{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T19:29:57Z","timestamp":1789414197861,"version":"build-2803163510"},"reference-count":50,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2023,11,27]],"date-time":"2023-11-27T00:00:00Z","timestamp":1701043200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Agency (JST)"},{"name":"Grants-in-Aid for Scientific Research from the Japanese Ministry of Education, Culture, Sports, Science and Technology (MEXT)"},{"name":"Grants-in-Aid for Medical Scientific Research from the Japanese Ministry of Health, Labour and Welfare (MHLW)"},{"name":"Daiichi Sankyo Science Foundation"},{"name":"Japan Foundation for Pediatric Research"},{"name":"Mishima Kaiun Memorial Foundation"},{"name":"Mitsubishi Tanabe Science Foundation"},{"name":"Otsuka Science Foundation"},{"name":"Takeda Science Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathophysiology"],"abstract":"<jats:p>Autism spectrum disorder (ASD) is a neurodevelopmental disorder that includes autism, Asperger\u2019s syndrome, and pervasive developmental disorder. ASD is characterized by poor interpersonal relationships and strong attachment. The correlations between activated or inactivated gene products, which occur as a result of genetic mutations affecting neurons in ASD patients, and ASD symptoms are now of critical concern. Here, for the first time, we describe the process in which that the respective ASD-associated mutations (Arg676-to-Cys [R676C] and Ser951-to-Cys [S951C]) of semaphorin-5A (Sema5A) localize Sema5A proteins themselves around the plasma membrane in the N1E-115 cell line, a model line that can achieve neuronal morphological differentiation. The expression of each mutated construct resulted in the promotion of excessive elongation of neurite-like processes with increased differentiation protein markers; R676C was more effective than S951C. The differentiated phenotypes were very partially neutralized by an antibody, against Plexin-B3 as the specific Sema5A receptor, suggesting that the effects of Sema5A act in an autocrine manner. R676C greatly increased the activation of c-Jun N-terminal kinase (JNK), one of the signaling molecules underlying process elongation. In contrast, the blocking of JNK signaling, by a chemical JNK inhibitor or an inhibitory construct of the interaction of RhoG with Elmo1 as JNK upstream signaling molecules, recovered the excessive process elongation. These results suggest that ASD-associated mutations of Sema5A, acting through the JNK signaling cascade, lead to excessive differentiated phenotypes, and the inhibition of JNK signaling recovers them, revealing possible therapeutic targets for recovering the potential molecular and cellular phenotypes underlying certain ASD symptoms.<\/jats:p>","DOI":"10.3390\/pathophysiology30040040","type":"journal-article","created":{"date-parts":[[2023,11,27]],"date-time":"2023-11-27T03:48:17Z","timestamp":1701056897000},"page":"548-566","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["RhoG-Binding Domain of Elmo1 Ameliorates Excessive Process Elongation Induced by Autism Spectrum Disorder-Associated Sema5A"],"prefix":"10.3390","volume":"30","author":[{"given":"Miyu","family":"Okabe","sequence":"first","affiliation":[{"name":"Laboratory of Molecular Neurology, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9298-5799","authenticated-orcid":false,"given":"Yuki","family":"Miyamoto","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Neurology, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"},{"name":"Laboratory of Molecular Pharmacology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuta","family":"Ikoma","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Neurology, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikito","family":"Takahashi","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Neurology, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Remina","family":"Shirai","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Neurology, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4178-1611","authenticated-orcid":false,"given":"Mutsuko","family":"Kukimoto-Niino","sequence":"additional","affiliation":[{"name":"Laboratory for Protein Functional and Structural Biology, Center for Biosystems Dynamics Research, RIKEN, Yokohama 230-0045, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikako","family":"Shirouzu","sequence":"additional","affiliation":[{"name":"Laboratory for Protein Functional and Structural Biology, Center for Biosystems Dynamics Research, RIKEN, Yokohama 230-0045, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3618-998X","authenticated-orcid":false,"given":"Junji","family":"Yamauchi","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Neurology, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"},{"name":"Laboratory of Molecular Pharmacology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan"},{"name":"Diabetic Neuropathy Project, Tokyo Metropolitan Institute of Medical Science,Tokyo 156-8506, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,11,27]]},"reference":[{"key":"ref_1","first-page":"855","article-title":"Psychopathology, families, and culture: Autism. Child Adolesc","volume":"19","author":"Bernier","year":"2010","journal-title":"Psychiatr. Clin. N. Am."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"791","DOI":"10.3389\/fnhum.2014.00791","article-title":"Affective and cognitive empathy in adolescents with autism spectrum disorder","volume":"8","author":"Mazza","year":"2014","journal-title":"Front. Hum. Neurosci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.copsyc.2018.07.004","article-title":"The state of research on the genetics of autism spectrum disorder: Methodological, clinical and conceptual progress","volume":"27","author":"Arnett","year":"2019","journal-title":"Curr. Opin. Psychol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1007\/s40653-019-00253-5","article-title":"Trauma and autism spectrum disorder: Review, proposed treatment adaptations and future directions","volume":"12","author":"Peterson","year":"2019","journal-title":"J. Child. Adolesc. Trauma"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1186\/s13229-017-0121-4","article-title":"Environmental risk factors for autism: An evidence-based review of systematic reviews and meta-analyses","volume":"8","author":"Modabbernia","year":"2017","journal-title":"Mol. Autism."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1007\/s00401-017-1736-4","article-title":"Autism spectrum disorder: Neuropathology and animal models","volume":"134","author":"Varghese","year":"2017","journal-title":"Acta Neuropathol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Genovese, A., and Butler, M.G. (2020). Clinical assessment, genetics, and treatment approaches in autism spectrum disorder (ASD). Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21134726"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Wang, L., Wang, B., Wu, C., Wang, J., and Sun, M. (2023). Autism spectrum disorder: Neurodevelopmental risk factors, biological mechanism, and precision therapy. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24031819"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1146\/annurev.ne.17.030194.001411","article-title":"Neuronal polarity","volume":"17","author":"Craig","year":"1994","journal-title":"Annu. Rev. Neurosci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1038\/nrn918","article-title":"Breaking the neuronal sphere: Regulation of the actin cytoskeleton in neuritogenesis","volume":"3","author":"Dotti","year":"2002","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1038\/nrn2056","article-title":"Neuronal polarity: From extracellular signals to intracellular mechanisms","volume":"8","author":"Arimura","year":"2007","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1038\/nrn3379","article-title":"Neurotrophin regulation of neural circuit development and function","volume":"14","author":"Park","year":"2013","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1073\/pnas.65.4.905","article-title":"Surface movements during the growth of single explanted neurons","volume":"65","author":"Bray","year":"1970","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"203","DOI":"10.3389\/fnins.2020.00203","article-title":"Glial cell-axonal growth cone interactions in neurodevelopment and regeneration","volume":"14","author":"Rigby","year":"2020","journal-title":"Front. Neurosci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1038\/nrd4337","article-title":"Semaphorins and plexins as therapeutic targets","volume":"13","author":"Worzfeld","year":"2014","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1016\/j.tins.2008.11.006","article-title":"Axon guidance and synaptic maintenance: Preclinical markers for neurodegenerative disease and therapeutics","volume":"32","author":"Lin","year":"2009","journal-title":"Trends. Neurosci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1016\/j.conb.2020.12.014","article-title":"Semaphorins and Plexins in central nervous system patterning: The key to it all?","volume":"66","author":"Limoni","year":"2021","journal-title":"Curr. Opin. Neurobiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/bs.ctdb.2020.11.008","article-title":"New insights into the molecular mechanisms of axon guidance receptor regulation and signaling","volume":"142","author":"Zang","year":"2021","journal-title":"Curr. Top. Dev. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1016\/j.neuron.2011.06.009","article-title":"Class 5 transmembrane semaphorins control selective mammalian retinal lamination and function","volume":"71","author":"Matsuoka","year":"2011","journal-title":"Neuron"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4989","DOI":"10.1523\/JNEUROSCI.4390-03.2004","article-title":"An oligodendrocyte lineage-specific semaphorin, Sema5A, inhibits axon growth by retinal ganglion cells","volume":"24","author":"Goldberg","year":"2004","journal-title":"J. Neurosci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1159\/000096040","article-title":"Constitutional downregulation of SEMA5A expression in autism","volume":"54","author":"Melin","year":"2006","journal-title":"Neuropsychobiology"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1002\/ajmg.a.37445","article-title":"Multigenerational autosomal dominant inheritance of 5p chromosomal deletions","volume":"170","author":"Zhang","year":"2016","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1007\/s00795-017-0161-z","article-title":"Autism spectrum disorder-associated genes and the development of dentate granule cells","volume":"50","author":"Ito","year":"2017","journal-title":"Med. Mol. Morphol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"838","DOI":"10.1038\/ejhg.2015.211","article-title":"A de novo microdeletion of SEMA5A in a boy with autism spectrum disorder and intellectual disability","volume":"24","author":"Gueneau","year":"2016","journal-title":"Eur. J. Hum Genet."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2000","DOI":"10.1212\/WNL.0b013e3181e396a8","article-title":"Familial cortical myoclonic tremor with epilepsy: The third locus (FCMTE3) maps to 5p","volume":"74","author":"Depienne","year":"2010","journal-title":"Neurology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"605","DOI":"10.3389\/fgene.2019.00605","article-title":"De novo germline mutations in SEMA5A associated with infantile spasms","volume":"10","author":"Wang","year":"2019","journal-title":"Front. Genet."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1016\/j.bbr.2011.07.008","article-title":"Are Sema5a mutant mice a good model of autism? A behavioral analysis of sensory systems, emotionality and cognition","volume":"225","author":"Gunn","year":"2011","journal-title":"Behav. Brain Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e04390","DOI":"10.7554\/eLife.04390","article-title":"Semaphorin 5A inhibits synaptogenesis in early postnatal- and adult-born hippocampal dentate granule cells","volume":"3","author":"Duan","year":"2014","journal-title":"Elife"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1625","DOI":"10.1083\/jcb.141.7.1625","article-title":"Molecular dissection of the Rho-associated protein kinase (p160ROCK)-regulated neurite remodeling in neuroblastoma N1E-115 cells","volume":"141","author":"Hirose","year":"1998","journal-title":"J. Cell Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"791","DOI":"10.1016\/j.yexcr.2006.11.017","article-title":"Dock6, a Dock-C subfamily guanine nucleotide exchanger, has the dual specificity for Rac1 and Cdc42 and regulates neurite outgrowth","volume":"313","author":"Miyamoto","year":"2007","journal-title":"Exp. Cell Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/S0968-0004(00)88978-1","article-title":"Parallel signal processing among mammalian MAPKs","volume":"20","author":"Cano","year":"1995","journal-title":"Trends. Biochem. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1016\/j.bbadis.2009.12.009","article-title":"Pathological roles of MAPK signaling pathways in human diseases","volume":"1802","author":"Kim","year":"2010","journal-title":"Biochim. Biophys. Acta."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1016\/S0076-6879(06)06028-9","article-title":"Dock180-ELMO cooperation in Rac activation","volume":"406","author":"Lu","year":"2006","journal-title":"Methods Enzymol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Tam, C., Kukimoto-Niino, M., Miyata-Yabuki, Y., Tsuda, K., Mishima-Tsumagari, C., Ihara, K., Inoue, M., Yonemochi, M., Hanada, K., and Matsumoto, T. (2023). Targeting Ras-binding domain of ELMO1 by computational nanobody design. Commun. Biol., 6.","DOI":"10.1038\/s42003-023-04657-w"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"710","DOI":"10.1038\/sj.embor.7400189","article-title":"Plexin-B3 is a functional receptor for semaphorin 5A","volume":"5","author":"Artigiani","year":"2004","journal-title":"EMBO Rep."},{"key":"ref_36","first-page":"485","article-title":"Semaphorin 5A mediated cellular navigation: Connecting nervous system and cancer","volume":"1846","author":"Purohit","year":"2014","journal-title":"Biochim. Biophys. Acta."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3859","DOI":"10.1002\/ajmg.a.62440","article-title":"PLXNA2 as a candidate gene in patients with intellectual disability","volume":"185","author":"Altuame","year":"2021","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3274467","DOI":"10.1155\/2023\/3274467","article-title":"SEMA5A-PLXNB3 axis promotes PDAC liver metastasis outgrowth through enhancing the Warburg effect","volume":"2023","author":"Wang","year":"2023","journal-title":"J. Immunol. Res."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Su, S.C., Hsin, C.H., Lu, Y.T., Chuang, C.Y., Ho, Y.T., Yeh, F.L., Yang, S.F., and Lin, C.W. (2023). EF-24, a curcumin analog, inhibits cancer cell invasion in human nasopharyngeal carcinoma through transcriptional suppression of matrix metalloproteinase-9 gene expression. Cancers, 15.","DOI":"10.3390\/cancers15051552"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"876","DOI":"10.4306\/pi.2017.14.6.876","article-title":"Polymorphism in the promoter region of SEMA5A is associated with sociality traits in Korean subjects with autism apectrum disorders","volume":"14","author":"Kim","year":"2017","journal-title":"Psychiatry Investig."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"278","DOI":"10.1186\/s13046-018-0933-x","article-title":"Semaphorin 5A drives melanoma progression: Role of Bcl-2, miR-204, and c-Myb","volume":"37","author":"Valentini","year":"2018","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1088","DOI":"10.1002\/aur.2502","article-title":"PLXNA2 and LRRC40 as candidate genes in autism spectrum disorder","volume":"14","author":"Pijuan","year":"2021","journal-title":"Autism Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1016\/j.jgg.2021.03.002","article-title":"Targeted sequencing and integrative analysis of 3195 Chinese patients with neurodevelopmental disorders prioritized 26 novel candidate genes","volume":"48","author":"Wang","year":"2021","journal-title":"J. Genet. Genomics"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.pediatrneurol.2021.10.008","article-title":"Semaphorin-Plexin signaling: From axonal guidance to a new X-linked intellectual disability syndrome","volume":"126","author":"Steele","year":"2022","journal-title":"Pediatr. Neurol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Hartwig, C., Veske, A., Krejcova, S., Rosenberger, G., and Finckh, U. (2005). Plexin B3 promotes neurite outgrowth, interacts homophilically, and interacts with Rin. BMC Neurosci., 6.","DOI":"10.1186\/1471-2202-6-53"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"32436","DOI":"10.1074\/jbc.M110.120451","article-title":"Semaphorin 5A and plexin-B3 inhibit human glioma cell motility through RhoGDIa-mediated inactivation of Rac1 GTPase","volume":"285","author":"Li","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1038\/onc.2011.256","article-title":"Semaphorin 5A and plexin-B3 regulate human glioma cell motility and morphology through Rac1 and the actin cytoskeleton","volume":"31","author":"Li","year":"2012","journal-title":"Oncogene"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"5931","DOI":"10.18632\/oncotarget.23644","article-title":"Pathological and functional significance of Semaphorin-5A in pancreatic cancer progression and metastasis","volume":"9","author":"Saxena","year":"2017","journal-title":"Oncotarget"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1186\/s12935-018-0584-1","article-title":"The association of semaphorin 5A with lymph node metastasis and adverse prognosis in cervical cancer","volume":"18","author":"Xiao","year":"2018","journal-title":"Cancer Cell Int."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"112164","DOI":"10.1016\/j.celrep.2023.112164","article-title":"Plexin-B3 expression stimulates MET signaling, breast cancer stem cell specification, and lung metastasis","volume":"42","author":"Zuo","year":"2023","journal-title":"Cell Rep."}],"container-title":["Pathophysiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1873-149X\/30\/4\/40\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:30:55Z","timestamp":1760131855000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1873-149X\/30\/4\/40"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,27]]},"references-count":50,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2023,12]]}},"alternative-id":["pathophysiology30040040"],"URL":"https:\/\/doi.org\/10.3390\/pathophysiology30040040","relation":{},"ISSN":["1873-149X"],"issn-type":[{"value":"1873-149X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,11,27]]}}}