{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T21:19:25Z","timestamp":1792790365168,"version":"4.5.0"},"posted":{"date-parts":[[2026,9,23]]},"group-title":"Developmental Biology","reference-count":32,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:00:00Z","timestamp":1790121600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"National Institute of General Medical Sciences","award":["1R16GM145545"],"award-info":[{"award-number":["1R16GM145545"]}],"id":[{"id":"https:\/\/ror.org\/04q48ey07","id-type":"ROR","asserted-by":"crossref"}]},{"name":"National Institute on Alcohol Abuse and Alcoholism","award":["U54 AA019765"],"award-info":[{"award-number":["U54 AA019765"]}],"id":[{"id":"https:\/\/ror.org\/02jzrsm59","id-type":"ROR","asserted-by":"crossref"}]},{"name":"National Institute on Minority Health and Health Disparities","award":["U54 MD012392"],"award-info":[{"award-number":["U54 MD012392"]}],"id":[{"id":"https:\/\/ror.org\/0493hgw16","id-type":"ROR","asserted-by":"crossref"}]},{"award":["1R16GM145545"],"award-info":[{"award-number":["1R16GM145545"]}],"id":[{"id":"https:\/\/ror.org\/04q48ey07","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,23]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  In utero alcohol exposure (IUE) can alter mammary development and increase mammary tumor susceptibility in experimental models. However, most previous studies have relied on carcinogen-induced or genetically tumor-prone models, leaving it unresolved whether prenatal alcohol exposure produces mammary developmental and molecular changes before the introduction of an oncogenic challenge. Here, we addressed this question using non-tumorigenic C57BL\/6 mice. Pregnant dams received a Lieber\u2013DeCarli liquid diet containing 1.7% (v\/v) ethanol or an isocaloric control diet from gestational day 11 to 19, and mammary glands from female offspring were examined during puberty (postnatal day 40, PND40) and young adulthood (PND70). At PND40, IUE enhanced mammary ductal morphogenesis, increased terminal end bud numbers, and elevated epithelial proliferation. These developmental alterations persisted at PND70, with increased lateral branching and proliferative activity. IUE also altered mammary epithelial composition, expanding the basal\/myoepithelial compartment and the CD24\n                  <jats:sup>high<\/jats:sup>\n                  CD49f\n                  <jats:sup>high<\/jats:sup>\n                  mammary repopulating unit\/stem cell-enriched population. Mammary epithelial cells from IUE-exposed offspring exhibited increased colony-forming cells (CFCs), mammosphere formation, and 3D Matrigel growth, indicating enhanced stem\/progenitor-associated activity. At the molecular level, IUE increased ER\u03b1 expression and phosphorylation together with Cyclin D1, c-Myc, E2F1, and Bcl-2, and enhanced AKT, ERK, and STAT3 phosphorylation, IL-6 expression, and SOX2 at PND40. Importantly, major components of this signaling phenotype remained altered at PND70, including ER\u03b1-associated signaling, AKT\/ERK activation, and IL-6\/STAT3 signaling, whereas the pubertal increase in SOX2 was not sustained. Together, these findings demonstrate that prenatal alcohol exposure induces profound and persistent histomorphological, cellular, and signaling alterations in the mammary gland in the absence of a carcinogenic challenge or genetically introduced oncogenic driver. These results identify intrinsic developmental reprogramming of normal mammary tissue as a potential basis through which prenatal alcohol exposure may modify subsequent responses to tumor-promoting or other environmental challenges.\n                <\/jats:p>","DOI":"10.64898\/2026.09.22.753591","type":"posted-content","created":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T16:05:19Z","timestamp":1790179519000},"source":"Crossref","is-referenced-by-count":0,"title":["In Utero Alcohol Exposure Reprograms Mammary Gland Development and Stem\/Progenitor Cell Activity in a Non-Tumorigenic Mouse Model"],"prefix":"10.64898","author":[{"given":"Zhikun","family":"Ma","sequence":"first","affiliation":[{"name":"Julius L. Chambers Biomedical\/Biotechnology Research Institute, North Carolina Research Campus, North Carolina Central University, 500 Laureate Way, Kannapolis, NC, 28081, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amanda B","family":"Parris","sequence":"additional","affiliation":[{"name":"Julius L. Chambers Biomedical\/Biotechnology Research Institute, North Carolina Research Campus, North Carolina Central University, 500 Laureate Way, Kannapolis, NC, 28081, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evelyn I","family":"Espinoza","sequence":"additional","affiliation":[{"name":"Julius L. Chambers Biomedical\/Biotechnology Research Institute, North Carolina Research Campus, North Carolina Central University, 500 Laureate Way, Kannapolis, NC, 28081, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohe","family":"Yang","sequence":"additional","affiliation":[{"name":"Julius L. Chambers Biomedical\/Biotechnology Research Institute, North Carolina Research Campus, North Carolina Central University, 500 Laureate Way, Kannapolis, NC, 28081, USA"},{"name":"Department of Biological and Biomedical Sciences, North Carolina Research Campus, North Carolina Central University, 500 Laureate Way, Kannapolis, NC, 28081, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026100310150578000_2026.09.22.753591v1.1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-pharmtox-010611-134659"},{"key":"2026100310150578000_2026.09.22.753591v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.alcohol.2005.04.005"},{"key":"2026100310150578000_2026.09.22.753591v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/nrc3220"},{"key":"2026100310150578000_2026.09.22.753591v1.4","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1849509"},{"key":"2026100310150578000_2026.09.22.753591v1.5","doi-asserted-by":"publisher","DOI":"10.1530\/ERC-15-0263"},{"key":"2026100310150578000_2026.09.22.753591v1.6","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00040.2018"},{"key":"2026100310150578000_2026.09.22.753591v1.7","doi-asserted-by":"publisher","DOI":"10.1289\/ehp.1002864"},{"issue":"1","key":"2026100310150578000_2026.09.22.753591v1.8","doi-asserted-by":"crossref","first-page":"660","DOI":"10.1038\/s42003-021-02201-2","article-title":"Mammary cell gene expression atlas links epithelial cell remodeling events to breast carcinogenesis","volume":"4","year":"2021","journal-title":"Communications biology"},{"issue":"Suppl 3","key":"2026100310150578000_2026.09.22.753591v1.9","first-page":"S3","article-title":"Alcohol consumption and breast cancer risk","volume":"11","year":"2009","journal-title":"Breast cancer research: BCR"},{"issue":"1","key":"2026100310150578000_2026.09.22.753591v1.10","doi-asserted-by":"crossref","first-page":"37","DOI":"10.35946\/arcr.v35.1.05","article-title":"In utero alcohol exposure, epigenetic changes, and their consequences","volume":"35","year":"2013","journal-title":"Alcohol research : current reviews"},{"key":"2026100310150578000_2026.09.22.753591v1.11","doi-asserted-by":"crossref","unstructured":"Mead EA , Sarkar DK : Fetal alcohol spectrum disorders and their transmission through genetic and epigenetic mechanisms. 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