{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:09:34Z","timestamp":1792804174858,"version":"4.5.0"},"reference-count":33,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T00:00:00Z","timestamp":1747699200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"KAKENHI","doi-asserted-by":"publisher","award":["23H02575"],"award-info":[{"award-number":["23H02575"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"KAKENHI","doi-asserted-by":"publisher","award":["19H03544"],"award-info":[{"award-number":["19H03544"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"KAKENHI","doi-asserted-by":"publisher","award":["23H04227"],"award-info":[{"award-number":["23H04227"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"KAKENHI","doi-asserted-by":"publisher","award":["21H05686"],"award-info":[{"award-number":["21H05686"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Grant-in-Aid for Transformative Research Areas (A)","award":["23H02575"],"award-info":[{"award-number":["23H02575"]}]},{"name":"Grant-in-Aid for Transformative Research Areas (A)","award":["19H03544"],"award-info":[{"award-number":["19H03544"]}]},{"name":"Grant-in-Aid for Transformative Research Areas (A)","award":["23H04227"],"award-info":[{"award-number":["23H04227"]}]},{"name":"Grant-in-Aid for Transformative Research Areas (A)","award":["21H05686"],"award-info":[{"award-number":["21H05686"]}]},{"name":"SENSHIN Medical Research Foundation","award":["23H02575"],"award-info":[{"award-number":["23H02575"]}],"id":[{"id":"https:\/\/ror.org\/021tj4g88","id-type":"ROR","asserted-by":"crossref"}]},{"name":"SENSHIN Medical Research Foundation","award":["19H03544"],"award-info":[{"award-number":["19H03544"]}],"id":[{"id":"https:\/\/ror.org\/021tj4g88","id-type":"ROR","asserted-by":"crossref"}]},{"name":"SENSHIN Medical Research Foundation","award":["23H04227"],"award-info":[{"award-number":["23H04227"]}],"id":[{"id":"https:\/\/ror.org\/021tj4g88","id-type":"ROR","asserted-by":"crossref"}]},{"name":"SENSHIN Medical Research Foundation","award":["21H05686"],"award-info":[{"award-number":["21H05686"]}],"id":[{"id":"https:\/\/ror.org\/021tj4g88","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Microcephaly with pontine and cerebellar hypoplasia (MICPCH) syndrome is a severe neurodevelopmental disorder caused by a deficiency in the X-linked gene calcium\/calmodulin-dependent serine protein kinase (CASK). A better understanding of the role of CASK in the pathophysiology of neurodevelopmental disorders may provide insights into novel therapeutic and diagnostic strategies for MICPCH syndrome and other neurodegenerative diseases. To investigate this, we generated CASK knockout (KO) cerebellar granule (CG) cell culture from CASK floxed (CASKflox\/flox) mice by infecting lentiviruses expressing codon-improved Cre recombinase (iCre). We performed RNA-sequencing (RNA-seq) on these cells and found that CASK-KO CG cells underwent apoptosis by activating intracellular Jun N-terminal kinase (JNK) signaling and upregulating reactive oxygen species (ROS)-related gene expression. We also performed mouse gait analysis and limb clasping behavior experiments on trans-heterozygous CASK-KO and Hprt-eGFP (CASK+\/- HprteGFP\/+) mice. The CASK+\/- HprteGFP\/+ mice exhibited cerebellar ataxic phenotypes as judged by the scores of these experiments compared to the CASK wild-type control (CASK+\/+ HprteGFP\/+) mice. Interestingly, the administration of the JNK inhibitor, JNK-IN-8, in CASK-KO CG cell cultures increased CG cell survival by reducing ROS generation. Moreover, injection of JNK-IN-8 into the cerebellum of CASK+\/- HprteGFP\/+ mice suppressed CG cell death and alleviated cerebellar ataxic phenotypes in vivo. In conclusion, JNK-IN-8 suppresses the cell death and activation of the ROS pathway in CASK-KO CG cells in both in vitro and in vivo models, suggesting its potential as a therapeutic strategy for cerebellar neurodegeneration in MICPCH syndrome.<\/jats:p>","DOI":"10.3390\/cells14100750","type":"journal-article","created":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T11:50:31Z","timestamp":1747741831000},"page":"750","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Jun N-Terminal Kinase Inhibitor Suppresses CASK Deficiency-Induced Cerebellar Granular Cell Death in MICPCH Syndrome Model Mice"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1214-359X","authenticated-orcid":false,"given":"Qi","family":"Guo","sequence":"first","affiliation":[{"name":"Department of Molecular and Cellular Physiology, Shinshu University School of Medicine, Matsumoto 390-8621, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emi","family":"Kouyama-Suzuki","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Physiology, Shinshu University School of Medicine, Matsumoto 390-8621, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3263-4986","authenticated-orcid":false,"given":"Yoshinori","family":"Shirai","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Physiology, Shinshu University School of Medicine, Matsumoto 390-8621, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1673-4637","authenticated-orcid":false,"given":"Katsuhiko","family":"Tabuchi","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Physiology, Shinshu University School of Medicine, Matsumoto 390-8621, Japan"},{"name":"Department of NeuroHealth Innovation, Institute for Biomedical Sciences, Interdisciplinary Cluster for Cutting Edge Research, Shinshu University, Matsumoto 390-8621, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1016\/j.braindev.2020.11.007","article-title":"The eldest case of MICPCH with CASK mutation exhibiting gross motor regression","volume":"43","author":"Nishio","year":"2021","journal-title":"Brain Dev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1016\/j.ajhg.2008.12.018","article-title":"A missense mutation in CASK causes FG syndrome in an Italian family","volume":"84","author":"Piluso","year":"2009","journal-title":"Am. J. Hum. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"741","DOI":"10.1136\/jmedgenet-2011-100218","article-title":"Phenotypic spectrum associated with CASK loss-of-function mutations","volume":"48","author":"Moog","year":"2011","journal-title":"J. Med. Genet."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1002\/ana.21755","article-title":"Calcium\/calmodulin-dependent serine protein kinase and mental retardation","volume":"66","author":"Hsueh","year":"2009","journal-title":"Ann. Neurol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2488","DOI":"10.1523\/JNEUROSCI.16-08-02488.1996","article-title":"CASK: A novel dlg\/PSD95 homolog with an N-terminal calmodulin-dependent protein kinase domain identified by interaction with neurexins","volume":"16","author":"Hata","year":"1996","journal-title":"J. Neurosci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1016\/j.ceb.2006.02.010","article-title":"Synaptic transmission regulated by a presynaptic MALS\/Liprin-alpha protein complex","volume":"18","author":"Olsen","year":"2006","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2525","DOI":"10.1073\/pnas.0611003104","article-title":"Deletion of CASK in mice is lethal and impairs synaptic function","volume":"104","author":"Atasoy","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1079","DOI":"10.1038\/s41380-018-0338-4","article-title":"Deficiency of calcium\/calmodulin-dependent serine protein kinase disrupts the excitatory-inhibitory balance of synapses by down-regulating GluN2B","volume":"24","author":"Mori","year":"2019","journal-title":"Mol. Psychiatry"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Guo, Q., Kouyama-Suzuki, E., Shirai, Y., Cao, X., Yanagawa, T., Mori, T., and Tabuchi, K. (2023). Structural Analysis Implicates CASK-Liprin-alpha2 Interaction in Cerebellar Granular Cell Death in MICPCH Syndrome. Cells, 12.","DOI":"10.3390\/cells12081177"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Ding, B., Bao, C., Jin, L., Xu, L., Fan, W., and Lou, W. (2021). CASK Silence Overcomes Sorafenib Resistance of Hepatocellular Carcinoma Through Activating Apoptosis and Autophagic Cell Death. Front. Oncol., 11.","DOI":"10.3389\/fonc.2021.681683"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"928","DOI":"10.1016\/j.freeradbiomed.2005.10.056","article-title":"JNK signaling pathway is a key modulator in cell death mediated by reactive oxygen and nitrogen species","volume":"40","author":"Shen","year":"2006","journal-title":"Free Radic. Biol. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"730","DOI":"10.1038\/sj.cdd.4401830","article-title":"Reactive oxygen species mediate crosstalk between NF-kappaB and JNK","volume":"13","author":"Nakano","year":"2006","journal-title":"Cell Death Differ."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/j.neuron.2013.10.051","article-title":"Cellular resolution maps of X chromosome inactivation: Implications for neural development, function, and disease","volume":"81","author":"Wu","year":"2014","journal-title":"Neuron"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"110624","DOI":"10.1016\/j.celrep.2022.110624","article-title":"Neurexins play a crucial role in cerebellar granule cell survival by organizing autocrine machinery for neurotrophins","volume":"39","author":"Uemura","year":"2022","journal-title":"Cell Rep."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Liu, Y., Zhao, Y., Guo, Q., Wang, P., Li, P., Du, Q., Xu, H., Yu, Q., Zhao, X., and Zhang, W. (2024). Sophoricoside reduces inflammation in type II collagen-induced arthritis by downregulating NLRP3 signaling. Biochem. Biophys. Rep., 40.","DOI":"10.1016\/j.bbrep.2024.101867"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1523","DOI":"10.1038\/s41467-019-09234-6","article-title":"Metascape provides a biologist-oriented resource for the analysis of systems-level datasets","volume":"10","author":"Zhou","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"15545","DOI":"10.1073\/pnas.0506580102","article-title":"Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles","volume":"102","author":"Subramanian","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e6","DOI":"10.1161\/HYPERTENSIONAHA.124.23666","article-title":"Adrenomedullin 2\/Intermedin Exerts Cardioprotective Effects by Regulating Cardiomyocyte Mitochondrial Function","volume":"82","author":"Zhao","year":"2025","journal-title":"Hypertension"},{"key":"ref_19","first-page":"e55523","article-title":"Assessment of Spontaneous Alternation, Novel Object Recognition and Limb Clasping in Transgenic Mouse Models of Amyloid-beta and Tau Neuropathology","volume":"123","author":"Miedel","year":"2017","journal-title":"J. Vis. Exp."},{"key":"ref_20","first-page":"e59878","article-title":"Low-Cost Gait Analysis for Behavioral Phenotyping of Mouse Models of Neuromuscular Disease","volume":"149","author":"Wertman","year":"2019","journal-title":"J. Vis. Exp."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2958","DOI":"10.1242\/dev.136739","article-title":"Live imaging of X chromosome reactivation dynamics in early mouse development can discriminate naive from primed pluripotent stem cells","volume":"143","author":"Kobayashi","year":"2016","journal-title":"Development"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1038\/ng.367","article-title":"A systematic, large-scale resequencing screen of X-chromosome coding exons in mental retardation","volume":"41","author":"Tarpey","year":"2009","journal-title":"Nat. Genet."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/S0896-6273(04)00139-4","article-title":"Transcriptional modification by a CASK-interacting nucleosome assembly protein","volume":"42","author":"Wang","year":"2004","journal-title":"Neuron"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/j.cell.2014.12.019","article-title":"Glial lipid droplets and ROS induced by mitochondrial defects promote neurodegeneration","volume":"160","author":"Liu","year":"2015","journal-title":"Cell"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1875","DOI":"10.2174\/138161207780858384","article-title":"JNK signalling: A possible target to prevent neurodegeneration","volume":"13","author":"Borsello","year":"2007","journal-title":"Curr. Pharm. Des."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1044","DOI":"10.1136\/jmedgenet-2021-108115","article-title":"Complete loss of the X-linked gene CASK causes severe cerebellar degeneration","volume":"59","author":"Patel","year":"2022","journal-title":"J. Med. Genet."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Tibbe, D., Pan, Y.E., Reissner, C., Harms, F.L., and Kreienkamp, H.J. (2021). Functional analysis of CASK transcript variants expressed in human brain. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0253223"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"S127","DOI":"10.3233\/JAD-201053","article-title":"c-Jun N-Terminal Kinases in Alzheimer\u2019s Disease: A Possible Target for the Modulation of the Earliest Alterations","volume":"82","author":"Busquets","year":"2021","journal-title":"J. Alzheimers Dis."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"105090","DOI":"10.1016\/j.phrs.2020.105090","article-title":"Map kinase signaling as therapeutic target for neurodegeneration","volume":"160","author":"Ahmed","year":"2020","journal-title":"Pharmacol. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1093\/jb\/mvad068","article-title":"Phosphorylated SARM1 is involved in the pathological process of rotenone-induced neurodegeneration","volume":"174","author":"Murata","year":"2023","journal-title":"J. Biochem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"417","DOI":"10.2174\/1871527320666210126113848","article-title":"Understanding Abnormal c-JNK\/p38MAPK Signaling in Amyotrophic Lateral Sclerosis: Potential Drug Targets and Influences on Neurological Disorders","volume":"20","author":"Yadav","year":"2021","journal-title":"CNS Neurol. Disord. Drug Targets"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1038\/s41421-020-00216-3","article-title":"CASK modulates the assembly and function of the Mint1\/Munc18-1 complex to regulate insulin secretion","volume":"6","author":"Zhang","year":"2020","journal-title":"Cell Discov."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1186\/s40478-016-0295-6","article-title":"X-linked intellectual disability gene CASK regulates postnatal brain growth in a non-cell autonomous manner","volume":"4","author":"Srivastava","year":"2016","journal-title":"Acta Neuropathol. Commun."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/10\/750\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:36:02Z","timestamp":1760031362000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/10\/750"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,20]]},"references-count":33,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2025,5]]}},"alternative-id":["cells14100750"],"URL":"https:\/\/doi.org\/10.3390\/cells14100750","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,20]]}}}