{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T05:51:19Z","timestamp":1782280279044,"version":"3.54.5"},"posted":{"date-parts":[[2025,6,11]]},"group-title":"Cell Biology","reference-count":43,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,6,11]],"date-time":"2025-06-11T00:00:00Z","timestamp":1749600000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"NIAMS\/NIH","award":["R37AR047709"],"award-info":[{"award-number":["R37AR047709"]}]},{"name":"NIAMS\/NIH","award":["RO1AR081322"],"award-info":[{"award-number":["RO1AR081322"]}]},{"name":"NIAMS\/NIH","award":["R01AR076428"],"award-info":[{"award-number":["R01AR076428"]}]},{"name":"NIAMS\/NIH","award":["P30AR079200"],"award-info":[{"award-number":["P30AR079200"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,6,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Purpose<\/jats:title>\n                  <jats:p>To investigate whether KRT6A or KRT17 label self-renewing stem cells (SCs) in adult Meibomian gland (MG) and palpebral conjunctiva (PC) homeostasis and explore the mechanisms that regulate their expression in these tissues.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    KRT6A and KRT17 expression in adult mouse MG and PC was examined by single nucleus RNA sequencing and immunofluorescence (IF). Bromodeoxyuridine pulse-chase assays were conducted to assess the turnover rate of PC progenitor cells. Lineage tracing experiments were performed using\n                    <jats:italic>\n                      Krt6a-Cre\n                      <jats:sup>ERT2<\/jats:sup>\n                    <\/jats:italic>\n                    and\n                    <jats:italic>\n                      Krt17-Cre\n                      <jats:sup>ERT2<\/jats:sup>\n                    <\/jats:italic>\n                    mice carrying\n                    <jats:italic>\n                      Rosa26R\n                      <jats:sup>nTnG<\/jats:sup>\n                    <\/jats:italic>\n                    or\n                    <jats:italic>\n                      Rosa26R\n                      <jats:sup>mTmG<\/jats:sup>\n                    <\/jats:italic>\n                    reporter. As Hedgehog (Hh) signaling, the histone deacetylase HDAC3, and the transcription factor KLF4, regulate KRT6A and KRT17 in other contexts, IF was conducted to assess the in vivo effects of overexpression of the Hh pathway activator GLI2\u0394N, and inducible epithelial deletion of\n                    <jats:italic>Hdac3<\/jats:italic>\n                    or\n                    <jats:italic>Klf4<\/jats:italic>\n                    , on KRT6A and KRT17 expression in the MG and PC.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    KRT6A and KRT17 are primarily expressed in the MG central duct and ductules. KRT6A also shows robust expression in PC. MG and PC epithelial progenitor cells turn over within 14 days. Lineage tracing indicated that\n                    <jats:italic>Krt17<\/jats:italic>\n                    labels self-renewing SCs in the MG whereas\n                    <jats:italic>Krt6a<\/jats:italic>\n                    labels SCs in the PC. GLI2\u0394N overexpression induced ectopic KRT17 expression in MG acini and PC but did not affect KRT6A expression in either MG or PC.\n                    <jats:italic>Hdac3<\/jats:italic>\n                    deficiency caused expanded expression of KRT6A and KRT17 in MG acini and ectopic KRT17 expression in PC.\n                    <jats:italic>Klf4<\/jats:italic>\n                    deletion resulted in ectopic KRT17 expression in PC but did not influence KRT6A expression in MG or PC.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Krt6a<\/jats:italic>\n                    - and\n                    <jats:italic>Krt17<\/jats:italic>\n                    -expressing cells contribute to adult PC and MG homeostasis, respectively. KRT17 expression is enhanced by GLI2\u0394N, and suppressed by HDAC3 and KLF4, whereas KRT6A expression is controlled only by HDAC3.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.06.10.658923","type":"posted-content","created":{"date-parts":[[2025,6,11]],"date-time":"2025-06-11T11:15:16Z","timestamp":1749640516000},"source":"Crossref","is-referenced-by-count":1,"title":["KRT6A and KRT17 mark distinct stem cell populations in the adult palpebral conjunctiva and Meibomian gland"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1148-4162","authenticated-orcid":false,"given":"Xuming","family":"Zhu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingang","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David M.","family":"Owens","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8792-6890","authenticated-orcid":false,"given":"Sarah E.","family":"Millar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025061306550840000_2025.06.10.658923v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.exer.2020.108115"},{"key":"2025061306550840000_2025.06.10.658923v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.exer.2022.109101"},{"key":"2025061306550840000_2025.06.10.658923v1.3","doi-asserted-by":"publisher","DOI":"10.1007\/s40123-023-00669-1"},{"key":"2025061306550840000_2025.06.10.658923v1.4","doi-asserted-by":"publisher","DOI":"10.1097\/ICO.0000000000000815"},{"key":"2025061306550840000_2025.06.10.658923v1.5","doi-asserted-by":"publisher","DOI":"10.1167\/iovs.10-6997c"},{"key":"2025061306550840000_2025.06.10.658923v1.6","unstructured":"Wolff E , Bron AJ , Tripathi RC , Tripathi BJ. Anatomy of the eye and orbit. 8th ed: Chapman & Hall Medical; 1997."},{"key":"2025061306550840000_2025.06.10.658923v1.7","doi-asserted-by":"publisher","DOI":"10.1186\/s13287-019-1165-5"},{"key":"2025061306550840000_2025.06.10.658923v1.8","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-062917-012341"},{"key":"2025061306550840000_2025.06.10.658923v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2016.07.010"},{"key":"2025061306550840000_2025.06.10.658923v1.10","unstructured":"Shumway CL , Motlagh M , Wade M. Anatomy , Head and Neck , Eye Conjunctiva . StatPearls. Treasure Island (FL)2024."},{"key":"2025061306550840000_2025.06.10.658923v1.11","doi-asserted-by":"publisher","DOI":"10.1016\/j.preteyeres.2016.04.005"},{"key":"2025061306550840000_2025.06.10.658923v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.jtos.2021.03.003"},{"key":"2025061306550840000_2025.06.10.658923v1.13","doi-asserted-by":"publisher","DOI":"10.1167\/iovs.04-0356"},{"key":"2025061306550840000_2025.06.10.658923v1.14","doi-asserted-by":"publisher","DOI":"10.1167\/iovs.14-16266"},{"key":"2025061306550840000_2025.06.10.658923v1.15","first-page":"236","article-title":"Label-retaining cells are preferentially located in fornical epithelium: implications on conjunctival epithelial homeostasis","volume":"36","year":"1995","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"2025061306550840000_2025.06.10.658923v1.16","first-page":"301","article-title":"Phorbol ester preferentially stimulates mouse fornical conjunctival and limbal epithelial cells to proliferate in vivo","volume":"39","year":"1998","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"2025061306550840000_2025.06.10.658923v1.17","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2024.108805"},{"key":"2025061306550840000_2025.06.10.658923v1.18","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1008206"},{"key":"2025061306550840000_2025.06.10.658923v1.19","doi-asserted-by":"publisher","DOI":"10.1038\/ng0795-363"},{"key":"2025061306550840000_2025.06.10.658923v1.20","doi-asserted-by":"publisher","DOI":"10.1038\/ng0395-273"},{"key":"2025061306550840000_2025.06.10.658923v1.21","doi-asserted-by":"crossref","unstructured":"Lu Z , Xie Y , Huang H , Jiang K , Zhou B , Wang F , et al. Hair follicle stem cells regulate retinoid metabolism to maintain the self-renewal niche for melanocyte stem cells. Elife, 9 (2020), pp.","DOI":"10.7554\/eLife.52712"},{"key":"2025061306550840000_2025.06.10.658923v1.22","doi-asserted-by":"publisher","DOI":"10.1046\/j.1523-1747.2000.00986.x"},{"key":"2025061306550840000_2025.06.10.658923v1.23","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2133.1998.02499.x"},{"key":"2025061306550840000_2025.06.10.658923v1.24","doi-asserted-by":"publisher","DOI":"10.1016\/j.jid.2015.12.021"},{"key":"2025061306550840000_2025.06.10.658923v1.25","doi-asserted-by":"crossref","unstructured":"Zhang X , Yin M , Zhang LJ . Keratin 6, 16 and 17-Critical Barrier Alarmin Molecules in Skin Wounds and Psoriasis. Cells, 8 (2019), pp.","DOI":"10.3390\/cells8080807"},{"key":"2025061306550840000_2025.06.10.658923v1.26","doi-asserted-by":"publisher","DOI":"10.2353\/ajpath.2008.071089"},{"key":"2025061306550840000_2025.06.10.658923v1.27","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1221804"},{"key":"2025061306550840000_2025.06.10.658923v1.28","doi-asserted-by":"publisher","DOI":"10.1101\/gad.333674.119"},{"key":"2025061306550840000_2025.06.10.658923v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.jtos.2024.04.005"},{"key":"2025061306550840000_2025.06.10.658923v1.30","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-56907-6"},{"key":"2025061306550840000_2025.06.10.658923v1.31","doi-asserted-by":"crossref","unstructured":"Zhu X , Xu M , Portal C , Lin Y , Ferdinand A , Peng T , et al. Identification of Meibomian gland stem cell populations and mechanisms of aging. bioRxiv, (2024), pp. 2024.2008.2009.607015","DOI":"10.1101\/2024.08.09.607015"},{"key":"2025061306550840000_2025.06.10.658923v1.32","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2013.04.026"},{"key":"2025061306550840000_2025.06.10.658923v1.33","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.5052-10.2011"},{"key":"2025061306550840000_2025.06.10.658923v1.34","doi-asserted-by":"publisher","DOI":"10.1046\/j.1523-1747.2000.00144.x"},{"key":"2025061306550840000_2025.06.10.658923v1.35","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.152238499"},{"key":"2025061306550840000_2025.06.10.658923v1.36","doi-asserted-by":"publisher","DOI":"10.1172\/JCI46307"},{"key":"2025061306550840000_2025.06.10.658923v1.37","doi-asserted-by":"publisher","DOI":"10.1016\/s0016-5085(03)80453-5"},{"key":"2025061306550840000_2025.06.10.658923v1.38","doi-asserted-by":"publisher","DOI":"10.3390\/cimb46080508"},{"key":"2025061306550840000_2025.06.10.658923v1.39","doi-asserted-by":"publisher","DOI":"10.1152\/ajprenal.00538.2014"},{"key":"2025061306550840000_2025.06.10.658923v1.40","doi-asserted-by":"publisher","DOI":"10.1016\/j.jid.2021.05.026"},{"key":"2025061306550840000_2025.06.10.658923v1.41","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2021.04.003"},{"key":"2025061306550840000_2025.06.10.658923v1.42","doi-asserted-by":"crossref","unstructured":"Ishii R , Yanagisawa H , Sada A . Defining compartmentalized stem cell populations with distinct cell division dynamics in the ocular surface epithelium. Development, 147 (2020), pp.","DOI":"10.1242\/dev.197590"},{"key":"2025061306550840000_2025.06.10.658923v1.43","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.00846-06"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.06.10.658923","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T05:00:42Z","timestamp":1782277242000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.06.10.658923"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,11]]},"references-count":43,"URL":"https:\/\/doi.org\/10.1101\/2025.06.10.658923","relation":{"is-preprint-of":[{"id-type":"doi","id":"10.3390\/cells14241979","asserted-by":"subject"}]},"subject":[],"published":{"date-parts":[[2025,6,11]]},"subtype":"preprint"}}