{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T17:37:26Z","timestamp":1779385046850,"version":"3.53.1"},"reference-count":35,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2024,9,4]],"date-time":"2024-09-04T00:00:00Z","timestamp":1725408000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001711","name":"Schweizerischer Nationalfonds zur F\u00f6rderung der Wissenschaftlichen Forschung","doi-asserted-by":"publisher","award":["192691"],"award-info":[{"award-number":["192691"]}],"id":[{"id":"10.13039\/501100001711","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J of Inher Metab Disea"],"published-print":{"date-parts":[[2025,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Introduction<\/jats:title><jats:p>Metabolomic discrimination of different mitochondrial defects is challenging. We describe an NMR\u2010based bioreactor allowing real\u2010time intra\u2010 and extracellular metabolic investigation of perfused fibroblasts.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objectives<\/jats:title><jats:p>The objective of this study is (I) determining whether metabolic investigations of perfused fibroblasts overall and separated for intra\u2010 and extracellular contributions by real\u2010time NMR allows for discrimination of different representative mitochondrial defects in a feasibility study and (II) gaining insight into physiological consequences of mitochondrial dysfunction in basal condition and during glycolysis inhibition.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Overall, intra\u2010 and extracellular metabolomes of malate dehydrogenase 2 (MDH2), pyruvate dehydrogenase (PDH), complex I (CI) deficient fibroblasts, and control fibroblasts were investigated under standard culture conditions and under glycolysis inhibition. In addition to \u201coverall\u201d metabolite quantification, intra\u2010 and extracellular metabolic contributions were separated based on diffusion rate differences.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results and Discussion<\/jats:title><jats:p>Overall metabolites: Chemometric analysis of the entire metabolome revealed good separation between control, PDH and MDH2, while CI was less well separated. However, mixed intra\u2010 and extracellular changes complicated interpretation of the cellular metabolism. Intra\u2010 and extracellular metabolites: Compartment specific chemometrics revealed possibly augmenting metabolomic separation between control and deficient cell lines under basal and inhibition condition. All mitochondrial defects exhibited upregulation of glycolytic metabolism compared to controls. Inhibition of glycolysis resulted in perturbations of other metabolic pathways such as glutaminolysis, alanine, arginine, glutamate, and proline metabolism. MDH2 showed upregulation of alanine and glutamate metabolism, while the CI defect revealed lower intracellular arginine and downregulation of glutamate and arginine\u2010dependent proline synthesis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Discrimination of intra\u2010 and extracellular metabolic contributions helps understanding the underlying mechanisms of mitochondrial disorders, uncovers potential metabolic biomarkers, and unravels metabolic pathway\u2010specific adaptations in response to metabolic perturbations.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jimd.12794","type":"journal-article","created":{"date-parts":[[2024,9,5]],"date-time":"2024-09-05T02:04:30Z","timestamp":1725501870000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Intra\u2010 and extracellular real\u2010time analysis of perfused fibroblasts using an <scp>NMR<\/scp> bioreactor: A pilot study"],"prefix":"10.1002","volume":"48","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3048-9877","authenticated-orcid":false,"given":"Christian","family":"Urz\u00ec","sequence":"first","affiliation":[{"name":"Magnetic Resonance Methodology, Institute of Diagnostic and Interventional Neuroradiology University of Bern  Bern Switzerland"},{"name":"University Institute of Clinical Chemistry Inselspital, Bern University Hospital  Bern Switzerland"},{"name":"Graduate School for Cellular and Biomedical Sciences University of Bern  Bern Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-6010-300X","authenticated-orcid":false,"given":"Christoph","family":"Meyer","sequence":"additional","affiliation":[{"name":"Magnetic Resonance Methodology, Institute of Diagnostic and Interventional Neuroradiology University of Bern  Bern Switzerland"},{"name":"University Institute of Clinical Chemistry Inselspital, Bern University Hospital  Bern Switzerland"},{"name":"Graduate School for Cellular and Biomedical Sciences University of Bern  Bern Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2326-7782","authenticated-orcid":false,"given":"D\u00e9borah","family":"Mathis","sequence":"additional","affiliation":[{"name":"University Institute of Clinical Chemistry Inselspital, Bern University Hospital  Bern Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8649-1098","authenticated-orcid":false,"given":"Peter","family":"Vermathen","sequence":"additional","affiliation":[{"name":"Magnetic Resonance Methodology, Institute of Diagnostic and Interventional Neuroradiology University of Bern  Bern Switzerland"},{"name":"Translational Imaging Center (TIC) Swiss Institute for Translational and Entrepreneurial Medicine  Bern Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3650-6153","authenticated-orcid":false,"given":"Jean\u2010Marc","family":"Nuoffer","sequence":"additional","affiliation":[{"name":"University Institute of Clinical Chemistry Inselspital, Bern University Hospital  Bern Switzerland"},{"name":"Department of Pediatric Endocrinology, Diabetology and Metabolism University Children's Hospital of Bern  Bern Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,9,4]]},"reference":[{"key":"e_1_2_14_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrdp.2016.80"},{"key":"e_1_2_14_3_1","doi-asserted-by":"publisher","DOI":"10.1055\/s-0037-1603776"},{"key":"e_1_2_14_4_1","doi-asserted-by":"publisher","DOI":"10.3390\/metabo12050398"},{"key":"e_1_2_14_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/jimd.12696"},{"key":"e_1_2_14_6_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11306-019-1497-2"},{"key":"e_1_2_14_7_1","doi-asserted-by":"publisher","DOI":"10.1039\/D1AN00041A"},{"key":"e_1_2_14_8_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms23126555"},{"key":"e_1_2_14_9_1","doi-asserted-by":"publisher","DOI":"10.1042\/BST0340217"},{"key":"e_1_2_14_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/jimd.12402"},{"key":"e_1_2_14_11_1","doi-asserted-by":"publisher","DOI":"10.1136\/jmedgenet-2012-101159"},{"key":"e_1_2_14_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgmr.2021.100814"},{"key":"e_1_2_14_13_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21010234"},{"key":"e_1_2_14_14_1","doi-asserted-by":"publisher","DOI":"10.1039\/C1CC16699A"},{"key":"e_1_2_14_15_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0128478"},{"key":"e_1_2_14_16_1","doi-asserted-by":"publisher","DOI":"10.3390\/molecules26226766"},{"key":"e_1_2_14_17_1","doi-asserted-by":"publisher","DOI":"10.3390\/metabo7040061"},{"key":"e_1_2_14_18_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkl923"},{"key":"e_1_2_14_19_1","volume-title":"The Cell: A Molecular Approach","author":"Cooper GM","year":"2000"},{"key":"e_1_2_14_20_1","doi-asserted-by":"publisher","DOI":"10.1021\/ac981422i"},{"key":"e_1_2_14_21_1","doi-asserted-by":"publisher","DOI":"10.1155\/2011\/790132"},{"key":"e_1_2_14_22_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11306-018-1345-9"},{"key":"e_1_2_14_23_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.3c02314"},{"key":"e_1_2_14_24_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.8.3228"},{"key":"e_1_2_14_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/0167-4889(83)90071-X"},{"key":"e_1_2_14_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mito.2004.07.005"},{"key":"e_1_2_14_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2014.09.025"},{"key":"e_1_2_14_28_1","doi-asserted-by":"publisher","DOI":"10.1042\/EBC20170111"},{"key":"e_1_2_14_29_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-1004(200002)15:2<123::AID-HUMU1>3.0.CO;2-P"},{"key":"e_1_2_14_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajhg.2016.11.014"},{"key":"e_1_2_14_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2016.09.031"},{"key":"e_1_2_14_32_1","doi-asserted-by":"publisher","DOI":"10.1002\/jimd.12220"},{"key":"e_1_2_14_33_1","doi-asserted-by":"publisher","DOI":"10.3892\/or.2012.2085"},{"key":"e_1_2_14_34_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10863-015-9604-1"},{"key":"e_1_2_14_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymben.2013.08.005"},{"key":"e_1_2_14_36_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgme.2006.04.015"}],"container-title":["Journal of Inherited Metabolic Disease"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jimd.12794","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,8,11]],"date-time":"2025-08-11T10:08:17Z","timestamp":1754906897000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jimd.12794"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,9,4]]},"references-count":35,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2025,1]]}},"alternative-id":["10.1002\/jimd.12794"],"URL":"https:\/\/doi.org\/10.1002\/jimd.12794","archive":["Portico"],"relation":{},"ISSN":["0141-8955","1573-2665"],"issn-type":[{"value":"0141-8955","type":"print"},{"value":"1573-2665","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,9,4]]},"assertion":[{"value":"2024-02-14","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-08-19","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-09-04","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e12794"}}