{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,30]],"date-time":"2026-08-30T02:05:55Z","timestamp":1788055555683,"version":"build-2784847793"},"reference-count":38,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2022,5,3]],"date-time":"2022-05-03T00:00:00Z","timestamp":1651536000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development"},{"name":"National Human Genome Research Institute"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Complex asparagine-linked glycosylation plays key roles in cellular functions, including cellular signaling, protein stability, and immune response. Previously, we characterized the appearance of a complex asparagine-linked glycosylated form of lysosome-associated membrane protein 1 (LAMP1) in the cerebellum of Npc1\u2212\/\u2212 mice. This LAMP1 form was found on activated microglia, and its appearance correlated both spatially and temporally with cerebellar Purkinje neuron loss. To test the importance of complex asparagine-linked glycosylation in NPC1 pathology, we generated NPC1 knock-out mice deficient in MGAT5, a key Golgi-resident glycosyl transferase involved in complex asparagine-linked glycosylation. Our results show that Mgat5\u2212\/\u2212:Npc1\u2212\/\u2212 mice were smaller than Mgat5+\/+:Npc1\u2212\/\u2212 mice, and exhibited earlier NPC1 disease onset and reduced lifespan. Western blot and lectin binding analyses of cerebellar extracts confirmed the reduction in complex asparagine-linked glycosylation, and the absence of the hyper-glycosylated LAMP1 previously observed. Western blot analysis of cerebellar extracts demonstrated reduced calbindin staining in Mgat5\u2212\/\u2212:Npc1\u2212\/\u2212 mice compared to Mgat5+\/+:Npc1\u2212\/\u2212 mutant mice, and immunofluorescent staining of cerebellar sections indicated decreased levels of Purkinje neurons and increased astrogliosis in Mgat5\u2212\/\u2212:Npc1\u2212\/\u2212 mice. Our results suggest that reduced asparagine-linked glycosylation increases NPC1 disease severity in mice, and leads to the hypothesis that mutations in genes involved in asparagine-linked glycosylation may contribute to disease severity progression in individuals with NPC1. To examine this with respect to MGAT5, we analyzed 111 NPC1 patients for two MGAT5 SNPs associated with multiple sclerosis; however, we did not identify an association with NPC1 phenotypic severity.<\/jats:p>","DOI":"10.3390\/ijms23095082","type":"journal-article","created":{"date-parts":[[2022,5,3]],"date-time":"2022-05-03T08:26:35Z","timestamp":1651566395000},"page":"5082","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Complex N-Linked Glycosylation: A Potential Modifier of Niemann\u2013Pick Disease, Type C1 Pathology"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2785-5941","authenticated-orcid":false,"given":"Niamh X.","family":"Cawley","sequence":"first","affiliation":[{"name":"Division of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna T.","family":"Lyons","sequence":"additional","affiliation":[{"name":"Division of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Abebe","sequence":"additional","affiliation":[{"name":"Research Animal Management Branch, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Department of Health and Human Services, Bethesda, MD 20892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachel","family":"Luke","sequence":"additional","affiliation":[{"name":"Division of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Yerger","sequence":"additional","affiliation":[{"name":"Division of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebecca","family":"Telese","sequence":"additional","affiliation":[{"name":"Division of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2524-1420","authenticated-orcid":false,"given":"Christopher A.","family":"Wassif","sequence":"additional","affiliation":[{"name":"Division of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9153-2920","authenticated-orcid":false,"given":"Joan E.","family":"Bailey-Wilson","sequence":"additional","affiliation":[{"name":"Computational and Statistical Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Department of Health and Human Services, Baltimore, MD 21224, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9397-0046","authenticated-orcid":false,"given":"Forbes D.","family":"Porter","sequence":"additional","affiliation":[{"name":"Division of Translational Medicine, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,5,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1186\/1750-1172-5-16","article-title":"Niemann-Pick disease type C","volume":"5","author":"Vanier","year":"2010","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1038\/gim.2015.25","article-title":"High incidence of unrecognized visceral\/neurological late-onset Niemann-Pick disease, type C1, predicted by analysis of massively parallel sequencing data sets","volume":"18","author":"Wassif","year":"2016","journal-title":"Genet. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"15287","DOI":"10.1073\/pnas.0807328105","article-title":"NPC2 facilitates bidirectional transfer of cholesterol between NPC1 and lipid bilayers, a step in cholesterol egress from lysosomes","volume":"105","author":"Infante","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1213","DOI":"10.1016\/j.cell.2009.03.049","article-title":"Structure of N-terminal domain of NPC1 reveals distinct subdomains for binding and transfer of cholesterol","volume":"137","author":"Kwon","year":"2009","journal-title":"Cell"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1016\/0005-2760(80)90116-2","article-title":"A lysosomal storage disorder in mice characterized by a dual deficiency of sphingomyelinase and glucocerebrosidase","volume":"619","author":"Pentchev","year":"1980","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"726","DOI":"10.1016\/j.bbamcr.2008.11.014","article-title":"Secondary lipid accumulation in lysosomal disease","volume":"1793","author":"Walkley","year":"2009","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1016\/S0301-0082(03)00114-X","article-title":"Patterned Purkinje cell death in the cerebellum","volume":"70","author":"Sarna","year":"2003","journal-title":"Prog. Neurobiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1016\/j.ymgme.2019.02.004","article-title":"Evaluation of age of death in Niemann-Pick disease, type C: Utility of disease support group websites to understand natural history","volume":"126","author":"Bianconi","year":"2019","journal-title":"Mol. Genet. Metab."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1111\/tra.12056","article-title":"Lysosomal membrane proteins and their central role in physiology","volume":"14","author":"Schwake","year":"2013","journal-title":"Traffic"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e99553","DOI":"10.15252\/embj.201899553","article-title":"Lipid-induced lysosomal damage after demyelination corrupts microglia protective function in lysosomal storage disorders","volume":"38","author":"Mitroi","year":"2019","journal-title":"EMBO J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1084\/jem.20040531","article-title":"Heat shock protein 70 promotes cell survival by inhibiting lysosomal membrane permeabilization","volume":"200","author":"Nylandsted","year":"2004","journal-title":"J. Exp. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1038\/nature08710","article-title":"Hsp70 stabilizes lysosomes and reverts Niemann-Pick disease-associated lysosomal pathology","volume":"463","author":"Kirkegaard","year":"2010","journal-title":"Nature"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1016\/j.mam.2006.08.005","article-title":"Roles of LAMP-1 and LAMP-2 in lysosome biogenesis and autophagy","volume":"27","author":"Eskelinen","year":"2006","journal-title":"Mol. Aspects Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1038\/sj.emboj.7601511","article-title":"LAMP proteins are required for fusion of lysosomes with phagosomes","volume":"26","author":"Huynh","year":"2007","journal-title":"EMBO J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1042\/BJ20050236","article-title":"NPC1 late endosomes contain elevated levels of non-esterified (\u2018free\u2019) fatty acids and an abnormally glycosylated form of the NPC2 protein","volume":"390","author":"Chen","year":"2005","journal-title":"Biochem. J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1619","DOI":"10.1111\/j.1471-4159.2010.06586.x","article-title":"Altered apolipoprotein E glycosylation is associated with Abeta(42) accumulation in an animal model of Niemann-Pick Type C disease","volume":"112","author":"Chua","year":"2010","journal-title":"J. Neurochem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2802","DOI":"10.1021\/acs.jproteome.7b00070","article-title":"Impact of the Niemann-Pick c1 Gene Mutation on the Total Cellular Glycomics of CHO Cells","volume":"16","author":"Furukawa","year":"2017","journal-title":"J. Proteome Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1074\/mcp.RA117.000129","article-title":"N-glycome of the Lysosomal Glycocalyx is Altered in Niemann-Pick Type C Disease (NPC) Model Cells","volume":"17","author":"Kosicek","year":"2018","journal-title":"Mol. Cell. Proteom."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3158","DOI":"10.4049\/jimmunol.180.5.3158","article-title":"Knockdown of Mgat5 inhibits breast cancer cell growth with activation of CD4+ T cells and macrophages","volume":"180","author":"Li","year":"2008","journal-title":"J. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1038\/73163","article-title":"Suppression of tumor growth and metastasis in Mgat5-deficient mice","volume":"6","author":"Granovsky","year":"2000","journal-title":"Nat. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1038\/35055582","article-title":"Negative regulation of T-cell activation and autoimmunity by Mgat5 N-glycosylation","volume":"409","author":"Demetriou","year":"2001","journal-title":"Nature"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Cawley, N.X., Sojka, C., Cougnoux, A., Lyons, A.T., Nicoli, E.R., Wassif, C.A., and Porter, F.D. (2020). Abnormal LAMP1 glycosylation may play a role in Niemann-Pick disease, type C pathology. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0227829"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1016\/j.jneuroim.2010.01.003","article-title":"MGAT5 alters the severity of multiple sclerosis","volume":"220","author":"Brynedal","year":"2010","journal-title":"J. Neuroimmunol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1002\/ajmg.b.30969","article-title":"Linear clinical progression, independent of age of onset, in Niemann-Pick disease, type C","volume":"153B","author":"Yanjanin","year":"2010","journal-title":"Am. J. Med. Genet. B Neuropsychiatr. Genet."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Imrie, J., Heptinstall, L., Knight, S., and Strong, K. (2015). Observational cohort study of the natural history of Niemann-Pick disease type C in the UK: A 5-year update from the UK clinical database. BMC Neurol., 15.","DOI":"10.1186\/s12883-015-0511-1"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"14876","DOI":"10.1073\/pnas.1520490112","article-title":"Glycosylation inhibition reduces cholesterol accumulation in NPC1 protein-deficient cells","volume":"112","author":"Li","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Chung, C., Elrick, M.J., Dell\u2019Orco, J.M., Qin, Z.S., Kalyana-Sundaram, S., Chinnaiyan, A.M., Shakkottai, V.G., and Lieberman, A.P. (2016). Heat Shock Protein Beta-1 Modifies Anterior to Posterior Purkinje Cell Vulnerability in a Mouse Model of Niemann-Pick Type C Disease. PLoS Genet., 12.","DOI":"10.1371\/journal.pgen.1006042"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1938","DOI":"10.1074\/mcp.M116.064949","article-title":"Quantitative Analysis of the Proteome Response to the Histone Deacetylase Inhibitor (HDACi) Vorinostat in Niemann-Pick Type C1 disease","volume":"16","author":"Subramanian","year":"2017","journal-title":"Mol. Cell. Proteom."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"E4651","DOI":"10.1073\/pnas.1720409115","article-title":"Metabolic control of T cell immune response through glycans in inflammatory bowel disease","volume":"115","author":"Dias","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"e00166","DOI":"10.14309\/ctg.0000000000000166","article-title":"Genetic Variants of the MGAT5 Gene Are Functionally Implicated in the Modulation of T Cells Glycosylation and Plasma IgG Glycome Composition in Ulcerative Colitis","volume":"11","author":"Pereira","year":"2020","journal-title":"Clin. Transl. Gastroenterol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"870","DOI":"10.1007\/s10620-018-4914-x","article-title":"Gastrointestinal Tract Pathology in a BALB\/c Niemann-Pick Disease Type C1 Null Mouse Model","volume":"63","author":"Cougnoux","year":"2018","journal-title":"Dig. Dis. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1177\/003693300505000213","article-title":"Niemann-pick disease type C and Crohn\u2019s disease","volume":"50","author":"Steven","year":"2005","journal-title":"Scott. Med. J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1060","DOI":"10.1136\/gutjnl-2015-310382","article-title":"Impaired antibacterial autophagy links granulomatous intestinal inflammation in Niemann-Pick disease type C1 and XIAP deficiency with NOD2 variants in Crohn\u2019s disease","volume":"66","author":"Schwerd","year":"2017","journal-title":"Gut"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1563","DOI":"10.1007\/s00432-015-1917-2","article-title":"Role of tumor cell surface lysosome-associated membrane protein-1 (LAMP1) and its associated carbohydrates in lung metastasis","volume":"141","author":"Agarwal","year":"2015","journal-title":"J. Cancer Res. Clin. Oncol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/j.jneuroim.2010.10.026","article-title":"MGAT5 and disease severity in progressive multiple sclerosis","volume":"230","author":"Esposito","year":"2011","journal-title":"J. Neuroimmunol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"797","DOI":"10.2147\/IJGM.S293675","article-title":"Novel Mutations in NPC1 are Associated with Pelizaeus-Merzbacher-Like Disease: A Case Report","volume":"14","author":"Fu","year":"2021","journal-title":"Int. J. Gen. Med."},{"key":"ref_37","first-page":"e1787","article-title":"A simple composite phenotype scoring system for evaluating mouse models of cerebellar ataxia","volume":"39","author":"Guyenet","year":"2010","journal-title":"J. Vis. Exp."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Cawley, N.X., Lyons, A.T., Abebe, D., Wassif, C.A., and Porter, F.D. (2020). Evaluation of the Potential Role of Proprotein Convertase Subtilisin\/Kexin Type 9 (PCSK9) in Niemann-Pick Disease, Type C1. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21072430"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/9\/5082\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:05:44Z","timestamp":1760137544000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/9\/5082"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,5,3]]},"references-count":38,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2022,5]]}},"alternative-id":["ijms23095082"],"URL":"https:\/\/doi.org\/10.3390\/ijms23095082","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,5,3]]}}}