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Potent inhibition of breast cancer by bis-indole-derived nuclear receptor 4A1 (NR4A1) antagonists

  • Preclinical study
  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Background

Nuclear receptor 4A1 (NR4A1) is overexpressed in mammary tumors, and the methylene-substituted bis-indole derivative 1,1-bis(3′-indolyl)-1-(p-hydroxyphenyl)methane (DIM-C-pPhOH) acts as an NR4A1 antagonist (inverse agonist) and inhibits NR4A1-regulated pro-oncogenic pathways/genes in breast and other cancer cells.

Methods

Buttressed analogs of DIM-C-pPhOH were synthesized by condensation of the substituted p-hydroxybenzaldehydes with indole. Breast cancer cell growth, survival, and migration assays were carried out by cell counting, Annexin V staining, and Boyden chamber assays, respectively. Changes in RNA and protein expression were determined by RT-PCR and western blots, respectively. Analysis of RNAseq results was carried out using Ingenuity Pathway Analysis, and in vivo potencies of NR4A1 antagonists were determined in athymic nude mice bearing MDA-MB-231 cells in an orthotopic model.

Results

Ingenuity Pathway analysis of common genes modulated by NR4A1 knockdown or treatment with DIM-C-pPhOH showed that changes in gene expression were consistent with the observed decreased functional responses, namely inhibition of growth and migration and increased apoptosis. DIM-C-pPhOH is rapidly metabolized and the effects and potencies of buttressed analogs of DIM-C-pPhOH which contain one or two substituents ortho to the hydroxyl groups were investigated using NR4A1-regulated gene/gene products as endpoints. The buttressed analogs were more potent than DIM-C-pPhOH in both in vitro assays and as inhibitors of mammary tumor growth. Moreover, using 1,1-bis(3′-indolyl)-1-(3-chloro-4-hydroxy-5-methoxyphenyl)methane (DIM-C-pPhOh-3-Cl-5-OCH3) significant tumor growth inhibition was observed at doses as low as 2 mg/kg/d which was at least an order of magnitude more potent than DIM-C-pPhOH.

Conclusions

These buttressed analogs represent a more potent set of second generation NR4A1 antagonists as inhibitors of breast cancer.

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Data availability

The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

C-DIMs:

Methylene-substituted DIMs

DIM-C-pPhOH:

1,1-bis(3′-indolyl)-1-(p-hydroxyphenyl)methane

DIM-C-pPhOh-3-Cl-5-OCH3 :

1,1-bis(3′-indolyl)-1-(3-chloro-4-hydroxy-5-methoxyphenyl)methane

DMSO:

Dimethyl sulfoxide

EGFR:

Epidermal growth factor receptor

NR4A1:

Nuclear receptor 4A1

SERMs:

Selective estrogen receptor modulators

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Acknowledgement

The financial assistance of the Sid Kyle Chair endowment, Texas AgriLife and the National Institutes of Health (P30-ES023512, S. Safe) is gratefully acknowledged.

Author information

Author notes

  1. Erik Hedrick

    Present address: Cleveland Clinic, Lerner Research Institute, Cleveland, OH, 44195, USA

  2. Yating Cheng

    Present address: MD Anderson Cancer Center, Houston, TX, 77030, USA

  3. Alexandra Lacey

    Present address: Shell Oil, Houston, TX, 77002, USA

Authors and Affiliations

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

    Erik Hedrick, Xi Li, Yating Cheng, Kumaravel Mohankumar, Mahsa Zarei & Stephen Safe

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  1. Erik Hedrick
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Contributions

SS conceived the study and wrote the manuscript. EH, XL, YC, AL and KM were involved with collection of data. EH, XL, YC, AL, KM and MZ carried out all data analysis. EH, XL, YC, AL, KM, MZ and SS were all involved in data interpretation, critical revisions of the manuscript, and approval of the final version.

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Correspondence to Stephen Safe.

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The Texas A&M University Institutional Animal Care and Use Committee reviewed and approved our animal treatment and use protocols.

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This article does not contain any studies with human participants performed by any of the authors.

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Hedrick, E., Li, X., Cheng, Y. et al. Potent inhibition of breast cancer by bis-indole-derived nuclear receptor 4A1 (NR4A1) antagonists. Breast Cancer Res Treat 177, 29–40 (2019). https://doi.org/10.1007/s10549-019-05279-9

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