{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T22:26:31Z","timestamp":1787005591211,"version":"build-2736575974"},"posted":{"date-parts":[[2026,8,13]]},"group-title":"Systems Biology","reference-count":39,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T00:00:00Z","timestamp":1786579200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH","award":["K01AG078485"],"award-info":[{"award-number":["K01AG078485"]}]},{"name":"Oregon Health & Science University, Molecular and Medical Genetics Start-up Funds"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,8,13]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Krabbe disease is a rare autosomal recessive lysosomal disease caused by deficiency of galactocerebrosidase (GALC), leading to accumulation of galactosylceramide and formation of the toxic metabolite galactosylsphingosine (psychosine). While psychosine accumulation is well-established as a primary pathogenic mechanism, the broader metabolic consequences of GALC deficiency remain incompletely understood. In this study, we used stable isotope tracing to comprehensively characterize metabolic perturbations in a human oligodendrocellular Krabbe disease model. This approach revealed elevated\n                  <jats:italic>de novo<\/jats:italic>\n                  ceramide synthesis in\n                  <jats:italic>GALC<\/jats:italic>\n                  knock-out cells, characterized by increased incorporation of glucose-derived serine into ceramide biosynthetic pathways. This enhanced ceramide production was amenable to pharmacological intervention by tezacaftor, an inhibitor of sphingolipid \u0394\n                  <jats:sup>4<\/jats:sup>\n                  -desaturate (DEGS); tezacaftor administration also normalized psychosine levels, raising the possibility of its use as substrate reduction therapy. Additionally, we identified significant disruption of UDP-hexose metabolism, manifesting as an overabundance of truncated and hypogalactosylated glycans. These findings suggest impaired protein glycosylation as a previously unrecognized pathogenic mechanism in Krabbe disease. Our findings reveal novel metabolic dysregulation in Krabbe disease extending beyond established psychosine toxicity. The identification of enhanced\n                  <jats:italic>de novo<\/jats:italic>\n                  ceramide synthesis presents a new therapeutic target, while the discovery of galactose-deficient glycosylation defects supports galactose supplementation as a potential therapeutic intervention. These metabolic insights provide new mechanistic understanding and therapeutic opportunities for this devastating neurodegenerative disorder.\n                <\/jats:p>","DOI":"10.64898\/2026.08.12.744295","type":"posted-content","created":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T00:20:27Z","timestamp":1786666827000},"source":"Crossref","is-referenced-by-count":0,"title":["Metabolomic, lipidomic, and N-glycomic analyses of a human cell model of Krabbe disease reveal treatable deficits in glycosylation and serine-ceramide metabolism"],"prefix":"10.64898","author":[{"given":"Rodrigo","family":"Starosta","sequence":"first","affiliation":[{"name":"Oregon Health & Science University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hannah","family":"Saeger","sequence":"additional","affiliation":[{"name":"Oregon Health & Science University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johanna","family":"ten Hoeve","sequence":"additional","affiliation":[{"name":"University of California, Los Angeles"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"SJ","family":"Kim","sequence":"additional","affiliation":[{"name":"Oregon Health & Science University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johan L.K.","family":"Van Hove","sequence":"additional","affiliation":[{"name":"University of Colorado Anschutz"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuntian","family":"Jiang","sequence":"additional","affiliation":[{"name":"Washington University in St. Louis"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miao","family":"He","sequence":"additional","affiliation":[{"name":"Children\u2019s Hospital of Philadelphia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Neal","family":"Bennett","sequence":"additional","affiliation":[{"name":"Oregon Health & Science University"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026081715050800000_2026.08.12.744295v1.1","doi-asserted-by":"publisher","DOI":"10.1002\/jnr.490010106"},{"key":"2026081715050800000_2026.08.12.744295v1.2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1912108116"},{"key":"2026081715050800000_2026.08.12.744295v1.3","doi-asserted-by":"crossref","first-page":"e0277058","DOI":"10.1371\/journal.pone.0277058","article-title":"Galactosyl- and glucosylsphingosine induce lysosomal membrane permeabilization and cell death in cancer cells","volume":"17","year":"2022","journal-title":"PLOS ONE"},{"key":"2026081715050800000_2026.08.12.744295v1.4","doi-asserted-by":"crossref","first-page":"1260","DOI":"10.1038\/s42003-025-08684-7","article-title":"Glucosylsphingosine affects mitochondrial function in a neuronal cell model","volume":"8","year":"2025","journal-title":"Commun. Biol"},{"key":"2026081715050800000_2026.08.12.744295v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.neulet.2003.12.126"},{"key":"2026081715050800000_2026.08.12.744295v1.6","doi-asserted-by":"publisher","DOI":"10.1194\/jlr.M600084-JLR200"},{"key":"2026081715050800000_2026.08.12.744295v1.7","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1016\/j.ymgme.2013.09.011","article-title":"Characterization and application of a disease-cell model for a neurodegenerative lysosomal disease","volume":"111","year":"2014","journal-title":"Mol. Genet. Metab"},{"key":"2026081715050800000_2026.08.12.744295v1.8","doi-asserted-by":"crossref","first-page":"1361","DOI":"10.1093\/hmg\/ddac299","article-title":"Upregulation of non-canonical and canonical inflammasome genes associates with pathological features in Krabbe disease and related disorders","volume":"32","year":"2023","journal-title":"Hum. Mol. Genet"},{"key":"2026081715050800000_2026.08.12.744295v1.9","doi-asserted-by":"crossref","first-page":"e70212","DOI":"10.1002\/jimd.70212","article-title":"Validating a Human Cell Model of Null Galactosylceramidase (GALC) Enzyme Activity That Recapitulates Krabbe Disease","volume":"49","year":"2026","journal-title":"J. Inherit. Metab. Dis"},{"key":"2026081715050800000_2026.08.12.744295v1.10","doi-asserted-by":"crossref","first-page":"580","DOI":"10.3390\/biomedicines11020580","article-title":"N-Glycoprofiling of SLC35A2-CDG: Patient with a Novel Hemizygous Variant","volume":"11","year":"2023","journal-title":"Biomedicines"},{"key":"2026081715050800000_2026.08.12.744295v1.11","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1186\/s13023-021-01941-y","article-title":"Clinical, molecular and glycophenotype insights in SLC39A8-CDG","volume":"16","year":"2021","journal-title":"Orphanet J. Rare Dis"},{"key":"2026081715050800000_2026.08.12.744295v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajhg.2019.03.003"},{"key":"2026081715050800000_2026.08.12.744295v1.13","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1007\/s00018-025-05759-w","article-title":"Glycosphingolipid synthesis is impaired in SLC35A2- CDG and improves with galactose supplementation","volume":"82","year":"2025","journal-title":"Cell. Mol. Life Sci. CMLS"},{"key":"2026081715050800000_2026.08.12.744295v1.14","doi-asserted-by":"crossref","first-page":"1167","DOI":"10.1016\/j.jcf.2024.05.004","article-title":"Tezacaftor is a direct inhibitor of sphingolipid delta-4 desaturase enzyme (DEGS)","volume":"23","year":"2024","journal-title":"J. Cyst. Fibros. Off. J. Eur. Cyst. Fibros. Soc"},{"key":"2026081715050800000_2026.08.12.744295v1.15","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.0000000000004418"},{"key":"2026081715050800000_2026.08.12.744295v1.16","doi-asserted-by":"publisher","DOI":"10.1182\/blood.2020005477"},{"key":"2026081715050800000_2026.08.12.744295v1.17","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1016\/j.braindev.2023.04.001","article-title":"Favorable outcome of hematopoietic stem cell transplantation in late-onset Krabbe disease","volume":"45","year":"2023","journal-title":"Brain Dev"},{"key":"2026081715050800000_2026.08.12.744295v1.18","doi-asserted-by":"publisher","DOI":"10.1097\/GIM.0b013e3181a16e04"},{"key":"2026081715050800000_2026.08.12.744295v1.19","doi-asserted-by":"crossref","first-page":"2947","DOI":"10.1182\/bloodadvances.2021006094","article-title":"Benefits of newborn screening and hematopoietic cell transplant in infantile Krabbe disease","volume":"6","year":"2022","journal-title":"Blood Adv"},{"key":"2026081715050800000_2026.08.12.744295v1.20","doi-asserted-by":"publisher","DOI":"10.1038\/gim.2016.35"},{"key":"2026081715050800000_2026.08.12.744295v1.21","doi-asserted-by":"crossref","first-page":"888231","DOI":"10.3389\/fnmol.2022.888231","article-title":"Improved Brain Pathology and Progressive Peripheral Neuropathy in a 15 Year Old Survivor of Infantile Krabbe Disease Treated With Umbilical Cord Transplantation","volume":"15","year":"2022","journal-title":"Front. Mol. Neurosci"},{"key":"2026081715050800000_2026.08.12.744295v1.22","doi-asserted-by":"crossref","first-page":"2233","DOI":"10.1016\/j.bbmt.2018.06.020","article-title":"Long-Term Functional Outcomes after Hematopoietic Stem Cell Transplant for Early Infantile Krabbe Disease","volume":"24","year":"2018","journal-title":"Biol. Blood Marrow Transplant"},{"key":"2026081715050800000_2026.08.12.744295v1.23","doi-asserted-by":"crossref","first-page":"1094","DOI":"10.1002\/jnr.23789","article-title":"Metabolic profiling reveals biochemical pathways and potential biomarkers associated with the pathogenesis of Krabbe disease","volume":"94","year":"2016","journal-title":"J. Neurosci. Res"},{"key":"2026081715050800000_2026.08.12.744295v1.24","doi-asserted-by":"publisher","DOI":"10.1038\/jcbfm.2013.64"},{"key":"2026081715050800000_2026.08.12.744295v1.25","doi-asserted-by":"publisher","DOI":"10.1093\/brain\/awaf150"},{"key":"2026081715050800000_2026.08.12.744295v1.26","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-18084-6"},{"key":"2026081715050800000_2026.08.12.744295v1.27","doi-asserted-by":"publisher","DOI":"10.1002\/jimd.12055"},{"key":"2026081715050800000_2026.08.12.744295v1.28","doi-asserted-by":"publisher","DOI":"10.1093\/brain\/awaf198"},{"key":"2026081715050800000_2026.08.12.744295v1.29","unstructured":"Bartel, T. M. et al. Olig2-specific loss-of-function Slc35a2 results in hypomyelination and spontaneous seizures. Epilepsia n\/a,."},{"key":"2026081715050800000_2026.08.12.744295v1.30","first-page":"1102","article-title":"Clinical and biochemical improvement with galactose supplementation in SLC35A2-CDG","volume":"22","year":"2020","journal-title":"Genet. Med. Off. J. Am. Coll. Med. Genet"},{"key":"2026081715050800000_2026.08.12.744295v1.31","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgme.2014.06.002"},{"key":"2026081715050800000_2026.08.12.744295v1.32","doi-asserted-by":"crossref","first-page":"950","DOI":"10.1111\/epi.18697","article-title":"Olig2-specific loss-of-function Slc35a2 results in hypomyelination and spontaneous seizures","volume":"67","year":"2026","journal-title":"Epilepsia"},{"key":"2026081715050800000_2026.08.12.744295v1.33","doi-asserted-by":"crossref","first-page":"1261","DOI":"10.1002\/jnr.23791","article-title":"Substrate reduction therapy for Krabbe\u2019s disease","volume":"94","year":"2016","journal-title":"J. Neurosci. Res"},{"key":"2026081715050800000_2026.08.12.744295v1.34","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-4547(20000415)60:2<231::AID-JNR12>3.0.CO;2-E"},{"key":"2026081715050800000_2026.08.12.744295v1.35","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.4199-14.2015"},{"key":"2026081715050800000_2026.08.12.744295v1.36","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-021-93601-1"},{"key":"2026081715050800000_2026.08.12.744295v1.37","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2018.296780"},{"key":"2026081715050800000_2026.08.12.744295v1.38","doi-asserted-by":"publisher","DOI":"10.1139\/o59-099"},{"key":"2026081715050800000_2026.08.12.744295v1.39","doi-asserted-by":"crossref","first-page":"e4235","DOI":"10.1002\/bmc.4235","article-title":"A HILIC-MS\/MS Method for Simultaneous Quantification of the Lysosomal Disease Markers Galactosylsphingosine and Glucosylsphingosine in Mouse Serum","volume":"32","year":"2018","journal-title":"Biomed. Chromatogr. BMC"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.08.12.744295","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T22:05:17Z","timestamp":1787004317000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.08.12.744295"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,13]]},"references-count":39,"URL":"https:\/\/doi.org\/10.64898\/2026.08.12.744295","relation":{},"subject":[],"published":{"date-parts":[[2026,8,13]]},"subtype":"preprint"}}