title: Identification of approved and investigational drugs that inhibit hypoxia-inducible factor-1 signaling.
abstract: One of the requirements for tumor development is blood supply, most often driven by hypoxia-induced angiogenesis. Hypoxia induces the stabilization of hypoxia-inducible factor-1 alpha (HIF-1α), which induces expression of an angiogenic factor, vascular endothelial growth factor (VEGF). The purpose of this study is to validate a new screening platform combined with orthogonal assays to rapidly identify HIF-1 inhibitors and to evaluate the effectiveness of approved drugs on modulating HIF-1 signaling. We generated an endogenous HIF-1α-NanoLuc luciferase reporter allele in the human HCT116 colon cancer cell line using genome editing and screened a panel of small interfering RNAs (siRNAs) to 960 druggable targets and approximately 2,500 drugs on a quantitative high-throughput screening (qHTS) platform. Selected compounds were further investigated with secondary assays to confirm their anti-HIF activity and to study their mode of action. The qHTS assay identified over 300 drugs that inhibited HIF-1α-NanoLuc expression. The siRNA screening results supported the effectiveness of several target-specific inhibitors. Moreover, the identified HIF-1 inhibitors, such as mycophenolate mofetil, niclosamide, and trametinib, were able to suppress cancer cell proliferation and angiogenesis. Our study indicates that blocking the mitogen-activated protein kinase (MAPK) and phosphoinositol 3-kinase (PI3K) pathways effectively inhibits hypoxia-induced HIF-1α accumulation and HIF-1α transactivation and that proteasome inhibitors induce accumulation and decrease transcriptional activity of HIF-1α. These findings underline the importance of developing a battery of robust assay platforms and confirmation studies that focus on endogenous protein targets so that only relevant and reliable data will be taken into pre-clinical and clinical studies.
authors.affiliation: Division of Pre-Clinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Chia-Wen
authors.initials: CW
authors.lastname: Hsu
authors.affiliation: Division of Pre-Clinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Ruili
authors.initials: R
authors.lastname: Huang
authors.affiliation: Division of Pre-Clinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Thai
authors.initials: T
authors.lastname: Khuc
authors.affiliation: Division of Pre-Clinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
authors.forename: David
authors.initials: D
authors.lastname: Shou
authors.affiliation: Horizon Discovery Ltd., Waterbeach, Cambridge, UK.
authors.forename: Joshua
authors.initials: J
authors.lastname: Bullock
authors.affiliation: Horizon Discovery Ltd., Waterbeach, Cambridge, UK.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Grooby
authors.affiliation: Horizon Discovery Ltd., Waterbeach, Cambridge, UK.
authors.forename: Sue
authors.initials: S
authors.lastname: Griffin
authors.affiliation: American Type Culture Collection, Gaithersburg, MD, USA.
authors.forename: Chaozhong
authors.initials: C
authors.lastname: Zou
authors.affiliation: Horizon Discovery Ltd., Waterbeach, Cambridge, UK.
authors.forename: Annette
authors.initials: A
authors.lastname: Little
authors.affiliation: Horizon Discovery Ltd., Waterbeach, Cambridge, UK.
authors.forename: Holly
authors.initials: H
authors.lastname: Astley
authors.affiliation: Division of Pre-Clinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Menghang
authors.initials: M
authors.lastname: Xia
mesh_terms: D015687:Cell Hypoxia; D049109:Cell Proliferation; D003110:Colonic Neoplasms / Q000098:blood supply / Q000188:drug therapy* / Q000235:genetics* / Q000473:pathology; D015507:Drugs, Investigational / Q000494:pharmacology*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000037:antagonists & inhibitors* / Q000235:genetics; D008156:Luciferases / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D036103:Nanotechnology; D009389:Neovascularization, Pathologic; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured
chemical_list: D015507:Drugs, Investigational; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D008156:Luciferases; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D020928:Mitogen-Activated Protein Kinases
keywords: cancer; drug; genome editing; high-throughput screening; hypoxia inducible factor