The transcription factor HHEX maintains glucocorticoid levels and protects adrenals from androgen-induced lipid depletion

The transcription factor HHEX maintains glucocorticoid levels and protects adrenals from androgen-induced lipid depletion | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article The transcription factor HHEX maintains glucocorticoid levels and protects adrenals from androgen-induced lipid depletion Typhanie Dumontet, Kaitlin J. Basham, Micah C. Foster, Emma Silverman, and 11 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6248794/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 11 Jan, 2026 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Glucocorticoid-producing cells of the adrenal cortex ( i.e. zona fasciculata, zF) constitute the critical effectors of the hypothalamic-pituitary-adrenal axis, mediating the mammalian stress response. With glucocorticoids being essential for life, it is not surprising that zF dysfunction perturbs multiple organs that participate in optimizing cardiometabolic fitness. The zF forms a dynamic and heterogenous cell population endowed with the capacity to remodel through the engagement of both proliferative and differentiation programs that enable the adrenal to adapt and respond to diverse stressors. However, the mechanisms that sustain such differential responsiveness remain poorly understood. In this study, we resolved the transcriptome of the steroidogenic lineage by scRNA-seq using Sf1-Cre; Rosa mT/mG reporter mice. We identified HHEX, a homeodomain protein, as the most enriched transcription factor in glucocorticoid-producing cells. We developed new genetic mouse models to demonstrate that HHEX deletion causes glucocorticoid insufficiency in male animals. Molecularly, we demonstrated that HHEX is an androgen receptor (AR) target gene, shaping the sexual dimorphism of the adrenal gland by repressing the female transcriptional program at puberty, while also maintaining zF cholesterol ester content by protecting lipid droplets from androgen-induced-lipophagy. Moreover, our study revealed that, in both sexes, HHEX is crucial for maintaining the identity of the innermost adrenocortical cell subpopulation. Specifically, loss of HHEX impairs the expression of Abcb1b (P-glycoprotein/MDR1), an efflux pump regulating steroid export and cellular levels of xenobiotics. Together, these data demonstrate that HHEX serves as a multi-functional regulator of post-natal adrenal maturation that is potentiated by androgens. Biological sciences/Cell biology Biological sciences/Physiology Steroidogenic lineage PRH zona fasciculata corticosteroid cholesterol endocrine autophagy lipophagy lipid droplet testosterone sexual dimorphism ACTH stress response HPA axis Full Text Additional Declarations Yes there is potential Competing Interest. G.D.H. Founder and Board of Directors – Sling Therapeutics, Advisor – Orphagen Pharmaceuticals Supplementary Files 20250317SupplementaryTables.xlsx Cite Share Download PDF Status: Published Journal Publication published 11 Jan, 2026 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6248794","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":442447364,"identity":"a771cafa-89dd-48fd-a1f0-999dd5756691","order_by":0,"name":"Typhanie Dumontet","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA20lEQVRIiWNgGAWjYFCCBAjFD8QSQMwDJXEDHogWAwbJBpK1GByAaGEgqMWePcd0w8cdf+SMrx0+eIOxbZsM/+wGxgdv2/DYwvPG7ObMMwbGZrfTki0Y227zSNw5wGw4F58WiRyz27xtBonbbueYSYC0GEgksEnzEqGlfvPs/G8wLey/idGSYCCdwwa3hRmvljPPym7ObDM2nHE7zdgi4RzQLzcSmyXnnMOthb09eduNj21y8vyzkx/e+FB2255/RvLBD2/KcGtBBQlgkrGBWPWjYBSMglEwCnAAAJYsTS4iQ1txAAAAAElFTkSuQmCC","orcid":"","institution":"Training Program in Organogenesis, Center for Cell Plasticity and Organ Design, University of Michigan, Ann Arbor, Michigan, USA","correspondingAuthor":true,"prefix":"","firstName":"Typhanie","middleName":"","lastName":"Dumontet","suffix":""},{"id":442447365,"identity":"f02670e4-ac66-4382-b431-c8d2c93142bf","order_by":1,"name":"Kaitlin J. 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With glucocorticoids being essential for life, it is not surprising that zF dysfunction perturbs multiple organs that participate in optimizing cardiometabolic fitness. The zF forms a dynamic and heterogenous cell population endowed with the capacity to remodel through the engagement of both proliferative and differentiation programs that enable the adrenal to adapt and respond to diverse stressors. However, the mechanisms that sustain such differential responsiveness remain poorly understood. In this study, we resolved the transcriptome of the steroidogenic lineage by scRNA-seq using \u003ci\u003eSf1-Cre; Rosa\u003csup\u003emT/mG\u003c/sup\u003e\u003c/i\u003e reporter mice. We identified HHEX, a homeodomain protein, as the most enriched transcription factor in glucocorticoid-producing cells. We developed new genetic mouse models to demonstrate that HHEX deletion causes glucocorticoid insufficiency in male animals. Molecularly, we demonstrated that HHEX is an androgen receptor (AR) target gene, shaping the sexual dimorphism of the adrenal gland by repressing the female transcriptional program at puberty, while also maintaining zF cholesterol ester content by protecting lipid droplets from androgen-induced-lipophagy. Moreover, our study revealed that, in both sexes, HHEX is crucial for maintaining the identity of the innermost adrenocortical cell subpopulation. Specifically, loss of HHEX impairs the expression of \u003ci\u003eAbcb1b\u003c/i\u003e (P-glycoprotein/MDR1), an efflux pump regulating steroid export and cellular levels of xenobiotics. Together, these data demonstrate that HHEX serves as a multi-functional regulator of post-natal adrenal maturation that is potentiated by androgens.","manuscriptTitle":"The transcription factor HHEX maintains glucocorticoid levels and protects adrenals from androgen-induced lipid depletion","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-15 02:06:45","doi":"10.21203/rs.3.rs-6248794/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-communications","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"NCOMMS","sideBox":"Learn more about [Nature Communications](http://www.nature.com/ncomms/)","snPcode":"","submissionUrl":"https://mts-ncomms.nature.com/","title":"Nature Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Communications","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"7bb684ce-3b90-4558-bad9-aaf5cf2670c9","owner":[],"postedDate":"April 15th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":47159476,"name":"Biological sciences/Cell biology"},{"id":47159477,"name":"Biological sciences/Physiology"}],"tags":[],"updatedAt":"2026-02-11T08:11:04+00:00","versionOfRecord":{"articleIdentity":"rs-6248794","link":"https://doi.org/10.1038/s41467-025-68257-4","journal":{"identity":"nature-communications","isVorOnly":false,"title":"Nature Communications"},"publishedOn":"2026-01-11 05:00:00","publishedOnDateReadable":"January 11th, 2026"},"versionCreatedAt":"2025-04-15 02:06:45","video":"","vorDoi":"10.1038/s41467-025-68257-4","vorDoiUrl":"https://doi.org/10.1038/s41467-025-68257-4","workflowStages":[]},"version":"v1","identity":"rs-6248794","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6248794","identity":"rs-6248794","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}