Synergistic Inhibition of PI3K and HSP90 Enhanced Antitumorigenic Efficacy in Adrenocortical Carcinoma

Synergistic Inhibition of PI3K and HSP90 Enhanced Antitumorigenic Efficacy in Adrenocortical Carcinoma | 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 Synergistic Inhibition of PI3K and HSP90 Enhanced Antitumorigenic Efficacy in Adrenocortical Carcinoma Prachi Mishra, Brieann Sobieski, Dipranjan Laha, Steven D. Forsythe, and 10 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7761877/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Adrenocortical cancer (ACC) is a rare and aggressive malignancy with poor survival due to a lack of effective treatments; therefore, it is important to identify therapies to be readily studied in clinical trials. Quantitative high-throughput drug combination screening identified potent synergy between phosphatidylinositol-3-kinase (PI3K) inhibitor, PIK75 and heat shock protein 90 (HSP90) inhibitors, Ganetespib (STA9090), HSP990, or Luminespib (NVP-AUY922). Preclinical in-vitro and in-vivo studies were performed to validate the synergistic efficacy of the most effective HSP90 inhibitor and PI3K inhibitor combination in ACC cell lines, human ACC xenografts and patient-derived organoids (PDOs). Combination of PIK75 and STA9090, synergistically inhibited cell proliferation (monolayer and 3-dimensional), cell migration/invasion and epithelial-to-mesenchymal transition with decreased phosphorylated proteins in PI3K/mTOR signaling pathway. Due to the unavailabilityof PIK75 for clinical trial, another PI3K inhibitor, BGT226, which was clinically available and demonstrated a comparable synergistic efficacy with STA9090, was validated in the ACC cell lines. RNA sequencing analysis and phenotypic studies revealed that the BGT226-STA9090 combination induced autophagy-related cell death in ACC cells, unlike the PIK75-STA9090 combination which induced caspase-dependent apoptosis and G2/M cell cycle arrest. Further antitumor efficacy was confirmed by the BGT226-STA9090 combination in human ACC xenograft model and five PDOs with different pathogenic mutations. Conclusively, the combinations of PI3K and HSP90 inhibitors were highly effective in preclinical studies, warranting a clinical trial in patients with advanced ACC. Biological sciences/Cancer Biological sciences/Cell biology Biological sciences/Drug discovery Health sciences/Oncology Adrenocortical cancer Heat shock protein 90 PI3K combination therapy quantitative high-throughput drug screening Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementaryinformationMaterialsandMethods.pdf westernfullscansformanuscript.pdf Supplementaryfigures.pdf Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 22 Nov, 2025 Reviews received at journal 20 Nov, 2025 Reviews received at journal 11 Nov, 2025 Reviewers agreed at journal 27 Oct, 2025 Reviewers agreed at journal 23 Oct, 2025 Reviewers agreed at journal 22 Oct, 2025 Reviewers invited by journal 21 Oct, 2025 Editor assigned by journal 20 Oct, 2025 Submission checks completed at journal 20 Oct, 2025 First submitted to journal 01 Oct, 2025 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. 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Quantitative high-throughput drug combination screening identified potent synergy between phosphatidylinositol-3-kinase (PI3K) inhibitor, PIK75 and heat shock protein 90 (HSP90) inhibitors, Ganetespib (STA9090), HSP990, or Luminespib (NVP-AUY922). Preclinical \u003cem\u003ein-vitro\u003c/em\u003e and \u003cem\u003ein-vivo\u003c/em\u003estudies were performed to validate the synergistic efficacy of the most effective HSP90 inhibitor and PI3K inhibitor combination in\u003cstrong\u003e \u003c/strong\u003eACC cell lines, human ACC xenografts and patient-derived organoids (PDOs). Combination of\u003cstrong\u003e \u003c/strong\u003ePIK75 and STA9090, synergistically inhibited cell proliferation (monolayer and 3-dimensional), cell migration/invasion and epithelial-to-mesenchymal transition with decreased phosphorylated proteins in PI3K/mTOR signaling pathway. 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