Ablation of the Renal Tubular Gluconeogenic Enzyme PCK1 Drives AKI-to-CKD Transition by Negatively Regulating the TGF-β/Smad3 Signaling Pathway

Ablation of the Renal Tubular Gluconeogenic Enzyme PCK1 Drives AKI-to-CKD Transition by Negatively Regulating the TGF-β/Smad3 Signaling Pathway | 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 Research Article Ablation of the Renal Tubular Gluconeogenic Enzyme PCK1 Drives AKI-to-CKD Transition by Negatively Regulating the TGF-β/Smad3 Signaling Pathway Jingxuan Gan, Mengfan Li, Haihua Deng, Qiutong Liu, Zhixiao Chen, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9202714/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 5 You are reading this latest preprint version Abstract The transition from acute kidney injury (AKI) to chronic kidney disease (CKD) is characterized by progressive renal fibrosis, yet the metabolic drivers of this maladaptive repair remain inadequately defined. In this study, we investigated the role of phosphoenolpyruvate carboxykinase 1 (PCK1), the rate-limiting enzyme of gluconeogenesis, in the AKI-to-CKD transition. Analysis of human single-cell RNA sequencing (scRNA-seq) datasets revealed a profound downregulation of PCK1 in injured proximal tubule cells, which correlated with the upregulation of pro-fibrotic markers. Using a longitudinal cisplatin-induced mouse model, we demonstrated that the loss of PCK1 precedes significant collagen deposition and acts as a primary driver of fibrogenesis. Mechanistically, PCK1 deficiency triggers a metabolic shift that facilitates the activation of the TGF-β/Smad3 signaling pathway. Our functional assays established that PCK1 serves as an endogenous antagonist of Smad3, its depletion promotes Smad3 phosphorylation and nuclear translocation, leading to the loss of tubular epithelial identity. Significantly, the pharmacological or genetic restoration of PCK1 effectively blunted Smad3 activation and attenuated renal fibrosis in vivo and in vitro. These findings identify PCK1 as a critical metabolic regulator that maintains tubular homeostasis and suggest that PCK1-targeted metabolic restoration may be promising therapeutic strategies for arresting the progression of chronic kidney disease following acute injury. AKI-to-CKD transition PCK1 renal fibrosis TGF-β/Smad signaling proximal tubular cells Full Text Supplementary Files SupplementaryFiguresandTable.pdf Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Major Revision 23 Apr, 2026 Reviewers agreed at journal 30 Mar, 2026 Reviewers invited by journal 29 Mar, 2026 Editor assigned by journal 26 Mar, 2026 First submitted to journal 26 Mar, 2026 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. 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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-9202714","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":614156426,"identity":"5118cbe8-1fd6-4bae-8812-b0bde5879c24","order_by":0,"name":"Jingxuan Gan","email":"","orcid":"","institution":"Guangzhou University of Traditional Chinese Medicine: Guangzhou University of Chinese Medicine","correspondingAuthor":false,"prefix":"","firstName":"Jingxuan","middleName":"","lastName":"Gan","suffix":""},{"id":614156427,"identity":"eb6c79d7-4d0b-47ce-b723-aaf2b44bc2df","order_by":1,"name":"Mengfan 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