{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T16:53:56Z","timestamp":1790787236776,"version":"4.1.0"},"reference-count":28,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2023,5,4]],"date-time":"2023-05-04T00:00:00Z","timestamp":1683158400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Netherlands Organization for Scientific Research","award":["91713359"],"award-info":[{"award-number":["91713359"]}]},{"name":"Netherlands Organization for Scientific Research","award":["09150182010010"],"award-info":[{"award-number":["09150182010010"]}]},{"name":"Netherlands Organization for Scientific Research","award":["W.OR17-15"],"award-info":[{"award-number":["W.OR17-15"]}]},{"name":"Netherlands Organization for Scientific Research","award":["2019-2766-001"],"award-info":[{"award-number":["2019-2766-001"]}]},{"name":"Netherlands Organization for Scientific Research","award":["UMD-CG-2020-006"],"award-info":[{"award-number":["UMD-CG-2020-006"]}]},{"name":"Netherlands X-omics Initiative","award":["91713359"],"award-info":[{"award-number":["91713359"]}]},{"name":"Netherlands X-omics Initiative","award":["09150182010010"],"award-info":[{"award-number":["09150182010010"]}]},{"name":"Netherlands X-omics Initiative","award":["W.OR17-15"],"award-info":[{"award-number":["W.OR17-15"]}]},{"name":"Netherlands X-omics Initiative","award":["2019-2766-001"],"award-info":[{"award-number":["2019-2766-001"]}]},{"name":"Netherlands X-omics Initiative","award":["UMD-CG-2020-006"],"award-info":[{"award-number":["UMD-CG-2020-006"]}]},{"name":"Stichting Stofwisselkracht","award":["91713359"],"award-info":[{"award-number":["91713359"]}]},{"name":"Stichting Stofwisselkracht","award":["09150182010010"],"award-info":[{"award-number":["09150182010010"]}]},{"name":"Stichting Stofwisselkracht","award":["W.OR17-15"],"award-info":[{"award-number":["W.OR17-15"]}]},{"name":"Stichting Stofwisselkracht","award":["2019-2766-001"],"award-info":[{"award-number":["2019-2766-001"]}]},{"name":"Stichting Stofwisselkracht","award":["UMD-CG-2020-006"],"award-info":[{"award-number":["UMD-CG-2020-006"]}]},{"name":"UMD","award":["91713359"],"award-info":[{"award-number":["91713359"]}]},{"name":"UMD","award":["09150182010010"],"award-info":[{"award-number":["09150182010010"]}]},{"name":"UMD","award":["W.OR17-15"],"award-info":[{"award-number":["W.OR17-15"]}]},{"name":"UMD","award":["2019-2766-001"],"award-info":[{"award-number":["2019-2766-001"]}]},{"name":"UMD","award":["UMD-CG-2020-006"],"award-info":[{"award-number":["UMD-CG-2020-006"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Phosphoglucomutase 1 (PGM1) is a key enzyme for the regulation of energy metabolism from glycogen and glycolysis, as it catalyzes the interconversion of glucose 1-phosphate and glucose 6-phosphate. PGM1 deficiency is an autosomal recessive disorder characterized by a highly heterogenous clinical spectrum, including hypoglycemia, cleft palate, liver dysfunction, growth delay, exercise intolerance, and dilated cardiomyopathy. Abnormal protein glycosylation has been observed in this disease. Oral supplementation with D-galactose efficiently restores protein glycosylation by replenishing the lacking pool of UDP-galactose, and rescues some symptoms, such as hypoglycemia, hepatopathy, and growth delay. However, D-galactose effects on skeletal muscle and heart symptoms remain unclear. In this study, we established an in vitro muscle model for PGM1 deficiency to investigate the role of PGM1 and the effect of D-galactose on nucleotide sugars and energy metabolism. Genome-editing of C2C12 myoblasts via CRISPR\/Cas9 resulted in Pgm1 (mouse homologue of human PGM1, according to updated nomenclature) knockout clones, which showed impaired maturation to myotubes. No difference was found for steady-state levels of nucleotide sugars, while dynamic flux analysis based on 13C6-galactose suggested a block in the use of galactose for energy production in knockout myoblasts. Subsequent analyses revealed a lower basal respiration and mitochondrial ATP production capacity in the knockout myoblasts and myotubes, which were not restored by D-galactose. In conclusion, an in vitro mouse muscle cell model has been established to study the muscle-specific metabolic mechanisms in PGM1 deficiency, which suggested that galactose was unable to restore the reduced energy production capacity.<\/jats:p>","DOI":"10.3390\/ijms24098247","type":"journal-article","created":{"date-parts":[[2023,5,5]],"date-time":"2023-05-05T03:24:32Z","timestamp":1683257072000},"page":"8247","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["In Vitro Skeletal Muscle Model of PGM1 Deficiency Reveals Altered Energy Homeostasis"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1481-7401","authenticated-orcid":false,"given":"Federica","family":"Conte","sequence":"first","affiliation":[{"name":"Department of Neurology, Donders Institute for Brain, Cognition and Behavior, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9679-7955","authenticated-orcid":false,"given":"Angel","family":"Ashikov","sequence":"additional","affiliation":[{"name":"Department of Neurology, Donders Institute for Brain, Cognition and Behavior, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0311-6482","authenticated-orcid":false,"given":"Rachel","family":"Mijdam","sequence":"additional","affiliation":[{"name":"Department of Neurology, Donders Institute for Brain, Cognition and Behavior, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eline G. P.","family":"van de Ven","sequence":"additional","affiliation":[{"name":"Department of Neurology, Donders Institute for Brain, Cognition and Behavior, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monique","family":"van Scherpenzeel","sequence":"additional","affiliation":[{"name":"GlycoMScan B.V., 5349AB Oss, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raisa","family":"Veizaj","sequence":"additional","affiliation":[{"name":"Department of Neurology, Donders Institute for Brain, Cognition and Behavior, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seyed P.","family":"Mahalleh-Yousefi","sequence":"additional","affiliation":[{"name":"Department of Neurology, Donders Institute for Brain, Cognition and Behavior, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7433-9048","authenticated-orcid":false,"given":"Merel A.","family":"Post","sequence":"additional","affiliation":[{"name":"Department of Neurology, Donders Institute for Brain, Cognition and Behavior, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karin","family":"Huijben","sequence":"additional","affiliation":[{"name":"Translational Metabolic Laboratory, Department of Laboratory Medicine, Radboud Institute for Molecular Life Sciences (RIMLS), Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daan M.","family":"Panneman","sequence":"additional","affiliation":[{"name":"Radboud Center for Mitochondrial Medicine (RCMM), Department of Pediatrics, Amalia Children\u2019s Hospital, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"},{"name":"Radboud Institute for Molecular Life Sciences (RIMLS), Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5227-3527","authenticated-orcid":false,"given":"Richard J. T.","family":"Rodenburg","sequence":"additional","affiliation":[{"name":"Translational Metabolic Laboratory, Department of Laboratory Medicine, Radboud Institute for Molecular Life Sciences (RIMLS), Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"},{"name":"Radboud Center for Mitochondrial Medicine (RCMM), Department of Pediatrics, Amalia Children\u2019s Hospital, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicol C.","family":"Voermans","sequence":"additional","affiliation":[{"name":"Department of Neurology, Donders Institute for Brain, Cognition and Behavior, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5721-1560","authenticated-orcid":false,"given":"Alejandro","family":"Garanto","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"},{"name":"Department of Pediatrics, Amalia Children\u2019s Hospital, Radboud Institute for Molecular Life Sciences (RIMLS), Radboud University Medical Center, 6525GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Werner J. 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Engl. J. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"835","DOI":"10.1016\/j.ajhg.2019.03.003","article-title":"The Metabolic Map into the Pathomechanism and Treatment of PGM1-CDG","volume":"104","author":"Radenkovic","year":"2019","journal-title":"Am. J. Hum. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1093\/glycob\/cwab106","article-title":"Dynamic tracing of sugar metabolism reveals the mechanisms of action of synthetic sugar analogs","volume":"32","author":"Conte","year":"2022","journal-title":"Glycobiology"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Elkalaf, M., And\u011bl, M., and Trnka, J. (2013). Low glucose but not galactose enhances oxidative mitochondrial metabolism in C2C12 myoblasts and myotubes. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0070772"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3349","DOI":"10.1113\/jphysiol.2012.230185","article-title":"Biomarkers of mitochondrial content in skeletal muscle of healthy young human subjects","volume":"590","author":"Larsen","year":"2012","journal-title":"J. Physiol."},{"key":"ref_6","first-page":"84","article-title":"The relationship between skeletal muscle mitochondrial citrate synthase activity and whole body oxygen uptake adaptations in response to exercise training","volume":"6","author":"Andersen","year":"2014","journal-title":"Int. J. Physiol. Pathophysiol. Pharmacol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"598","DOI":"10.1016\/j.bbabio.2011.07.001","article-title":"Regulation of mitochondrial respiration and apoptosis through cell signaling: Cytochrome c oxidase and cytochrome c in ischemia\/reperfusion injury and inflammation","volume":"1817","author":"Helling","year":"2012","journal-title":"Biochim. Biophys. Acta (BBA) Bioenerg."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"13546","DOI":"10.1073\/pnas.1107553108","article-title":"Nanoscale distribution of mitochondrial import receptor Tom20 is adjusted to cellular conditions and exhibits an inner-cellular gradient","volume":"108","author":"Wurm","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1042\/EBC20170098","article-title":"Assembly of mammalian oxidative phosphorylation complexes I-V and supercomplexes","volume":"62","author":"Signes","year":"2018","journal-title":"Essays Biochem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1016\/B978-0-12-801415-8.00016-3","article-title":"Analysis and interpretation of microplate-based oxygen consumption and pH data","volume":"547","author":"Divakaruni","year":"2014","journal-title":"Methods Enzymol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Y\u00e9pez, V.A., Kremer, L.S., Iuso, A., Gusic, M., Kopajtich, R., Ko\u0148a\u0159\u00edkov\u00e1, E., Nadel, A., Wachutka, L., Prokisch, H., and Gagneur, J. (2018). OCR-Stats: Robust estimation and statistical testing of mitochondrial respiration activities using Seahorse XF Analyzer. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0199938"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"13106","DOI":"10.1096\/fj.202000767R","article-title":"Mitochondrial spare respiratory capacity: Mechanisms, regulation, and significance in non-transformed and cancer cells","volume":"34","author":"Marchetti","year":"2020","journal-title":"FASEB J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1080\/15548627.2015.1115172","article-title":"Mitophagy is required for mitochondrial biogenesis and myogenic differentiation of C2C12 myoblasts","volume":"12","author":"Sin","year":"2016","journal-title":"Autophagy"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"737","DOI":"10.1038\/s41598-017-18658-3","article-title":"The effect of differentiation and TGF\u03b2 on mitochondrial respiration and mitochondrial enzyme abundance in cultured primary human skeletal muscle cells","volume":"8","author":"Hoffmann","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"18232","DOI":"10.1038\/s41598-020-75406-w","article-title":"Substrate utilisation of cultured skeletal muscle cells in patients with CFS","volume":"10","author":"Tomas","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Aguer, C., Gambarotta, D., Mailloux, R.J., Moffat, C., Dent, R., McPherson, R., and Harper, M.E. (2011). Galactose enhances oxidative metabolism and reveals mitochondrial dysfunction in human primary muscle cells. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0028536"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"619710","DOI":"10.3389\/fphys.2021.619710","article-title":"Tissue-Engineered Skeletal Muscle Models to Study Muscle Function, Plasticity, and Disease","volume":"12","author":"Khodabukus","year":"2021","journal-title":"Front. Physiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1093\/toxsci\/kfm052","article-title":"Circumventing the Crabtree effect: Replacing media glucose with galactose increases susceptibility of HepG2 cells to mitochondrial toxicants","volume":"97","author":"Marroquin","year":"2007","journal-title":"Toxicol. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/j.stem.2014.12.004","article-title":"The NAD(+)-dependent SIRT1 deacetylase translates a metabolic switch into regulatory epigenetics in skeletal muscle stem cells","volume":"16","author":"Ryall","year":"2015","journal-title":"Cell Stem Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1016\/0014-5793(95)01357-1","article-title":"Phosphomannomutase deficiency is a cause of carbohydrate-deficient glycoprotein syndrome type I","volume":"377","author":"Jaeken","year":"1995","journal-title":"FEBS Lett."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1666","DOI":"10.1016\/j.bbapap.2009.10.022","article-title":"SIRT1-dependent regulation of chromatin and transcription: Linking NAD(+) metabolism and signaling to the control of cellular functions","volume":"1804","author":"Zhang","year":"2010","journal-title":"Biochim. Biophys. Acta (BBA)-Proteins Proteom."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.ymgme.2020.08.003","article-title":"Phosphoglucomutase-1 deficiency: Early presentation, metabolic management and detection in neonatal blood spots","volume":"131","author":"Conte","year":"2020","journal-title":"Mol. Genet. Metab."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1016\/j.nmd.2017.01.014","article-title":"PGM1 deficiency: Substrate use during exercise and effect of treatment with galactose","volume":"27","author":"Voermans","year":"2017","journal-title":"Neuromuscul. Disord."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.ymgme.2017.08.007","article-title":"Impaired glycogen breakdown and synthesis in phosphoglucomutase 1 deficiency","volume":"122","author":"Preisler","year":"2017","journal-title":"Mol. Genet. Metab."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2281","DOI":"10.1038\/nprot.2013.143","article-title":"Genome engineering using the CRISPR-Cas9 system","volume":"8","author":"Ran","year":"2013","journal-title":"Nat. Protoc."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212\u0394\u0394CT Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_27","unstructured":"Conte, F.N. (2023). Phosphoglucomutase 1 Deficiency: From Clinical Presentation To Sugar Metabolism In Cell Models. [Ph.D. Thesis, Radboud University]. Chapter 5."},{"key":"ref_28","unstructured":"Panneman, D.M. (2021). Exploring Novel Mechanisms Causing Mitochondrial Disorders through Functional Genomics. [Ph.D. Thesis, Radboud University]. Chapter 3."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/9\/8247\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:29:19Z","timestamp":1760124559000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/9\/8247"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,5,4]]},"references-count":28,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2023,5]]}},"alternative-id":["ijms24098247"],"URL":"https:\/\/doi.org\/10.3390\/ijms24098247","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,5,4]]}}}