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Nuclear receptor 4A2 (NR4A2) is a druggable target for glioblastomas

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Abstract

Introduction

The orphan nuclear receptor 4A2 (NR4A2) has been extensively characterized in subcellular regions of the brain and is necessary for the function of dopaminergic neurons. The NR4A2 ligand, 1,1-bis (31-indoly1)-1-(p-chlorophenyl)methane (DIM-C-pPhCl) inhibits markers of neuroinflammation and degeneration in mouse models and in this study we investigated expression and function of NR4A2 in glioblastoma (GBM).

Methods

Established and patient-derived cell lines were used as models and the expression and functions of NR4A2 were determined by western blots and NR4A2 gene silencing by antisense oligonucleotides respectively. Effects of NR4A2 knockdown and DIM-C-pPhCl on cell growth, induction of apoptosis (Annexin V Staining) and migration/invasion (Boyden chamber and spheroid invasion assay) and transactivation of NR4A2-regulated reporter genes were determined. Tumor growth was investigated in athymic nude mice bearing U87-MG cells as xenografts.

Results

NR4A2 knockdown and DIM-C-pPhCl inhibited GBM cell and tumor growth, induced apoptosis and inhibited migration and invasion of GBM cells. DIM-C-pPhCl and related analogs also inhibited NR4A2-regulated transactivation (luciferase activity) confirming that DIM-C-pPhCl acts as an NR4A2 antagonist and blocks NR4A2-dependent pro-oncogenic responses in GBM.

Conclusion

We demonstrate for the first time that NR4A2 is pro-oncogenic in GBM and thus a potential druggable target for patients with tumors expressing this receptor. Moreover, our bis-indole-derived NR4A2 antagonists represent a novel class of anti-cancer agents with potential future clinical applications for treating GBM.

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Acknowledgements

This work was supported by the National Institutes of Health [P30-ES023512 (SS), R01-ES025713 (SS), R01-CA202697 (SS), and T32-ES026568 (KK)], Texas A&M AgriLife Research (SS), the Sid Kyle Chair Endowment (SS), and the Karmanos Cancer Institute (SM).

Funding

This research received federal grant and University funding as indicated under funding.

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Authors and Affiliations

  1. Department of Veterinary Physiology and Pharmacology, Texas A&M University, College Station, TX, 77843, USA

    Keshav Karki, Xi Li, Un-Ho Jin, Kumaravel Mohankumar, Mahsa Zarei & Stephen Safe

  2. Department of Surgery, Virginia Tech University, Roanoke, VA, 24016, USA

    Sharon K. Michelhaugh & Sandeep Mittal

  3. Department of Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, CO, 80523, USA

    Ronald Tjalkens

  4. Department of Veterinary Physiology & Pharmacology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, 4466 TAMU, College Station, TX, 77843-4466, USA

    Stephen Safe

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  1. Keshav Karki
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  2. Xi Li
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  4. Kumaravel Mohankumar
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  9. Stephen Safe
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Contributions

Study conception and design: S.K.M., S.M., S.H.S. Analysis and interpretation of data: All authors. Experimentation: K.K., S.K.M. Generation of Data: K.K., X.L., U.J., K.M., M.Z., S.K.M. Drafting of manuscript: S.K.M., S.M., S.H.S. Final approval of article: All authors.

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Correspondence to Stephen Safe.

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Karki, K., Li, X., Jin, UH. et al. Nuclear receptor 4A2 (NR4A2) is a druggable target for glioblastomas. J Neurooncol 146, 25–39 (2020). https://doi.org/10.1007/s11060-019-03349-y

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