Research Papers:

Identification of approved and investigational drugs that inhibit hypoxia-inducible factor-1 signaling

Chia-Wen Hsu, Ruili Huang, Thai Khuc, David Shou, Joshua Bullock, Suzanne Grooby, Sue Griffin, Chaozhong Zou, Annette Little, Holly Astley and Menghang Xia _

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Oncotarget. 2016; 7:8172-8183. https://doi.org/10.18632/oncotarget.6995

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Chia-Wen Hsu1, Ruili Huang1, Thai Khuc1, David Shou1, Joshua Bullock2, Suzanne Grooby2, Sue Griffin2, Chaozhong Zou3, Annette Little2, Holly Astley2, Menghang Xia1

1Division of Pre-Clinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA

2Horizon Discovery Ltd., Waterbeach, Cambridge, UK

3American Type Culture Collection, Gaithersburg, MD, USA

Correspondence to:

Menghang Xia, e-mail: [email protected]

Keywords: hypoxia inducible factor, cancer, genome editing, drug, high-throughput screening

Received: September 10, 2015     Accepted: January 05, 2016     Published: January 23, 2016


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.

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