{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T00:48:11Z","timestamp":1785372491162,"version":"3.55.0"},"reference-count":58,"publisher":"National Academy of Sciences","issue":"18","license":[{"start":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T00:00:00Z","timestamp":1777248000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000053","name":"HHS | NIH | National Eye Institute","doi-asserted-by":"publisher","award":["R01 EY032519-01A1"],"award-info":[{"award-number":["R01 EY032519-01A1"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Republic of Turkiye","award":["YLSY"],"award-info":[{"award-number":["YLSY"]}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2026,5,5]]},"abstract":"<jats:p>Retinal ganglion cell (RGC) axons form the optic nerve (ON). Numerous age-related ON diseases, including glaucoma, the second most common cause of worldwide blindness, result from multiple RGC stressors. Nearly all ON astrocytes in the optic nerve head (ONH): the junctional region between the ON and the retina in young-adult rodents expresses the homeodomain only (Hopx) protein. Hopx(+) ONH astrocytes are depleted during aging. ONH primary cultures which include Hopx(+) astrocytes secrete extracellular vesicles (ONH-EVs) which selectively enhance RGC survival and neurite extension in culture, while extracellular vesicles (EVs) secreted from distal ON cultures lacking Hopx(+) astrocytes do not. ONH-EVs also enhance RGC survival in vivo in a rodent model of glaucoma. Combining rat ONH single-cell (scRNA-seq) sequencing with EV proteomic analysis, we identified ONH-Hopx(+) astrocyte secreted factors. We interrogated the online Broad institute scRNA-seq database for rat RGC gene expression in control animals and following rodent ON crush, an RGC stress model, to correlate ONH-astrocyte secreted factors with RGC gene expression changes. Following stress, RGCs upregulate the complementary pathways involving Hopx(+) astrocytic-associated factors, suggesting reciprocal communication. Using a highly selective transgenic Hopx-cre ONH knockdown strategy, we demonstrate that eliminating Hopx(+) astrocytes also results in upregulation of RGC stress responses. Our results implicate age-related loss of young ONH-astrocytes as a crucial factor in the development of age-related optic nerve diseases, and discuss replacing ONH associated factors as a paradigm shift for ON disease treatment.<\/jats:p>","DOI":"10.1073\/pnas.2515277123","type":"journal-article","created":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T19:05:09Z","timestamp":1777316709000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":1,"title":["Hopx(+) optic nerve head-astrocytes counter neuronal stress and glaucoma damage"],"prefix":"10.1073","volume":"123","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7758-0136","authenticated-orcid":false,"given":"Steven L.","family":"Bernstein","sequence":"first","affiliation":[{"name":"Department of Ophthalmology and Visual Sciences, Lab of Molecular Research, University of Maryland at Baltimore School of Medicine","place":["Baltimore"]},{"name":"Department of Neurobiology and Anatomy, Lab of Molecular Research, University of Maryland at Baltimore School of Medicine","place":["Baltimore"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-5594-3323","authenticated-orcid":false,"given":"Zara","family":"Mehrabian","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology and Visual Sciences, Lab of Molecular Research, University of Maryland at Baltimore School of Medicine","place":["Baltimore"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-5850-8700","authenticated-orcid":false,"given":"Yan","family":"Guo","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology and Visual Sciences, Lab of Molecular Research, University of Maryland at Baltimore School of Medicine","place":["Baltimore"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-8523-0715","authenticated-orcid":false,"given":"Jean","family":"Jouffroy","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology and Visual Sciences, Lab of Molecular Research, University of Maryland at Baltimore School of Medicine","place":["Baltimore"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5855-0097","authenticated-orcid":false,"given":"Ben","family":"Mead","sequence":"additional","affiliation":[{"name":"Exosome and Retinal Research Group, School of Optometry and Vision Sciences, University of Wales","place":["Cardiff, United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3042-8502","authenticated-orcid":false,"given":"Stanislav","family":"Tomarev","sequence":"additional","affiliation":[{"name":"Retinal Ganglion Cell Biology Lab, National Eye Institute, National Institutes of Health","place":["Bethesda"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1825-5373","authenticated-orcid":false,"given":"Esmahan","family":"Durmaz","sequence":"additional","affiliation":[{"name":"Exosome and Retinal Research Group, School of Optometry and Vision Sciences, University of Wales","place":["Cardiff, United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8637-4465","authenticated-orcid":false,"given":"Jonathan A.","family":"Epstein","sequence":"additional","affiliation":[{"name":"Department of Medicine, Perelman School of Medicine, University of Pennsylvania","place":["Philadelphia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-6425-3544","authenticated-orcid":false,"given":"Adam","family":"Kronk","sequence":"additional","affiliation":[{"name":"Office of Research and Development, University of Maryland Ventures","place":["Baltimore"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1338-8476","authenticated-orcid":false,"given":"Seth","family":"Blackshaw","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Johns Hopkins University School of Medicine","place":["Baltimore"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7666-3842","authenticated-orcid":false,"given":"Thanh","family":"Hoang","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Michigan Neuroscience Institute, University of Michigan","place":["Ann Arbor"]},{"name":"Department of Cell and Developmental Biology, Michigan Neuroscience Institute, University of Michigan","place":["Ann Arbor"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2026,4,27]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.preteyeres.2011.11.002"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.preteyeres.2021.100953"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.exer.2017.02.006"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ophtha.2018.08.025"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1167\/iovs.13-12327"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.optom.2017.06.002"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1136\/bjo.2005.081224"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(17)31469-1"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.2174\/1874609811104020110"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00347-016-0430-6"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajo.2012.02.024"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1463-5224.2007.00504.x"},{"key":"e_1_3_4_13_2","first-page":"2197","article-title":"Localization of the neurotrophin receptor TrkB in normal and glaucomatous monkey optic nerve heads","volume":"38","author":"Pease M. E.","year":"2000","unstructured":"M. E. Pease, S. J. MacKinnon, D. J. Zack, H. A. Quigley, Localization of the neurotrophin receptor TrkB in normal and glaucomatous monkey optic nerve heads. Invest. Ophthalmol. Vis. Sci. 38, 2197\u20132197 (2000).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.exer.2009.02.015"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0079-6123(08)01136-9"},{"key":"e_1_3_4_16_2","first-page":"760","article-title":"CNTF, not other trophic factors, promotes axonal regeneration of axotomized retinal ganglion cells in adult hamsters","volume":"40","author":"Cui Q.","year":"1999","unstructured":"Q. Cui, Q. Lu, K. F. So, H. K. Yip, CNTF, not other trophic factors, promotes axonal regeneration of axotomized retinal ganglion cells in adult hamsters. Invest. Ophthalmol. Vis. Sci. 40, 760\u2013766 (1999).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.12659\/MSM.891343"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pneurobio.2018.05.004"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature23282"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1089\/neu.2008.0656"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2001858117"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.02.010"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2018.08.006"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1002\/sctm.16-0428"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.4103\/1673-5374.226392"},{"key":"e_1_3_4_26_2","unstructured":"S. Bernstein Rat optic nerve head extracellular vesicle analysis. PRIDE. https:\/\/www.ebi.ac.uk\/pride\/archive\/projects\/PXD058589. Deposited 5 December 2024."},{"key":"e_1_3_4_27_2","unstructured":"S. Bernstein . Optic nerve head-neural progenitor cells inhibit neuronal stress and glaucoma. NCBI GEO. https:\/\/www.ncbi.nlm.nih.gov\/geo\/query\/acc.cgi?acc=GSE282446. Deposited 20 November 2024."},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1213214"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.xgen.2022.100164"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2302089120"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.abb8598"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1097\/01.WCB.0000084252.20114.BE"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1002\/glia.22588"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0304-3940(03)00407-5"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1634\/stemcells.2007-0221"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.19-13-05245.1999"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.21307\/ane-2019-038"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2019.11.006"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1210\/endrev\/bnaa007"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1068428"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.3389\/fneur.2022.931523"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1038\/cddis.2017.338"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.3109\/00016489.2015.1012276"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-023-06935-3"},{"key":"e_1_3_4_45_2","first-page":"153","article-title":"The cellular basis of neurovascular metabolic coupling","volume":"102","author":"Paemeleire K.","year":"2002","unstructured":"K. Paemeleire, The cellular basis of neurovascular metabolic coupling. Acta Neurol. Belg. 102, 153\u2013157 (2002).","journal-title":"Acta Neurol. Belg."},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2306153120"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1007\/s11906-014-0512-2"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.3390\/cells14040289"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1359-6446(04)03333-1"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcpro.2023.100654"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-018-0467-6"},{"key":"e_1_3_4_52_2","doi-asserted-by":"crossref","unstructured":"Y. Guo Z. Mehrabian J. Milbrandt A. DiAntonio S. L. Bernstein Synergistic protection of Retinal Ganglion Cells (RGCs) by SARM1 inactivation with CNTF in a rodent model of nonarteritic anterior ischemic optic neuropathy. Cells 13 202 (2024).","DOI":"10.3390\/cells13030202"},{"key":"e_1_3_4_53_2","doi-asserted-by":"crossref","unstructured":"K. S. Matho Genetic dissection of glutamatergic neuron system in cerebral cortex. Nature 598 182\u2013187 (2021).","DOI":"10.1038\/s41586-021-03955-9"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.3390\/ijms232415604"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-24085-9"},{"key":"e_1_3_4_56_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.4096"},{"key":"e_1_3_4_57_2","first-page":"2033","article-title":"Establishing primary adult fibroblast cultures from rodents","volume":"44","author":"Seluanov A.","year":"2010","unstructured":"A. Seluanov, A. Vaidya, V. Gorbunova, Establishing primary adult fibroblast cultures from rodents. J. Vis. Exp. 44, 2033 (2010).","journal-title":"J. Vis. Exp."},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1167\/iovs.09-3947"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2515277123","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T19:05:18Z","timestamp":1777316718000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2515277123"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,27]]},"references-count":58,"journal-issue":{"issue":"18","published-print":{"date-parts":[[2026,5,5]]}},"alternative-id":["10.1073\/pnas.2515277123"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2515277123","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,27]]},"assertion":[{"value":"2025-06-15","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-02-22","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-04-27","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2515277123"}}