{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T17:26:37Z","timestamp":1790011597660,"version":"4.0.1"},"reference-count":45,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T00:00:00Z","timestamp":1765152000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Science and Technology Council","award":["NSTC 112-2320-B-195 -001"],"award-info":[{"award-number":["NSTC 112-2320-B-195 -001"]}]},{"name":"National Science and Technology Council","award":["109-2314-B-195 -015 -MY3"],"award-info":[{"award-number":["109-2314-B-195 -015 -MY3"]}]},{"DOI":"10.13039\/501100004861","name":"MacKay Memorial Hospital","doi-asserted-by":"crossref","award":["MMH-E-114-02"],"award-info":[{"award-number":["MMH-E-114-02"]}],"id":[{"id":"10.13039\/501100004861","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100004861","name":"MacKay Memorial Hospital","doi-asserted-by":"crossref","award":["MMH-E-111-02"],"award-info":[{"award-number":["MMH-E-111-02"]}],"id":[{"id":"10.13039\/501100004861","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Globoid cell leukodystrophy (GLD) is a devastating lysosomal storage disorder caused by galactocerebrosidase (GALC) deficiency, leading to cytotoxic psychosine accumulation, broad neuroinflammation, dysfunction of autophagy and ubiquitin-proteasome system, progressive demyelination in both the central (CNS) and peripheral nervous systems (PNS), and premature death. Curative treatments are lacking, highlighting the urgent need for transformative approaches. Existing therapies have failed to achieve durable metabolic correction across neural compartments or sustained functional recovery. Here, we demonstrate that a single intracranial administration of high-titer AAV9-GALC targeting the thalamus and deep cerebellar nuclei achieves unprecedented and lifelong therapeutic efficacy in the Twitcher mouse model of GLD. This region-specific monotherapy achieved broad neuronal and glial transduction throughout the CNS and PNS, resulting in sustained supraphysiological GALC activity and complete normalization of psychosine levels. Treated mice exhibited preserved proteostasis, axonal architecture, and myelin integrity, inhibition of neuroinflammation, alongside restored motor function. Remarkably, treated mice attain lifespans approaching wild-type levels, far surpassing all previously reported interventions in this model, indicating a durable, possibly lifelong therapeutic effect. By achieving durable and comprehensive metabolic and structural correction across neural systems without repeated dosing, multi-route delivery, combinational therapy, hematopoietic stem cell transplantation, or high-dose systemic delivery, this study establishes CNS-directed AAV9 monotherapy as a clinically translatable and potentially lifelong therapeutic paradigm for GLD.<\/jats:p>","DOI":"10.3390\/cells14241942","type":"journal-article","created":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T10:35:51Z","timestamp":1765190151000},"page":"1942","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Durable Global Correction of CNS and PNS and Lifespan Rescue in Murine Globoid Cell Leukodystrophy via AAV9-Mediated Monotherapy"],"prefix":"10.3390","volume":"14","author":[{"given":"Dar-Shong","family":"Lin","sequence":"first","affiliation":[{"name":"Department of Translational Medicine, MacKay Memorial Hospital, Taipei 104217, Taiwan"},{"name":"Department of Medicine, MacKay Medical University, New Taipei 252005, Taiwan"},{"name":"Department of Pediatrics, MacKay Memorial Hospital, Taipei 104217, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7942-5823","authenticated-orcid":false,"given":"Che-Sheng","family":"Ho","sequence":"additional","affiliation":[{"name":"Department of Medicine, MacKay Medical University, New Taipei 252005, Taiwan"},{"name":"Department of Neurology, MacKay Children\u2019s Hospital, Taipei 104217, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3889-0531","authenticated-orcid":false,"given":"Yu-Wen","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Medical Research, MacKay Memorial Hospital, Taipei 104217, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tsung-Han","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Medical Research, MacKay Memorial Hospital, Taipei 104217, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zo-Darr","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Medical Research, MacKay Memorial Hospital, Taipei 104217, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tuan-Jen","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, MacKay Memorial Hospital, Taipei 104217, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5536-5942","authenticated-orcid":false,"given":"Wern-Cherng","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, MacKay Memorial Hospital, Taipei 104217, Taiwan"},{"name":"Department of Laboratory Medicine, School of Medicine, College of Medicine, National Taiwan University, Taipei 106319, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sung-Fu","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, MacKay Memorial Hospital, Taipei 104217, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1177\/08830738030180090201","article-title":"Globoid cell leukodystrophy (Krabbe\u2019s disease): Update","volume":"18","author":"Suzuki","year":"2003","journal-title":"J. Child Neurol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3064","DOI":"10.1002\/acn3.52114","article-title":"Infantile Krabbe disease (0\u201312 months), progression, and recommended endpoints for clinical trials","volume":"11","author":"Greco","year":"2024","journal-title":"Ann. Clin. Transl. Neurol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"6068","DOI":"10.1523\/JNEUROSCI.5597-08.2009","article-title":"Psychosine accumulates in membrane microdomains in the brain of krabbe patients, disrupting the raft architecture","volume":"29","author":"White","year":"2009","journal-title":"J. Neurosci."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Lin, D.S., Ho, C.S., Huang, Y.W., Wu, T.Y., Lee, T.H., Huang, Z.D., Wang, T.J., Yang, S.J., and Chiang, M.F. (2020). Impairment of Proteasome and Autophagy Underlying the Pathogenesis of Leukodystrophy. Cells, 9.","DOI":"10.3390\/cells9051124"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/j.nbd.2019.05.011","article-title":"Dysregulated autophagy as a new aspect of the molecular pathogenesis of Krabbe disease","volume":"129","author":"Angella","year":"2019","journal-title":"Neurobiol. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1002\/jemt.1070320304","article-title":"Murine model of genetic demyelinating disease: The twitcher mouse","volume":"32","author":"Suzuki","year":"1995","journal-title":"Microsc. Res. Tech."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"178","DOI":"10.1016\/0006-8993(88)91220-6","article-title":"Pathology of the peripheral nerve in the twitcher mouse following bone marrow transplantation","volume":"460","author":"Kondo","year":"1988","journal-title":"Brain Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1549","DOI":"10.1096\/fj.05-3826fje","article-title":"Enzyme replacement therapy results in substantial improvements in early clinical phenotype in a mouse model of globoid cell leukodystrophy","volume":"19","author":"Lee","year":"2005","journal-title":"FASEB J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1203\/00006450-200201000-00009","article-title":"Substrate-reduction therapy enhances the benefits of bone marrow transplantation in young mice with globoid cell leukodystrophy","volume":"51","author":"Biswas","year":"2002","journal-title":"Pediatr. Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.ymthe.2022.09.017","article-title":"Preclinical studies in Krabbe disease: A model for the investigation of novel combination therapies for lysosomal storage diseases","volume":"31","author":"Heller","year":"2023","journal-title":"Mol. Ther."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1118","DOI":"10.1046\/j.1471-4159.1996.66031118.x","article-title":"Molecular cloning and expression of cDNA for murine galactocerebrosidase and mutation analysis of the twitcher mouse, a model of Krabbe\u2019s disease","volume":"66","author":"Sakai","year":"1996","journal-title":"J. Neurochem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1373\/clinchem.2008.115873","article-title":"Specific determination of beta-galactocerebrosidase activity via competitive inhibition of beta-galactosidase","volume":"55","author":"Martino","year":"2009","journal-title":"Clin. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Matsumoto, S.I., Sato, S., Otake, K., and Kosugi, Y. (2022). Highly-sensitive simultaneous quantitation of glucosylsphingosine and galactosylsphingosine in human cerebrospinal fluid by liquid chromatography\/tandem mass spectrometry. J. Pharm. Biomed. Anal., 217.","DOI":"10.1016\/j.jpba.2022.114852"},{"key":"ref_14","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","author":"Ribbens","year":"2014","journal-title":"Mol. Genet. Metab."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1002\/jnr.20169","article-title":"Analysis of galactocerebrosidase activity in the mouse brain by a new histological staining method","volume":"77","author":"Dolcetta","year":"2004","journal-title":"J. Neurosci. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1038\/sj.mt.6300026","article-title":"Central nervous system-directed AAV2\/5-mediated gene therapy synergizes with bone marrow transplantation in the murine model of globoid-cell leukodystrophy","volume":"15","author":"Lin","year":"2007","journal-title":"Mol. Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1016\/j.ymthe.2005.04.019","article-title":"AAV2\/5 vector expressing galactocerebrosidase ameliorates CNS disease in the murine model of globoid-cell leukodystrophy more efficiently than AAV2","volume":"12","author":"Lin","year":"2005","journal-title":"Mol. Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1007\/s003359900551","article-title":"Behavioral and functional analysis of mouse phenotype: SHIRPA, a proposed protocol for comprehensive phenotype assessment","volume":"8","author":"Rogers","year":"1997","journal-title":"Mamm. Genome"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/S0165-0270(96)02211-X","article-title":"Pole test is a useful method for evaluating the mouse movement disorder caused by striatal dopamine depletion","volume":"73","author":"Matsuura","year":"1997","journal-title":"J. Neurosci. Methods"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1007\/BF00293398","article-title":"Spacio-temporal progression of demyelination in twitcher mouse: With clinico-pathological correlation","volume":"88","author":"Taniike","year":"1994","journal-title":"Acta Neuropathol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/0006-8993(93)90692-G","article-title":"Proliferation of microglia\/macrophages in the demyelinating CNS and PNS of twitcher mouse","volume":"602","author":"Ohno","year":"1993","journal-title":"Brain Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1002\/jnr.23804","article-title":"Neuroimmune mechanisms in Krabbe\u2019s disease","volume":"94","author":"Potter","year":"2016","journal-title":"J. Neurosci. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1049","DOI":"10.1002\/jnr.23773","article-title":"A microglial hypothesis of globoid cell leukodystrophy pathology","volume":"94","author":"Nicaise","year":"2016","journal-title":"J. Neurosci. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2082","DOI":"10.1093\/hmg\/ddab159","article-title":"Expression of Ripk1 and DAM genes correlates with severity and progression of Krabbe disease","volume":"30","author":"Wang","year":"2021","journal-title":"Hum. Mol. Genet."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"950","DOI":"10.1177\/0192623321991469","article-title":"Quantitative Assessment of Neuroinflammation, Myelinogenesis, Demyelination, and Nerve Fiber Regeneration in Immunostained Sciatic Nerves from Twitcher Mice with a Tissue Image Analysis Platform","volume":"49","author":"Wilson","year":"2021","journal-title":"Toxicol. Pathol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1007\/BF00687612","article-title":"Demyelination in the spinal cord of murine globoid cell leukodystrophy (the twitcher mouse)","volume":"62","author":"Takahashi","year":"1984","journal-title":"Acta Neuropathol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"AN20110019","DOI":"10.1042\/AN20110019","article-title":"Peripheral neuropathy in the Twitcher mouse involves the activation of axonal caspase 3","volume":"3","author":"Smith","year":"2011","journal-title":"ASN Neuro"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"734","DOI":"10.1016\/j.ymthe.2004.12.020","article-title":"AAV-mediated expression of galactocerebrosidase in brain results in attenuated symptoms and extended life span in murine models of globoid cell leukodystrophy","volume":"11","author":"Rafi","year":"2005","journal-title":"Mol. Ther."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1016\/j.ymgme.2011.05.005","article-title":"CNS-targeted AAV5 gene transfer results in global dispersal of vector and prevention of morphological and function deterioration in CNS of globoid cell leukodystrophy mouse model","volume":"103","author":"Lin","year":"2011","journal-title":"Mol. Genet. Metab."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.gene.2015.06.049","article-title":"Mitigation of cerebellar neuropathy in globoid cell leukodystrophy mice by AAV-mediated gene therapy","volume":"571","author":"Lin","year":"2015","journal-title":"Gene"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"9945","DOI":"10.1523\/JNEUROSCI.1802-11.2011","article-title":"Bone marrow transplantation augments the effect of brain- and spinal cord-directed adeno-associated virus 2\/5 gene therapy by altering inflammation in the murine model of globoid-cell leukodystrophy","volume":"31","author":"Reddy","year":"2011","journal-title":"J. Neurosci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2031","DOI":"10.1038\/mt.2012.153","article-title":"Extended normal life after AAVrh10-mediated gene therapy in the mouse model of Krabbe disease","volume":"20","author":"Rafi","year":"2012","journal-title":"Mol. Ther."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"135","DOI":"10.34172\/bi.2021.21","article-title":"Can early treatment of twitcher mice with high dose AAVrh10-GALC eliminate the need for BMT?","volume":"11","author":"Rafi","year":"2021","journal-title":"Bioimpacts"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"874","DOI":"10.1016\/j.ymthe.2018.01.009","article-title":"Long-Term Improvement of Neurological Signs and Metabolic Dysfunction in a Mouse Model of Krabbe\u2019s Disease after Global Gene Therapy","volume":"26","author":"Marshall","year":"2018","journal-title":"Mol. Ther."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1016\/j.ymthe.2020.12.031","article-title":"Enhanced Efficacy and Increased Long-Term Toxicity of CNS-Directed, AAV-Based Combination Therapy for Krabbe Disease","volume":"29","author":"Li","year":"2021","journal-title":"Mol. Ther."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1138","DOI":"10.1002\/jnr.23882","article-title":"Intrathecal administration of AAV\/GALC vectors in 10\u201311-day-old twitcher mice improves survival and is enhanced by bone marrow transplant","volume":"94","author":"Marshall","year":"2016","journal-title":"J. Neurosci. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1089\/hum.2013.189","article-title":"Adeno-associated virus serotypes 1, 8, and 9 share conserved mechanisms for anterograde and retrograde axonal transport","volume":"25","author":"Castle","year":"2014","journal-title":"Hum. Gene Ther."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"957","DOI":"10.1152\/physrev.1974.54.4.957","article-title":"Cerebrocerebellar communication systems","volume":"54","author":"Allen","year":"1974","journal-title":"Physiol. Rev."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2322","DOI":"10.1152\/jn.00339.2011","article-title":"The organization of the human cerebellum estimated by intrinsic functional connectivity","volume":"106","author":"Buckner","year":"2011","journal-title":"J. Neurophysiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1046\/j.1440-1681.1999.02987.x","article-title":"Barrier mechanisms in the brain, II. Immature brain","volume":"26","author":"Saunders","year":"1999","journal-title":"Clin. Exp. Pharmacol. Physiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1883","DOI":"10.1016\/j.ymthe.2021.01.026","article-title":"Waning efficacy in a long-term AAV-mediated gene therapy study in the murine model of Krabbe disease","volume":"29","author":"Heller","year":"2021","journal-title":"Mol. Ther."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1007\/s00401-011-0814-2","article-title":"Axonopathy is a compounding factor in the pathogenesis of Krabbe disease","volume":"122","author":"Castelvetri","year":"2011","journal-title":"Acta Neuropathol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"10048","DOI":"10.1523\/JNEUROSCI.0217-13.2013","article-title":"The sphingolipid psychosine inhibits fast axonal transport in Krabbe disease by activation of GSK3\u03b2 and deregulation of molecular motors","volume":"33","author":"Givogri","year":"2013","journal-title":"J. Neurosci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1016\/j.nbd.2012.01.013","article-title":"Psychosine induces the dephosphorylation of neurofilaments by deregulation of PP1 and PP2A phosphatases","volume":"46","author":"Zhu","year":"2012","journal-title":"Neurobiol. Dis."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"5356","DOI":"10.1038\/s41467-020-19179-w","article-title":"Brainstem development requires galactosylceramidase and is critical for pathogenesis in a model of Krabbe disease","volume":"11","author":"Weinstock","year":"2020","journal-title":"Nat. Commun."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/24\/1942\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,10]],"date-time":"2025-12-10T05:23:52Z","timestamp":1765344232000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/24\/1942"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,8]]},"references-count":45,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["cells14241942"],"URL":"https:\/\/doi.org\/10.3390\/cells14241942","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,8]]}}}