subtype Clustering in Glycogen Storage Disease
title 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_short 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
topic Endocrinology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7719825/
https://www.ncbi.nlm.nih.gov/pubmed/33329388
http://dx.doi.org/10.3389/fendo.2020.579981
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