{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T11:45:48Z","timestamp":1791978348543,"version":"4.4.0"},"reference-count":41,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2022,4,9]],"date-time":"2022-04-09T00:00:00Z","timestamp":1649462400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"UMHS-PUHSC Joint Institute for Translational and Clinical Research","award":["BMU2019JI009"],"award-info":[{"award-number":["BMU2019JI009"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81771584, 81971390, 82071264 and 82101941"],"award-info":[{"award-number":["81771584, 81971390, 82071264 and 82101941"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Natural Science Foundation of Beijing, China","award":["7214256"],"award-info":[{"award-number":["7214256"]}]},{"name":"Beijing Key Laboratory of Molecular Diagnosis and Study on Pediatric Genetic Diseases","award":["BZ0317"],"award-info":[{"award-number":["BZ0317"]}]},{"DOI":"10.13039\/501100019018","name":"CAMS Initiative for Innovative Medicine","doi-asserted-by":"publisher","award":["2016-I2M-1-008"],"award-info":[{"award-number":["2016-I2M-1-008"]}],"id":[{"id":"10.13039\/501100019018","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Public service development and reform pilot project of Beijing Medical Research Institute","award":["BMR2019-11"],"award-info":[{"award-number":["BMR2019-11"]}]},{"name":"the Pediatric Medical Coordinated Development Center of Beijing Hospitals Authority","award":["XTZD20180402"],"award-info":[{"award-number":["XTZD20180402"]}]},{"name":"National Institute of Neurological Disorders and Stroke AREA","award":["2R15NS101594 and R03NS123728"],"award-info":[{"award-number":["2R15NS101594 and R03NS123728"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Hypomyelinating leukodystrophies (HLDs) are a rare group of heterogeneously genetic disorders characterized by persistent deficit of myelin observed on magnetic resonance imaging (MRI). To identify a new disease-associated gene of HLD, trio-based whole exome sequencing was performed for unexplained patients with HLD. Functional studies were performed to confirm the phenotypic effect of candidate protein variants. Two de novo heterozygous variants, c.227T&gt;G p.(L76R) or c.227T&gt;C p.(L76P) in TMEM163 were identified in two unrelated HLD patients. TMEM163 protein is a zinc efflux transporter localized within the plasma membrane, lysosomes, early endosomes, and other vesicular compartments. It has not been associated with hypomyelination. Functional zinc flux assays in HeLa cells stably-expressing TMEM163 protein variants, L76R and L76P, revealed distinct attenuation or enhancement of zinc efflux, respectively. Experiments using a zebrafish model with knockdown of tmem163a and tmem163b (morphants) showed that loss of tmem163 causes dysplasia of the larvae, locomotor disability and myelin deficit. Expression of human wild type TMEM163 mRNAs in morphants rescues the phenotype, while the TMEM163 L76P and L76R mutants aggravated the condition. Moreover, poor proliferation, elevated apoptosis of oligodendrocytes, and reduced oligodendrocytes and neurons were also observed in zebrafish morphants. Our findings suggest an unappreciated role for TMEM163 protein in myelin development and add TMEM163 to a growing list of genes associated with hypomyelination leukodystrophy.<\/jats:p>","DOI":"10.3390\/cells11081285","type":"journal-article","created":{"date-parts":[[2022,4,10]],"date-time":"2022-04-10T06:02:54Z","timestamp":1649570574000},"page":"1285","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Functional Study of TMEM163 Gene Variants Associated with Hypomyelination Leukodystrophy"],"prefix":"10.3390","volume":"11","author":[{"given":"Huifang","family":"Yan","sequence":"first","affiliation":[{"name":"Department of Pediatrics, Peking University First Hospital, Beijing 100034, China"},{"name":"Joint International Research Center of Translational and Clinical Research, Beijing 100191, China"},{"name":"Beijing Key Laboratory of Molecular Diagnosis and Study on Pediatric Genetic Diseases, Beijing 100034, China"},{"name":"Michigan Neuroscience Institute, University of Michigan, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1525-6415","authenticated-orcid":false,"given":"Shuyan","family":"Yang","sequence":"additional","affiliation":[{"name":"Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiming","family":"Hou","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Membrane Biology, Institute of Zoology, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Beijing 100101, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saima","family":"Ali","sequence":"additional","affiliation":[{"name":"Department of Biological Science, California State University, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrian","family":"Escobar","sequence":"additional","affiliation":[{"name":"Department of Biological Science, California State University, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Gao","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Peking University First Hospital, Beijing 100034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruoyu","family":"Duan","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Peking University First Hospital, Beijing 100034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Kubisiak","sequence":"additional","affiliation":[{"name":"Michigan Neuroscience Institute, University of Michigan, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junyu","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Peking University First Hospital, Beijing 100034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Peking University First Hospital, Beijing 100034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiangxi","family":"Xiao","sequence":"additional","affiliation":[{"name":"Department of Radiology, Peking University First Hospital, Beijing 100034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuwu","family":"Jiang","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Peking University First Hospital, Beijing 100034, China"},{"name":"Beijing Key Laboratory of Molecular Diagnosis and Study on Pediatric Genetic Diseases, Beijing 100034, China"},{"name":"Key Laboratory for Neuroscience, Ministry of Education\/National Health and Family Planning Commission, Peking University, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting","family":"Zhang","sequence":"additional","affiliation":[{"name":"Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ye","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Peking University First Hospital, Beijing 100034, China"},{"name":"Beijing Key Laboratory of Molecular Diagnosis and Study on Pediatric Genetic Diseases, Beijing 100034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Margit","family":"Burmeister","sequence":"additional","affiliation":[{"name":"Michigan Neuroscience Institute, University of Michigan, Ann Arbor, MI 48109, USA"},{"name":"Departments of Computational Medicine & Bioinformatics, Psychiatry and Human Genetics, University of Michigan, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiang","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Membrane Biology, Institute of Zoology, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Beijing 100101, China"},{"name":"Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0749-9564","authenticated-orcid":false,"given":"Math P.","family":"Cuajungco","sequence":"additional","affiliation":[{"name":"Department of Biological Science, California State University, Fullerton, CA 92831, USA"},{"name":"Center for Applied Biotechnology Studies, California State University, Fullerton, CA 92831, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingmin","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Peking University First Hospital, Beijing 100034, China"},{"name":"Joint International Research Center of Translational and Clinical Research, Beijing 100191, China"},{"name":"Beijing Key Laboratory of Molecular Diagnosis and Study on Pediatric Genetic Diseases, Beijing 100034, China"},{"name":"Key Laboratory for Neuroscience, Ministry of Education\/National Health and Family Planning Commission, Peking University, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,4,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1007\/s10545-005-5952-z","article-title":"Magnetic resonance techniques in the assessment of myelin and myelination","volume":"28","author":"Barkovich","year":"2005","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Van Der Knaap, M.S., and Valk, J. (2005). Magnetic Resonance of Myelination and Myelin Disorders, Springer. [3rd ed.].","DOI":"10.1007\/3-540-27660-2"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"903","DOI":"10.3174\/ajnr.A6051","article-title":"Oculodentodigital Dysplasia: A Hypomyelinating Leukodystrophy with a Characteristic MRI Pattern of Brain Stem Involvement","volume":"40","author":"Harting","year":"2019","journal-title":"AJNR Am. J. Neuroradiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1898","DOI":"10.1212\/WNL.0000000000001002","article-title":"Clinical spectrum of 4H leukodystrophy caused by POLR3A and POLR3B mutations","volume":"83","author":"Wolf","year":"2014","journal-title":"Neurology"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"996","DOI":"10.1016\/j.ajhg.2019.09.011","article-title":"Heterozygous Variants in the Mechanosensitive Ion Channel TMEM63A Result in Transient Hypomyelination during Infancy","volume":"105","author":"Yan","year":"2019","journal-title":"Am. J. Hum. Genet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3105","DOI":"10.1093\/brain\/awx314","article-title":"A recurrent de novo mutation in TMEM106B causes hypomyelinating leukodystrophy","volume":"140","author":"Simons","year":"2017","journal-title":"Brain"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1038\/s10038-017-0363-1","article-title":"Homozygosity for a nonsense variant in AIMP2 is associated with a progressive neurodevelopmental disorder with microcephaly, seizures, and spastic quadriparesis","volume":"63","author":"Shukla","year":"2018","journal-title":"J. Hum. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1240","DOI":"10.1172\/JCI123959","article-title":"Loss of the sphingolipid desaturase DEGS1 causes hypomyelinating leukodystrophy","volume":"129","author":"Pant","year":"2019","journal-title":"J. Clin. Investig."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1002\/ana.24194","article-title":"Hypomyelinating leukodystrophies: Translational research progress and prospects","volume":"76","author":"Pouwels","year":"2014","journal-title":"Ann. Neurol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"962","DOI":"10.1016\/S1474-4422(19)30143-7","article-title":"Diagnosis, prognosis, and treatment of leukodystrophies","volume":"18","author":"Schiffmann","year":"2019","journal-title":"Lancet Neurol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1111\/j.1471-4159.2007.04758.x","article-title":"Identification and characterization of SV31, a novel synaptic vesicle membrane protein and potential transporter","volume":"103","author":"Burre","year":"2007","journal-title":"J. Neurochem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1111\/j.1471-4159.2011.07344.x","article-title":"SV31 is a Zn2+-binding synaptic vesicle protein","volume":"118","author":"Barth","year":"2011","journal-title":"J. Neurochem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1022","DOI":"10.1016\/j.bbrc.2019.11.117","article-title":"Role of Tmem163 in zinc-regulated insulin storage of MIN6 cells: Functional exploration of an Indian type 2 diabetes GWAS associated gene","volume":"522","author":"Chakraborty","year":"2020","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.neuron.2015.11.013","article-title":"Purification and Characterization of Progenitor and Mature Human Astrocytes Reveals Transcriptional and Functional Differences with Mouse","volume":"89","author":"Zhang","year":"2016","journal-title":"Neuron"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"108166","DOI":"10.1016\/j.abb.2019.108166","article-title":"Transmembrane 163 (TMEM163) protein effluxes zinc","volume":"677","author":"Sanchez","year":"2019","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Styrpejko, D.J., and Cuajungco, M.P. (2021). Transmembrane 163 (TMEM163) Protein: A New Member of the Zinc Efflux Transporter Family. Biomedicines, 9.","DOI":"10.3390\/biomedicines9020220"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1038\/s10038-020-00896-5","article-title":"Genetic analysis of 20 patients with hypomyelinating leukodystrophy by trio-based whole-exome sequencing","volume":"66","author":"Yan","year":"2021","journal-title":"J. Hum. Genet."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"100050","DOI":"10.1016\/j.xpro.2020.100050","article-title":"Protocol for quantifying zinc flux in cultured cells using fluorescent indicators","volume":"1","author":"Ali","year":"2020","journal-title":"STAR Protoc."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"749","DOI":"10.1152\/physrev.00035.2014","article-title":"The Physiological, Biochemical, and Molecular Roles of Zinc Transporters in Zinc Homeostasis and Metabolism","volume":"95","author":"Kambe","year":"2015","journal-title":"Physiol. Rev."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Petrovski, S., Gussow, A.B., Wang, Q., Halvorsen, M., Han, Y., Weir, W.H., Allen, A.S., and Goldstein, D.B. (2015). The Intolerance of Regulatory Sequence to Genetic Variation Predicts Gene Dosage Sensitivity. PLoS Genet., 11.","DOI":"10.1371\/journal.pgen.1005492"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1038\/nature19057","article-title":"Analysis of protein-coding genetic variation in 60,706 humans","volume":"536","author":"Lek","year":"2016","journal-title":"Nature"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1247","DOI":"10.1111\/tra.12205","article-title":"Cellular zinc levels are modulated by TRPML1-TMEM163 interaction","volume":"15","author":"Cuajungco","year":"2014","journal-title":"Traffic"},{"key":"ref_23","unstructured":"The GTEx Consortium (2015). Human genomics. The Genotype-Tissue Expression (GTEx) pilot analysis: Multitissue gene regulation in humans. Science, 348, 648\u2013660."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1111\/jnc.13886","article-title":"The synaptic vesicle protein SV31 assembles into a dimer and transports Zn(2)","volume":"140","author":"Waberer","year":"2017","journal-title":"J. Neurochem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1002\/glia.22897","article-title":"Insights into mechanisms of central nervous system myelination using zebrafish","volume":"64","author":"Czopka","year":"2016","journal-title":"Glia"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"592","DOI":"10.1002\/dvdy.22166","article-title":"Visualization of myelination in GFP-transgenic zebrafish","volume":"239","author":"Jung","year":"2010","journal-title":"Dev. Dyn."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1001\/jamaneurol.2020.5257","article-title":"Identification of Candidate Parkinson Disease Genes by Integrating Genome-Wide Association Study, Expression, and Epigenetic Data Sets","volume":"78","author":"Kia","year":"2021","journal-title":"JAMA Neurol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"299.e1","DOI":"10.1016\/j.neurobiolaging.2020.11.005","article-title":"Mutation analysis of TMEM family members for early-onset Parkinson\u2019s disease in Chinese population","volume":"101","author":"Li","year":"2020","journal-title":"Neurobiol. Aging"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"977","DOI":"10.2337\/db12-0406","article-title":"Genome-wide association study for type 2 diabetes in Indians identifies a new susceptibility locus at 2q21","volume":"62","author":"Tabassum","year":"2013","journal-title":"Diabetes"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1804","DOI":"10.1182\/blood.2020007389","article-title":"A zinc transporter, transmembrane protein 163 (TMEM163), is critical for the biogenesis of platelet dense granules","volume":"137","author":"Yuan","year":"2021","journal-title":"Blood"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1007\/s00424-008-0523-4","article-title":"The varitint-waddler mouse phenotypes and the TRPML3 ion channel mutation: Cause and consequence","volume":"457","author":"Cuajungco","year":"2008","journal-title":"Pflug. Arch."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"780","DOI":"10.1038\/nrn2734","article-title":"Zinc in the physiology and pathology of the CNS","volume":"10","author":"Sensi","year":"2009","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"16076","DOI":"10.1523\/JNEUROSCI.3454-11.2011","article-title":"The neurophysiology and pathology of brain zinc","volume":"31","author":"Sensi","year":"2011","journal-title":"J. Neurosci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"750","DOI":"10.1002\/glia.22470","article-title":"Cytosolic zinc accumulation contributes to excitotoxic oligodendroglial death","volume":"61","author":"Mato","year":"2013","journal-title":"Glia"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"350","DOI":"10.2119\/2007-00042.Zhang","article-title":"Intracellular zinc release, 12-lipoxygenase activation and MAPK dependent neuronal and oligodendroglial death","volume":"13","author":"Zhang","year":"2007","journal-title":"Mol. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"828","DOI":"10.1002\/jnr.23208","article-title":"Reduced extracellular zinc levels facilitate glutamate-mediated oligodendrocyte death after trauma","volume":"91","author":"Johnstone","year":"2013","journal-title":"J. Neurosci. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"107704","DOI":"10.1016\/j.celrep.2020.107704","article-title":"TMEM163 Regulates ATP-Gated P2X Receptor and Behavior","volume":"31","author":"Salm","year":"2020","journal-title":"Cell Rep."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1874","DOI":"10.12688\/f1000research.21142.1","article-title":"Falco: High-speed FastQC emulation for quality control of sequencing data","volume":"8","author":"Smith","year":"2019","journal-title":"F1000Res"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Patel, R.K., and Jain, M. (2012). NGS QC Toolkit: A toolkit for quality control of next generation sequencing data. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0030619"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1038\/nbt.3122","article-title":"StringTie enables improved reconstruction of a transcriptome from RNA-seq reads","volume":"33","author":"Pertea","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"R106","DOI":"10.1186\/gb-2010-11-10-r106","article-title":"Differential expression analysis for sequence count data","volume":"11","author":"Anders","year":"2010","journal-title":"Genome Biol."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/8\/1285\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:51:09Z","timestamp":1760136669000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/8\/1285"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,4,9]]},"references-count":41,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2022,4]]}},"alternative-id":["cells11081285"],"URL":"https:\/\/doi.org\/10.3390\/cells11081285","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,4,9]]}}}