{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T17:09:24Z","timestamp":1790096964660,"version":"4.0.1"},"posted":{"date-parts":[[2026,9,18]]},"group-title":"Cell Biology","reference-count":32,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"NIH\/NIDDK","award":["R01DK136987"],"award-info":[{"award-number":["R01DK136987"]}]},{"name":"NIH\/NIDDK","award":["F31DK139760"],"award-info":[{"award-number":["F31DK139760"]}]},{"name":"NIH\/NIAID","award":["R01AI184785"],"award-info":[{"award-number":["R01AI184785"]}]},{"name":"NIH\/NCI","award":["F31CA294914"],"award-info":[{"award-number":["F31CA294914"]}]},{"name":"NIH\/NCI","award":["P30CA006927"],"award-info":[{"award-number":["P30CA006927"]}]},{"name":"NIH\/NCI","award":["U54CA221704"],"award-info":[{"award-number":["U54CA221704"]}]},{"name":"NIH\/NIGMS","award":["T32GM142606"],"award-info":[{"award-number":["T32GM142606"]}]},{"name":"NIH\/NIGMS","award":["R01GM155497"],"award-info":[{"award-number":["R01GM155497"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Mitochondria are increasingly recognized as regulators of cellular differentiation, but their role in esophageal epithelial homeostasis remains poorly understood. Here, we investigated mitochondrial remodeling during esophageal epithelial differentiation and whether mitochondrial depletion contributes to the acquisition of the differentiated phenotype. Mitochondrial abundance was assessed across normal human esophageal epithelium and in non-transformed immortalized human esophageal epithelial cells (EPC2-hTERT) using two differentiation models. Mitochondrial architecture was quantified, and the functional role of mitochondrial abundance was examined using doxycycline-inducible depletion of transcription factor A, mitochondrial (TFAM). Mitochondrial abundance progressively decreased from basal to superficial compartments of normal human esophageal epithelium and during\n                  <jats:italic>in vitro<\/jats:italic>\n                  differentiation, accompanied by fragmentation and remodeling of the mitochondrial network. TFAM depletion reduced mitochondrial abundance and increased squamous differentiation markers, indicating that mitochondrial depletion was sufficient to promote differentiation. Analysis of candidate mitochondrial clearance pathways identified Bcl2-interacting protein 3-like (\n                  <jats:italic>BNIP3L<\/jats:italic>\n                  )\/NIX (NIP3-like protein X) as preferentially associated with differentiated epithelial cells and induced during differentiation. NIX was associated with mitochondria and increased as mitochondrial abundance declined. NIX depletion prevented differentiation-associated mitochondrial depletion and attenuated differentiation marker induction. In human esophageal epithelium, NIX expression increased across the basal-to-suprabasal compartment before declining superficially.\n                  <jats:italic>BNIP3L<\/jats:italic>\n                  expression was reduced in active eosinophilic esophagitis (EoE) and increased following corticosteroid-associated remission. Interleukin-13 similarly reduced\n                  <jats:italic>BNIP3L<\/jats:italic>\n                  expression and suppressed epithelial differentiation in EPC2-hTERT cells. These findings identify NIX-associated mitochondrial remodeling as an important component of esophageal epithelial differentiation and suggest that disruption of mitochondrial quality control may contribute to impaired epithelial differentiation in EoE.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>New and Noteworthy<\/jats:title>\n                  <jats:p>Although impaired esophageal epithelial differentiation is associated with various esophageal diseases, mechanisms regulating differentiation under homeostasis remain incompletely understood. We show that mitochondria are progressively cleared during esophageal epithelial differentiation and that mitochondrial depletion is sufficient to promote differentiation. BNIP3L\/NIX regulates mitochondrial clearance during this process. Moreover, BNIP3L expression is reduced in active eosinophilic esophagitis (EoE) and restored with remission, linking disrupted mitochondrial quality control to epithelial dysfunction in EoE.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.09.17.752311","type":"posted-content","created":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T14:00:19Z","timestamp":1789740019000},"source":"Crossref","is-referenced-by-count":0,"title":["NIX-associated mitochondrial remodeling contributes to esophageal epithelial differentiation"],"prefix":"10.64898","author":[{"given":"No\u2019ad","family":"Shanas","sequence":"first","affiliation":[{"name":"Fels Cancer Institute for Personalized Medicine, Lewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aissata","family":"Balde","sequence":"additional","affiliation":[{"name":"Fels Cancer Institute for Personalized Medicine, Lewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annie D.","family":"Fuller","sequence":"additional","affiliation":[{"name":"Fels Cancer Institute for Personalized Medicine, Lewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jazmyne L.","family":"Jackson","sequence":"additional","affiliation":[{"name":"Fels Cancer Institute for Personalized Medicine, Lewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abigail J.","family":"Staub","sequence":"additional","affiliation":[{"name":"Fels Cancer Institute for Personalized Medicine, Lewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John M.","family":"Crespo","sequence":"additional","affiliation":[{"name":"Fels Cancer Institute for Personalized Medicine, Lewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William","family":"Nazario-Lugo","sequence":"additional","affiliation":[{"name":"Fels Cancer Institute for Personalized Medicine, Lewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Tufano","sequence":"additional","affiliation":[{"name":"Fels Cancer Institute for Personalized Medicine, Lewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas M.","family":"McDonald","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of Gastroenterology, Temple University Hospital, Philadelphia, PA 19140, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andres J.","family":"Klein-Szanto","sequence":"additional","affiliation":[{"name":"Histopathology Facility, Fox Chase Cancer Center, Philadelphia, PA 19111, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathy Q.","family":"Cai","sequence":"additional","affiliation":[{"name":"Histopathology Facility, Fox Chase Cancer Center, Philadelphia, PA 19111, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2943-9944","authenticated-orcid":false,"given":"Melanie","family":"Ruffner","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology, Hepatology, and Nutrition, The Children\u2019s Hospital of Philadelphia, Philadelphia, PA 19104, USA"},{"name":"Department of Pediatrics, The Children\u2019s Hospital of Philadelphia, Philadelphia, PA 19104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amanda B.","family":"Muir","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology, Hepatology, and Nutrition, The Children\u2019s Hospital of Philadelphia, Philadelphia, PA 19104, USA"},{"name":"Division of Allergy and Immunology, The Children\u2019s Hospital of Philadelphia, Philadelphia, PA 19104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zachary Wilmer","family":"Reichenbach","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of Gastroenterology, Temple University Hospital, Philadelphia, PA 19140, USA"},{"name":"Center for Substance Abuse Research (CSAR), Temple University Lewis Katz School of Medicine, Philadelphia, PA 19140, USA"},{"name":"Center for Microbiology and Immunology, Temple University Lewis Katz School of Medicine, Philadelphia, PA 19140, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7402-4935","authenticated-orcid":false,"given":"Kelly A.","family":"Whelan","sequence":"additional","affiliation":[{"name":"Fels Cancer Institute for Personalized Medicine, Lewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USA"},{"name":"Department of Cancer and Cellular Biology, Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026092209351072000_2026.09.17.752311v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2011.437"},{"key":"2026092209351072000_2026.09.17.752311v1.2","doi-asserted-by":"publisher","DOI":"10.1007\/BF00164174"},{"key":"2026092209351072000_2026.09.17.752311v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2016.11.042"},{"key":"2026092209351072000_2026.09.17.752311v1.4","doi-asserted-by":"publisher","DOI":"10.3390\/ijms241512181"},{"key":"2026092209351072000_2026.09.17.752311v1.5","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2020.580070"},{"key":"2026092209351072000_2026.09.17.752311v1.6","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.263403"},{"key":"2026092209351072000_2026.09.17.752311v1.7","doi-asserted-by":"publisher","DOI":"10.1126\/scisignal.2003638"},{"key":"2026092209351072000_2026.09.17.752311v1.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2021.108689"},{"key":"2026092209351072000_2026.09.17.752311v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.molmet.2025.102098"},{"key":"2026092209351072000_2026.09.17.752311v1.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcmgh.2026.101782"},{"key":"2026092209351072000_2026.09.17.752311v1.11","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2024.07.030"},{"key":"2026092209351072000_2026.09.17.752311v1.12","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2024-334561"},{"key":"2026092209351072000_2026.09.17.752311v1.13","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1000471"},{"key":"2026092209351072000_2026.09.17.752311v1.14","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.0903069"},{"key":"2026092209351072000_2026.09.17.752311v1.15","first-page":"729","article-title":"Telomerase induces immortalization of human esophageal keratinocytes without p16INK4a inactivation","volume":"1","year":"2003","journal-title":"Mol Cancer Res MCR"},{"key":"2026092209351072000_2026.09.17.752311v1.16","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2010.08.040"},{"key":"2026092209351072000_2026.09.17.752311v1.17","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0150968"},{"key":"2026092209351072000_2026.09.17.752311v1.18","doi-asserted-by":"publisher","DOI":"10.1152\/ajpendo.00457.2019"},{"key":"2026092209351072000_2026.09.17.752311v1.19","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrep.2021.100916"},{"key":"2026092209351072000_2026.09.17.752311v1.20","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0308970"},{"key":"2026092209351072000_2026.09.17.752311v1.21","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-29747-x"},{"key":"2026092209351072000_2026.09.17.752311v1.22","doi-asserted-by":"publisher","DOI":"10.1126\/science.1260419"},{"key":"2026092209351072000_2026.09.17.752311v1.23","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-019-1906-x"},{"key":"2026092209351072000_2026.09.17.752311v1.24","doi-asserted-by":"publisher","DOI":"10.1038\/mi.2013.90"},{"key":"2026092209351072000_2026.09.17.752311v1.25","doi-asserted-by":"publisher","DOI":"10.1172\/JCI26679"},{"key":"2026092209351072000_2026.09.17.752311v1.26","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2007.10.024"},{"key":"2026092209351072000_2026.09.17.752311v1.27","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2010.01.038"},{"key":"2026092209351072000_2026.09.17.752311v1.28","doi-asserted-by":"publisher","DOI":"10.2165\/11585450-000000000-00000"},{"key":"2026092209351072000_2026.09.17.752311v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcmgh.2024.02.018"},{"key":"2026092209351072000_2026.09.17.752311v1.30","doi-asserted-by":"publisher","DOI":"10.1038\/onc.2017.102"},{"key":"2026092209351072000_2026.09.17.752311v1.31","doi-asserted-by":"publisher","DOI":"10.1016\/j.exer.2018.06.003"},{"key":"2026092209351072000_2026.09.17.752311v1.32","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceca.2017.05.003"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.09.17.752311","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T16:35:17Z","timestamp":1790094917000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.09.17.752311"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,18]]},"references-count":32,"URL":"https:\/\/doi.org\/10.64898\/2026.09.17.752311","relation":{},"subject":[],"published":{"date-parts":[[2026,9,18]]},"subtype":"preprint"}}