{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T20:40:08Z","timestamp":1790887208813,"version":"4.1.0"},"reference-count":50,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,10,11]],"date-time":"2021-10-11T00:00:00Z","timestamp":1633910400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["EY024932"],"award-info":[{"award-number":["EY024932"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["EY023295"],"award-info":[{"award-number":["EY023295"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["EY028106"],"award-info":[{"award-number":["EY028106"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["EY031063"],"award-info":[{"award-number":["EY031063"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Neurosci."],"abstract":"<jats:p>Optic neuritis, inflammation, and demyelination of the optic nerve (ON), is one of the most common clinical manifestations of multiple sclerosis; affected patients suffer persistent visual symptoms due to ON degeneration and secondary retinal ganglion cell (RGC) death. The mouse experimental autoimmune encephalomyelitis (EAE) model replicates optic neuritis and significant RGC soma and axon loss. Nicotinamide mononucleotide adenylyltransferases (NMNATs) are NAD<jats:sup>+<\/jats:sup>-synthetic enzymes that have been shown to be essential for axon integrity, activation of which significantly delays axonal Wallerian degeneration. NMNAT2, which is enriched in axons, has been proposed as a promising therapeutic target for axon injury-induced neurodegeneration. We therefore investigated whether activation of NMNAT2 can be used as a gene therapy strategy for neuroprotection in EAE\/optic neuritis. To avoid the confounding effects in inflammatory cells, which play important roles in EAE initiation and progression, we used an RGC-specific promoter to drive the expression of the long half-life NMNAT2 mutant in mouse RGCs <jats:italic>in vivo<\/jats:italic>. However, optical coherence tomography <jats:italic>in vivo<\/jats:italic> retina imaging did not reveal significant protection of the ganglion cell complex, and visual function assays, pattern electroretinography, and optokinetic response also showed no improvement in mice with NMNAT2 overexpression. Postmortem histological analysis of retina wholemounts and semithin sections of ON confirmed the <jats:italic>in vivo<\/jats:italic> results: NMNAT2 activation in RGCs does not provide significant neuroprotection of RGCs in EAE\/optic neuritis. Our studies suggest that a different degenerative mechanism than Wallerian degeneration is involved in autoimmune inflammatory axonopathy and that NMNAT2 may not be a major contributor to this mechanism.<\/jats:p>","DOI":"10.3389\/fncel.2021.754651","type":"journal-article","created":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T16:00:52Z","timestamp":1634140852000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["Neuronal NMNAT2 Overexpression Does Not Achieve Significant Neuroprotection in Experimental Autoimmune Encephalomyelitis\/Optic Neuritis"],"prefix":"10.3389","volume":"15","author":[{"given":"Pingting","family":"Liu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haoliang","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fang","family":"Fang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liang","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liang","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xue","family":"Feng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roopa","family":"Dalal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Hu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,10,11]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"199","DOI":"10.1097\/WCO.0000000000000327","article-title":"Optic neuritis as a phase 2 paradigm for neuroprotection therapies of multiple sclerosis: update on current trials and perspectives.","volume":"29","author":"Aktas","year":"2016","journal-title":"Curr. Opin. Neurol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"e1002472","DOI":"10.1371\/journal.pbio.1002472","article-title":"NMNAT2:HSP90 complex mediates proteostasis in proteinopathies.","volume":"14","author":"Ali","year":"2016","journal-title":"PLoS Biol."},{"key":"B3","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1093\/brain\/awu335","article-title":"Vision and vision-related outcome measures in multiple sclerosis.","author":"Balcer","year":"2015","journal-title":"Brain"},{"key":"B4","doi-asserted-by":"publisher","first-page":"36334","DOI":"10.1074\/jbc.M508660200","article-title":"Subcellular compartmentation and differential catalytic properties of the three human nicotinamide mononucleotide adenylyltransferase isoforms.","volume":"280","author":"Berger","year":"2005","journal-title":"J. Biol. Chem."},{"key":"B5","doi-asserted-by":"publisher","first-page":"264","DOI":"10.1523\/JNEUROSCI.4178-07.2008","article-title":"A transcriptome database for astrocytes, neurons, and oligodendrocytes: a new resource for understanding brain development and function.","volume":"28","author":"Cahoy","year":"2008","journal-title":"J. Neurosci."},{"key":"B6","doi-asserted-by":"publisher","first-page":"1695","DOI":"10.2353\/ajpath.2007.060677","article-title":"Elevated neuronal expression of CD200 protects Wlds mice from inflammation-mediated neurodegeneration.","volume":"170","author":"Chitnis","year":"2007","journal-title":"Am. J. Pathol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"2469","DOI":"10.1167\/iovs.14-13943","article-title":"Robust mouse pattern electroretinograms derived simultaneously from each eye using a common snout electrode.","volume":"55","author":"Chou","year":"2014","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"B8","doi-asserted-by":"publisher","first-page":"183","DOI":"10.1038\/s41583-020-0269-3","article-title":"Programmed axon degeneration: from mouse to mechanism to medicine.","volume":"21","author":"Coleman","year":"2020","journal-title":"Nat. Rev. Neurosci."},{"key":"B9","doi-asserted-by":"publisher","first-page":"731","DOI":"10.1038\/cdd.2014.164","article-title":"A rise in NAD precursor nicotinamide mononucleotide (NMN) after injury promotes axon degeneration.","volume":"22","author":"Di Stefano","year":"2015","journal-title":"Cell Death Differ."},{"key":"B10","doi-asserted-by":"publisher","first-page":"677","DOI":"10.1017\/S0952523805225166","article-title":"Independent visual threshold measurements in the two eyes of freely moving rats and mice using a virtual-reality optokinetic system.","volume":"22","author":"Douglas","year":"2005","journal-title":"Vis. Neurosci."},{"key":"B11","doi-asserted-by":"publisher","first-page":"976","DOI":"10.1111\/j.1750-3639.2010.00401.x","article-title":"Wallerian degeneration: a major component of early axonal pathology in multiple sclerosis.","volume":"20","author":"Dziedzic","year":"2010","journal-title":"Brain Pathol."},{"key":"B12","doi-asserted-by":"publisher","first-page":"9052","DOI":"10.1038\/s41598-021-88690-x","article-title":"Chronic mild and acute severe glaucomatous neurodegeneration derived from silicone oil-induced ocular hypertension.","volume":"11","author":"Fang","year":"2021","journal-title":"Sci. Rep."},{"key":"B13","doi-asserted-by":"publisher","first-page":"3319","DOI":"10.1002\/jnr.22498","article-title":"Overexpression of Wld(S) or Nmnat2 in mauthner cells by single-cell electroporation delays axon degeneration in live zebrafish.","volume":"88","author":"Feng","year":"2010","journal-title":"J. Neurosci. Res."},{"key":"B14","doi-asserted-by":"publisher","first-page":"84","DOI":"10.3389\/fneur.2012.00084","article-title":"Resveratrol neuroprotection in a chronic mouse model of multiple sclerosis.","volume":"3","author":"Fonseca-Kelly","year":"2012","journal-title":"Front. Neurol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"224","DOI":"10.1016\/j.conb.2014.05.001","article-title":"Signaling mechanisms regulating Wallerian degeneration.","volume":"27","author":"Freeman","year":"2014","journal-title":"Curr. Opin. Neurobiol."},{"key":"B16","doi-asserted-by":"publisher","first-page":"13410","DOI":"10.1523\/JNEUROSCI.1534-13.2013","article-title":"Rescue of peripheral and CNS axon defects in mice lacking NMNAT2.","volume":"33","author":"Gilley","year":"2013","journal-title":"J. Neurosci."},{"key":"B17","doi-asserted-by":"publisher","first-page":"e1000300","DOI":"10.1371\/journal.pbio.1000300","article-title":"Endogenous Nmnat2 is an essential survival factor for maintenance of healthy axons.","volume":"8","author":"Gilley","year":"2010","journal-title":"PLoS Biol."},{"key":"B18","doi-asserted-by":"publisher","first-page":"448","DOI":"10.1093\/hmg\/ddy356","article-title":"Low levels of NMNAT2 compromise axon development and survival.","volume":"28","author":"Gilley","year":"2019","journal-title":"Hum. Mol. Genet."},{"key":"B19","doi-asserted-by":"publisher","first-page":"2696","DOI":"10.1167\/iovs.10-6015","article-title":"Spermidine alleviates severity of murine experimental autoimmune encephalomyelitis.","volume":"52","author":"Guo","year":"2011","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"B20","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1016\/j.jneumeth.2017.04.003","article-title":"Consistent induction of chronic experimental autoimmune encephalomyelitis in C57BL\/6 mice for the longitudinal study of pathology and repair.","volume":"284","author":"Hasselmann","year":"2017","journal-title":"J. Neurosci. Methods"},{"key":"B21","doi-asserted-by":"publisher","first-page":"e2936","DOI":"10.1038\/cddis.2017.329","article-title":"Neuroprotection by eIF2alpha-CHOP inhibition and XBP-1 activation in EAE\/optic neuritiss.","volume":"8","author":"Huang","year":"2017","journal-title":"Cell Death Dis."},{"key":"B22","doi-asserted-by":"publisher","first-page":"9794","DOI":"10.1523\/JNEUROSCI.2116-06.2006","article-title":"Protecting axonal degeneration by increasing nicotinamide adenine dinucleotide levels in experimental autoimmune encephalomyelitis models.","volume":"26","author":"Kaneko","year":"2006","journal-title":"J. Neurosci."},{"key":"B23","doi-asserted-by":"publisher","first-page":"107710","DOI":"10.1016\/j.exer.2019.107710","article-title":"Efficient determination of axon number in the optic nerve: a stereological approach.","volume":"186","author":"Koschade","year":"2019","journal-title":"Exp. Eye Res."},{"key":"B24","doi-asserted-by":"publisher","first-page":"109","DOI":"10.3389\/fncel.2020.00109","article-title":"Longitudinal morphological and functional assessment of RGC neurodegeneration after optic nerve crush in mouse.","volume":"14","author":"Li","year":"2020","journal-title":"Front. Cell. Neurosci."},{"key":"B25","doi-asserted-by":"publisher","first-page":"251","DOI":"10.1093\/hmg\/ddr492","article-title":"CREB-activity and nmnat2 transcription are down-regulated prior to neurodegeneration, while NMNAT2 over-expression is neuroprotective, in a mouse model of human tauopathy.","volume":"21","author":"Ljungberg","year":"2012","journal-title":"Hum. Mol. Genet."},{"key":"B26","doi-asserted-by":"publisher","first-page":"1199","DOI":"10.1038\/nn770","article-title":"Wallerian degeneration of injured axons and synapses is delayed by a Ube4b\/Nmnat chimeric gene.","volume":"4","author":"Mack","year":"2001","journal-title":"Nat. Neurosci."},{"key":"B27","doi-asserted-by":"publisher","first-page":"40387","DOI":"10.1074\/jbc.M110.178913","article-title":"Expression, localization, and biochemical characterization of nicotinamide mononucleotide adenylyltransferase 2.","volume":"285","author":"Mayer","year":"2010","journal-title":"J. Biol. Chem."},{"key":"B28","doi-asserted-by":"crossref","first-page":"6214","DOI":"10.1523\/JNEUROSCI.21-16-06214.2001","article-title":"Acute neuronal apoptosis in a rat model of multiple sclerosis.","volume":"21","author":"Meyer","year":"2001","journal-title":"J. Neurosci."},{"key":"B29","doi-asserted-by":"publisher","first-page":"e14908","DOI":"10.7554\/eLife.14908","article-title":"mTORC1 is necessary but mTORC2 and GSK3beta are inhibitory for AKT3-induced axon regeneration in the central nervous system.","volume":"5","author":"Miao","year":"2016","journal-title":"Elife"},{"key":"B30","doi-asserted-by":"publisher","first-page":"133","DOI":"10.4161\/bioa.27049","article-title":"Axonal trafficking of NMNAT2 and its roles in axon growth and survival in vivo.","volume":"3","author":"Milde","year":"","journal-title":"Bioarchitecture"},{"key":"B31","doi-asserted-by":"publisher","first-page":"e1001539","DOI":"10.1371\/journal.pbio.1001539","article-title":"Subcellular localization determines the stability and axon protective capacity of axon survival factor Nmnat2.","volume":"11","author":"Milde","year":"","journal-title":"PLoS Biol."},{"key":"B32","doi-asserted-by":"publisher","first-page":"164","DOI":"10.1016\/j.exer.2015.05.008","article-title":"Electrophysiological assessment of retinal ganglion cell function.","volume":"141","author":"Porciatti","year":"2015","journal-title":"Exp. Eye Res."},{"key":"B33","doi-asserted-by":"publisher","first-page":"4611","DOI":"10.1167\/iovs.04-0541","article-title":"Rapid quantification of adult and developing mouse spatial vision using a virtual optomotor system.","volume":"45","author":"Prusky","year":"2004","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"B34","doi-asserted-by":"publisher","first-page":"50","DOI":"10.3389\/fneur.2011.00050","article-title":"Optic neuritis and retinal ganglion cell loss in a chronic murine model of multiple sclerosis.","volume":"2","author":"Quinn","year":"2011","journal-title":"Front. Neurol."},{"key":"B35","doi-asserted-by":"publisher","first-page":"4060","DOI":"10.1167\/iovs.04-0554","article-title":"Myelin\/oligodendrocyte glycoprotein-specific T-cells induce severe optic neuritis in the C57BL\/6 mouse.","volume":"45","author":"Shao","year":"2004","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"B36","doi-asserted-by":"publisher","first-page":"526","DOI":"10.1177\/1352458506070629","article-title":"Retinal ganglion cell loss induced by acute optic neuritis in a relapsing model of multiple sclerosis.","volume":"12","author":"Shindler","year":"2006","journal-title":"Mult. Scler."},{"key":"B37","doi-asserted-by":"publisher","first-page":"208","DOI":"10.1016\/j.exer.2008.05.017","article-title":"Inflammatory demyelination induces axonal injury and retinal ganglion cell apoptosis in experimental optic neuritis.","volume":"87","author":"Shindler","year":"2008","journal-title":"Exp. Eye Res."},{"key":"B38","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1186\/s12974-017-0831-8","article-title":"Relationship of acute axonal damage, Wallerian degeneration, and clinical disability in multiple sclerosis.","volume":"14","author":"Singh","year":"2017","journal-title":"J. Neuroinflammation"},{"key":"B39","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1002\/ana.20913","article-title":"How to successfully apply animal studies in experimental allergic encephalomyelitis to research on multiple sclerosis.","volume":"60","author":"Steinman","year":"2006","journal-title":"Ann. Neurol."},{"key":"B40","doi-asserted-by":"publisher","first-page":"749","DOI":"10.1002\/ana.22005","article-title":"Longitudinal study of vision and retinal nerve fiber layer thickness in multiple sclerosis.","volume":"67","author":"Talman","year":"2010","journal-title":"Ann. Neurol."},{"key":"B41","doi-asserted-by":"publisher","first-page":"83","DOI":"10.1016\/S1474-4422(13)70259-X","article-title":"Optic neuritis.","volume":"13","author":"Toosy","year":"2014","journal-title":"Lancet Neurol."},{"key":"B42","doi-asserted-by":"publisher","first-page":"10093","DOI":"10.1073\/pnas.1508337112","article-title":"Local axonal protection by WldS as revealed by conditional regulation of protein stability.","volume":"112","author":"Wang","year":"2015","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B43","doi-asserted-by":"publisher","first-page":"349","DOI":"10.1083\/jcb.200504028","article-title":"A local mechanism mediates NAD-dependent protection of axon degeneration.","volume":"170","author":"Wang","year":"2005","journal-title":"J. Cell Biol."},{"key":"B44","doi-asserted-by":"publisher","first-page":"3896","DOI":"10.1523\/JNEUROSCI.0102-20.2020","article-title":"Mouse gamma-synuclein promoter-mediated gene expression and editing in mammalian retinal ganglion cells.","volume":"40","author":"Wang","year":"2020","journal-title":"J. Neurosci."},{"key":"B45","doi-asserted-by":"publisher","first-page":"756","DOI":"10.1126\/science.aal0092","article-title":"Vitamin B3 modulates mitochondrial vulnerability and prevents glaucoma in aged mice.","volume":"355","author":"Williams","year":"2017","journal-title":"Science"},{"key":"B46","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.brainresbull.2020.11.019","article-title":"Upregulated Nmnat2 causes neuronal death and increases seizure susceptibility in temporal lobe epilepsy.","volume":"167","author":"Wu","year":"2021","journal-title":"Brain Res. Bull."},{"key":"B47","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1016\/j.neuint.2009.09.007","article-title":"Nmnat2 delays axon degeneration in superior cervical ganglia dependent on its NAD synthesis activity.","volume":"56","author":"Yan","year":"2010","journal-title":"Neurochem. Int."},{"key":"B48","doi-asserted-by":"publisher","first-page":"5891","DOI":"10.1523\/JNEUROSCI.3709-15.2016","article-title":"Rescue of glaucomatous neurodegeneration by differentially modulating neuronal endoplasmic reticulum stress molecules.","volume":"36","author":"Yang","year":"2016","journal-title":"J. Neurosci."},{"key":"B49","doi-asserted-by":"publisher","first-page":"5416","DOI":"10.1038\/ncomms6416","article-title":"The mTORC1 effectors S6K1 and 4E-BP play different roles in CNS axon regeneration.","volume":"5","author":"Yang","year":"2014","journal-title":"Nat. Commun."},{"key":"B50","doi-asserted-by":"publisher","first-page":"e45881","DOI":"10.7554\/eLife.45881","article-title":"Silicone oil-induced ocular hypertension and glaucomatous neurodegeneration in mouse.","volume":"8","author":"Zhang","year":"2019","journal-title":"Elife"}],"container-title":["Frontiers in Cellular Neuroscience"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2021.754651\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T16:00:57Z","timestamp":1634140857000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2021.754651\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,11]]},"references-count":50,"alternative-id":["10.3389\/fncel.2021.754651"],"URL":"https:\/\/doi.org\/10.3389\/fncel.2021.754651","relation":{},"ISSN":["1662-5102"],"issn-type":[{"value":"1662-5102","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,11]]},"article-number":"754651"}}