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Anti-tumor and anti-metastasis efficacy of E6201, a MEK1 inhibitor, in preclinical models of triple-negative breast cancer

  • Jangsoon Lee
  • Bora Lim
  • Troy Pearson
  • Kuicheon Choi
  • Jon A. Fuson
  • Chandra Bartholomeusz
  • Linda J. Paradiso
  • Thomas Myers
  • Debu Tripathy
  • Naoto T. UenoEmail author
Preclinical study
  • 266 Downloads

Abstract

Purpose

Triple-negative breast cancer (TNBC) lacks the receptor targets estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2, and thus, it does not respond to receptor-targeted treatments. TNBC has higher recurrence, metastasis, and mortality rates than other subtypes of breast cancer. Mounting data suggest that the MAPK (also known as RAS-RAF-MEK-ERK) pathway is an important therapeutic target in TNBC.

Methods

To evaluate anti-tumor and anti-metastasis efficacy of E6201, we used cell proliferation assay, soft agar assay, cell cycle assay, Annexin V staining assay, immunoblotting analysis, immunohistochemistry, migration assay, invasion assay, mammary fat pad xenograft, and experimental and spontaneous metastasis xenograft models. We also evaluated the anti-tumor efficacy of E6201 plus CDK4/6 inhibitor, mTOR inhibitor, or ATR inhibitor.

Results

E6201 inhibited TNBC cell colony formation, migration, and invasion in a dose-dependent manner. E6201 induced G1 cell cycle arrest and apoptosis. E6201 inhibited TNBC xenograft growth and inhibited TNBC lung metastasis and improved mouse survival in experimental metastasis and spontaneous metastasis assays. Immunohistochemical staining demonstrated that E6201 decreased the metastatic burden in the lung and decreased phosphorylated ERK expression in a dose-dependent manner. Combination of E6201 with CDK4/6 inhibitor or mTOR inhibitor enhanced E6201’s in vitro anti-tumor efficacy.

Conclusion

These results indicate that E6201 exhibits anti-tumor efficacy against TNBC in vitro and anti-metastasis efficacy against TNBC in vivo. These results provide a rationale for further clinical development of E6201 as a MAPK-pathway-targeted therapy for TNBC.

Keywords

E6201 MEK inhibitor MAPK pathway Triple-negative breast cancer Metastasis 

Notes

Acknowledgements

We thank Sunita Patterson and Stephanie Deming of the Department of Scientific Publications at MD Anderson Cancer Center for editing the manuscript.

Funding

This work was supported by the Morgan Welch Inflammatory Breast Cancer Research Program, the State of Texas Rare and Aggressive Breast Cancer Research Program, the National Institutes of Health/National Cancer Institute (Grant CA123318), MD Anderson’s Cancer Center Support Grant (P30CA016672, used the Characterized Cell Line Core Facility and Flow Cytometry and Cellular Imaging Facility), and Spirita Oncology, LLC.

Compliance with ethical standards

Conflict of interest

Naoto T. Ueno has a research agreement with Spirita Oncology, LLC. Linda J. Paradiso and Thomas Myers are employees of Spirita Oncology, LLC. All other authors declare no potential conflicts of interest.

Research involving in animal rights

Animal studies and procedures were approved by The University of Texas MD Anderson Cancer Center Animal Care and Use Committee. Protocol #: 00000968-RN02 (approval date 5/1/2015, expiration date 4/22/2021).

Research involving in human rights

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

Supplementary material

10549_2019_5166_MOESM1_ESM.pptx (1.8 mb)
Supplemental Figure 1. Anti-proliferation effect of MEK inhibitors in TNBC cell lines. TNBC cells were treated with selumetinib, pimasertib, or trametinib for 5 days, and viability was measured by using CellTiter-Blue and sulforhodamine-B assays. Data shown are representative of three experiments with similar results. Each point represents the mean of three independent experiments; error bars indicate standard deviation. Supplemental Figure 2. FLT3 inhibition does not affect TNBC cell proliferation, migration, or invasion. a Western blotting of cells treated with E6201 for the indicated times. b Proliferation assay. TNBC cell lines were treated with quizartinib for 5 days, and viability was measured by using CellTiter-Blue. c and d Migration and invasion assays. TNBC cells (1×105/well) were added into trans-wells with or without quizartinib for 6 hr (migration, c) or 24 hr (invasion, d). Migration and invasion were evaluated by using ImageJ software. Data shown are representative of three experiments with similar results; error bars indicate standard deviation. Supplementary material 1 (PPTX 1826 KB)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Jangsoon Lee
    • 1
    • 3
  • Bora Lim
    • 1
    • 3
  • Troy Pearson
    • 1
  • Kuicheon Choi
    • 1
  • Jon A. Fuson
    • 1
  • Chandra Bartholomeusz
    • 1
    • 3
  • Linda J. Paradiso
    • 2
  • Thomas Myers
    • 2
  • Debu Tripathy
    • 1
  • Naoto T. Ueno
    • 1
    • 3
    Email author
  1. 1.Section of Translational Breast Cancer Research, Morgan Welch Inflammatory Breast Cancer Research Program and Clinic, Department of Breast Medical OncologyThe University of Texas MD Anderson Cancer CenterHoustonUSA
  2. 2.Spirita Oncology, LLCNatickUSA
  3. 3.Morgan Welch Inflammatory Breast Cancer Research Program and Clinic, Department of Breast Medical OncologyThe University of Texas MD Anderson Cancer CenterHoustonUSA

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