{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,18]],"date-time":"2026-06-18T19:37:28Z","timestamp":1781811448233,"version":"3.54.5"},"reference-count":32,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2023,9,20]],"date-time":"2023-09-20T00:00:00Z","timestamp":1695168000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NEI","award":["R01EY019101"],"award-info":[{"award-number":["R01EY019101"]}]},{"name":"NEI","award":["P-30 EY012576"],"award-info":[{"award-number":["P-30 EY012576"]}]},{"name":"NEI","award":["FA9550-22-1-0149"],"award-info":[{"award-number":["FA9550-22-1-0149"]}]},{"name":"NEI","award":["FA9550-16-1-0052"],"award-info":[{"award-number":["FA9550-16-1-0052"]}]},{"name":"NEI","award":["D20AC00003"],"award-info":[{"award-number":["D20AC00003"]}]},{"name":"Burns Family Audacious seed grant, Core Grant","award":["R01EY019101"],"award-info":[{"award-number":["R01EY019101"]}]},{"name":"Burns Family Audacious seed grant, Core Grant","award":["P-30 EY012576"],"award-info":[{"award-number":["P-30 EY012576"]}]},{"name":"Burns Family Audacious seed grant, Core Grant","award":["FA9550-22-1-0149"],"award-info":[{"award-number":["FA9550-22-1-0149"]}]},{"name":"Burns Family Audacious seed grant, Core Grant","award":["FA9550-16-1-0052"],"award-info":[{"award-number":["FA9550-16-1-0052"]}]},{"name":"Burns Family Audacious seed grant, Core Grant","award":["D20AC00003"],"award-info":[{"award-number":["D20AC00003"]}]},{"name":"AFOSR DURIP","award":["R01EY019101"],"award-info":[{"award-number":["R01EY019101"]}]},{"name":"AFOSR DURIP","award":["P-30 EY012576"],"award-info":[{"award-number":["P-30 EY012576"]}]},{"name":"AFOSR DURIP","award":["FA9550-22-1-0149"],"award-info":[{"award-number":["FA9550-22-1-0149"]}]},{"name":"AFOSR DURIP","award":["FA9550-16-1-0052"],"award-info":[{"award-number":["FA9550-16-1-0052"]}]},{"name":"AFOSR DURIP","award":["D20AC00003"],"award-info":[{"award-number":["D20AC00003"]}]},{"name":"AFOSR MURI","award":["R01EY019101"],"award-info":[{"award-number":["R01EY019101"]}]},{"name":"AFOSR MURI","award":["P-30 EY012576"],"award-info":[{"award-number":["P-30 EY012576"]}]},{"name":"AFOSR MURI","award":["FA9550-22-1-0149"],"award-info":[{"award-number":["FA9550-22-1-0149"]}]},{"name":"AFOSR MURI","award":["FA9550-16-1-0052"],"award-info":[{"award-number":["FA9550-16-1-0052"]}]},{"name":"AFOSR MURI","award":["D20AC00003"],"award-info":[{"award-number":["D20AC00003"]}]},{"name":"Defense Advanced Research Projects Agency (DARPA)","award":["R01EY019101"],"award-info":[{"award-number":["R01EY019101"]}]},{"name":"Defense Advanced Research Projects Agency (DARPA)","award":["P-30 EY012576"],"award-info":[{"award-number":["P-30 EY012576"]}]},{"name":"Defense Advanced Research Projects Agency (DARPA)","award":["FA9550-22-1-0149"],"award-info":[{"award-number":["FA9550-22-1-0149"]}]},{"name":"Defense Advanced Research Projects Agency (DARPA)","award":["FA9550-16-1-0052"],"award-info":[{"award-number":["FA9550-16-1-0052"]}]},{"name":"Defense Advanced Research Projects Agency (DARPA)","award":["D20AC00003"],"award-info":[{"award-number":["D20AC00003"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Corneal wound healing is a complex biological process that integrates a host of different signals to coordinate cell behavior. Upon wounding, there is the generation of an endogenous wound electric field that serves as a powerful cue to guide cell migration. Concurrently, the corneal epithelium reduces sialylated glycoforms, suggesting that sialylation plays an important role during electrotaxis. Here, we show that pretreating human telomerase-immortalized corneal epithelial (hTCEpi) cells with a sialyltransferase inhibitor, P-3FAX-Neu5Ac (3F-Neu5Ac), improves electrotaxis by enhancing directionality, but not speed. This was recapitulated using Kifunensine, which inhibits cleavage of mannoses and therefore precludes sialylation on N-glycans. We also identified that 3F-Neu5Ac enhanced the responsiveness of the hTCEpi cell population to the electric field and that pretreated hTCEpi cells showed increased directionality even at low voltages. Furthermore, when we increased sialylation using N-azidoacetylmannosamine-tetraacylated (Ac4ManNAz), hTCEpi cells showed a decrease in both speed and directionality. Importantly, pretreating enucleated eyes with 3F-Neu5Ac significantly improved re-epithelialization in an ex vivo model of a corneal injury. Finally, we show that in hTCEpi cells, sialylation is increased by growth factor deprivation and reduced by PDGF-BB. Taken together, our results suggest that during corneal wound healing, reduced sialylated glycoforms enhance electrotaxis and re-epithelialization, potentially opening new avenues to promote corneal wound healing.<\/jats:p>","DOI":"10.3390\/ijms241814327","type":"journal-article","created":{"date-parts":[[2023,9,20]],"date-time":"2023-09-20T21:26:32Z","timestamp":1695245192000},"page":"14327","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Reducing Sialylation Enhances Electrotaxis of Corneal Epithelial Cells"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-9619-5791","authenticated-orcid":false,"given":"Bryan","family":"Le","sequence":"first","affiliation":[{"name":"Department of Ophthalmology, University of California, Davis, CA 95616, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kan","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, University of California, Davis, CA 95616, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chelsea","family":"Brown","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, University of California, Davis, CA 95616, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2966-1251","authenticated-orcid":false,"given":"Brian","family":"Reid","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, University of California, Davis, CA 95616, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amin","family":"Cressman","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Human Anatomy, University of California, Davis, CA 95817, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2500-3035","authenticated-orcid":false,"given":"Min","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, University of California, Davis, CA 95616, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1086-2646","authenticated-orcid":false,"given":"Fernando A.","family":"Fierro","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Human Anatomy, University of California, Davis, CA 95817, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,9,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1089\/wound.2013.0442","article-title":"The Electrical Response to Injury: Molecular Mechanisms and Wound Healing","volume":"3","author":"Reid","year":"2014","journal-title":"Adv. Wound Care"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Liang, Y., Tian, H., Liu, J., Lv, Y., Wang, Y., Zhang, J., and Huang, Y. (2020). Application of stable continuous external electric field promotes wound healing in pig wound model. Bioelectrochemistry, 135.","DOI":"10.1016\/j.bioelechem.2020.107578"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"13577","DOI":"10.1073\/pnas.202235299","article-title":"Electrical cues regulate the orientation and frequency of cell division and the rate of wound healing in vivo","volume":"99","author":"Song","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1096\/fj.04-2325com","article-title":"Wound healing in rat cornea: The role of electric currents","volume":"19","author":"Reid","year":"2005","journal-title":"FASEB J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1038\/nrm3383","article-title":"Vertebrate protein glycosylation: Diversity, synthesis and function","volume":"13","author":"Moremen","year":"2012","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3109","DOI":"10.1016\/j.cell.2021.04.023","article-title":"Small RNAs are modified with N-glycans and displayed on the surface of living cells","volume":"184","author":"Flynn","year":"2021","journal-title":"Cell"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Ying, L., Xu, J., Han, D., Zhang, Q., and Hong, Z. (2022). The Applications of Metabolic Glycoengineering. Front. Cell Dev. Biol., 10.","DOI":"10.3389\/fcell.2022.840831"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Templeton, K., Ramos, M., Rose, J., Le, B., Zhou, Q., Cressman, A., Ferreyra, S., Lebrilla, C.B., and Fierro, F.A. (2021). Mesenchymal Stromal Cells Regulate Sialylations of N-Glycans, Affecting Cell Migration and Survival. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22136868"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1189\/jlb.2VMA0915-432RR","article-title":"Distinct CCR7 glycosylation pattern shapes receptor signaling and endocytosis to modulate chemotactic responses","volume":"99","author":"Hauser","year":"2016","journal-title":"J. Leukoc. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"6199","DOI":"10.1039\/C9SC01360A","article-title":"Identification of potential sialic acid binding proteins on cell membranes by proximity chemical labeling","volume":"10","author":"Li","year":"2019","journal-title":"Chem. Sci."},{"key":"ref_11","first-page":"2277","article-title":"Alpha-2,3 sialylation differentiate the limbal and corneal epithelial cell phenotypes","volume":"36","author":"Wolosin","year":"1995","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"14015","DOI":"10.1074\/jbc.270.23.14015","article-title":"Neolactoglycosphingolipids, potential mediators of corneal epithelial cell migration","volume":"270","author":"Panjwani","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1076\/ceyr.19.3.212.5317","article-title":"Corneal wound-associated glycoconjugates analyzed by lectin histochemistry","volume":"19","author":"Sweatt","year":"1999","journal-title":"Curr. Eye Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1093\/glycob\/cwp133","article-title":"Analysis of differential expression of glycosyltransferases in healing corneas by glycogene microarrays","volume":"20","author":"Saravanan","year":"2010","journal-title":"Glycobiology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e00516-21","DOI":"10.1128\/IAI.00516-21","article-title":"Surface Glycans Regulate Salmonella Infection-Dependent Directional Switch in Macrophage Galvanotaxis Independent of NanH","volume":"90","author":"Sun","year":"2022","journal-title":"Infect. Immun."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1038\/nchembio.999","article-title":"Global metabolic inhibitors of sialyl- and fucosyltransferases remodel the glycome","volume":"8","author":"Rillahan","year":"2012","journal-title":"Nat. Chem. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Alonso-Garcia, V., Chaboya, C., Li, Q., Le, B., Congleton, T.J., Florez, J., Tran, V., Liu, G.Y., Yao, W., and Lebrilla, C.B. (2020). High Mannose N-Glycans Promote Migration of Bone-Marrow-Derived Mesenchymal Stromal Cells. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21197194"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1002\/jcp.22028","article-title":"Regulation of the membrane mucin Muc4 in corneal epithelial cells by proteosomal degradation and TGF-beta","volume":"223","author":"Lomako","year":"2010","journal-title":"J. Cell Physiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"15599","DOI":"10.1016\/S0021-9258(18)55439-9","article-title":"Kifunensine, a potent inhibitor of the glycoprotein processing mannosidase I","volume":"265","author":"Elbein","year":"1990","journal-title":"J. Biol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1382","DOI":"10.1093\/glycob\/cwp115","article-title":"Metabolic glycoengineering: Sialic acid and beyond","volume":"19","author":"Du","year":"2009","journal-title":"Glycobiology"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.exer.2015.06.010","article-title":"Galectin-3 enhances extracellular matrix associations and wound healing in monkey corneal epithelium","volume":"137","author":"Fujii","year":"2015","journal-title":"Exp. Eye Res."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Holdbrooks, A.T., Ankenbauer, K.E., Hwang, J., and Bellis, S.L. (2020). Regulation of inflammatory signaling by the ST6Gal-I sialyltransferase. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0241850"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1269","DOI":"10.1002\/cphy.c110044","article-title":"Modulation of voltage-gated ion channels by sialylation","volume":"2","author":"Ednie","year":"2012","journal-title":"Compr. Physiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"16123","DOI":"10.1074\/jbc.RA119.007903","article-title":"N-Glycosylation of the voltage-gated sodium channel beta2 subunit is required for efficient trafficking of Na(V)1.5\/beta2 to the plasma membrane","volume":"294","author":"Cortada","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1002\/cm.20227","article-title":"Electric field-induced polarization of charged cell surface proteins does not determine the direction of galvanotaxis","volume":"64","author":"Finkelstein","year":"2007","journal-title":"Cell Motil. Cytoskelet."},{"key":"ref_26","unstructured":"Ajit, V., Richard, D., Cummings, J.D., Esko, P.S., Gerald, W.H., Markus, A., Debra, M., Taroh, K., Nicolle, H., and Packer, J.H. (2022). Essentials of Glycobiology, Cold Springs Harbor Laboratory Press. [4th ed.]."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Sun, Y., Reid, B., Ferreira, F., Luxardi, G., Ma, L., Lokken, K.L., Zhu, K., Xu, G., Sun, Y., and Ryzhuk, V. (2019). Infection-generated electric field in gut epithelium drives bidirectional migration of macrophages. PLoS Biol., 17.","DOI":"10.1371\/journal.pbio.3000044"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"9131","DOI":"10.1096\/fj.201802657R","article-title":"Expression of integrins to control migration direction of electrotaxis","volume":"33","author":"Zhu","year":"2019","journal-title":"FASEB J."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Do, K.K., Wang, F., Lu, X., Liu, J.Y., Li, C., Ceresa, B.P., Zhang, L., Dean, D.C., and Liu, Y. (2023). Zeb1 facilitates corneal epithelial wound healing by maintaining corneal epithelial cell viability and mobility. Commun. Biol., 6.","DOI":"10.1038\/s42003-023-04831-0"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1016\/j.bpj.2010.03.063","article-title":"Role of altered sialylation of the I-like domain of beta1 integrin in the binding of fibronectin to beta1 integrin: Thermodynamics and conformational analyses","volume":"99","author":"Pan","year":"2010","journal-title":"Biophys. J."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.lfs.2016.02.071","article-title":"Effect of alpha 2,6 sialylation on integrin-mediated adhesion of breast cancer cells to fibronectin and collagen IV","volume":"149","author":"Yuan","year":"2016","journal-title":"Life Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1479","DOI":"10.1038\/nprot.2007.205","article-title":"Application of direct current electric fields to cells and tissues in vitro and modulation of wound electric field in vivo","volume":"2","author":"Song","year":"2007","journal-title":"Nat. Protoc."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/18\/14327\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:53:36Z","timestamp":1760129616000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/18\/14327"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,20]]},"references-count":32,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2023,9]]}},"alternative-id":["ijms241814327"],"URL":"https:\/\/doi.org\/10.3390\/ijms241814327","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,20]]}}}