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Host genetic modifiers of nonproductive angiogenesis inhibit breast cancer

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

Abstract

Purpose

Multiple aspects of the tumor microenvironment (TME) impact breast cancer, yet the genetic modifiers of the TME are largely unknown, including those that modify tumor vascular formation and function.

Methods

To discover host TME modifiers, we developed a system called the Consomic/Congenic Xenograft Model (CXM). In CXM, human breast cancer cells are orthotopically implanted into genetically engineered consomic xenograft host strains that are derived from two parental strains with different susceptibilities to breast cancer. Because the genetic backgrounds of the xenograft host strains differ, whereas the inoculated tumor cells are the same, any phenotypic variation is due to TME-specific modifier(s) on the substituted chromosome (consomic) or subchromosomal region (congenic). Here, we assessed TME modifiers of growth, angiogenesis, and vascular function of tumors implanted in the SSIL2Rγ and SS.BN3IL2Rγ CXM strains.

Results

Breast cancer xenografts implanted in SS.BN3IL2Rγ (consomic) had significant tumor growth inhibition compared with SSIL2Rγ (parental control), despite a paradoxical increase in the density of blood vessels in the SS.BN3IL2Rγ tumors. We hypothesized that decreased growth of SS.BN3IL2Rγ tumors might be due to nonproductive angiogenesis. To test this possibility, SSIL2Rγ and SS.BN3IL2Rγ tumor vascular function was examined by dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI), micro-computed tomography (micro-CT), and ex vivo analysis of primary blood endothelial cells, all of which revealed altered vascular function in SS.BN3IL2Rγ tumors compared with SSIL2Rγ. Gene expression analysis also showed a dysregulated vascular signaling network in SS.BN3IL2Rγ tumors, among which DLL4 was differentially expressed and co-localized to a host TME modifier locus (Chr3: 95–131 Mb) that was identified by congenic mapping.

Conclusions

Collectively, these data suggest that host genetic modifier(s) on RNO3 induce nonproductive angiogenesis that inhibits tumor growth through the DLL4 pathway.

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Abbreviations

TME:

Tumor microenvironment

CXM:

Consomic Xenograft Model

DCE-MRI:

Dynamic contrast-enhanced magnetic resonance imaging

SSRS:

Species-specific RNAseq

PBST:

Phosphate-buffered saline plus Tween-20

RNO3:

Rat chromosome 3

TNBC:

Triple-negative breast cancer

micro-CT:

Micro-computed tomography

EC:

Endothelial cell

DMEM:

Dulbecco’s modifier Eagle’s medium

MFP:

Mammary fat pad

RARE:

Rapid acquisition rapid echo

IAUC:

Initial area under the curve

ROI:

Region of interest

FDR:

False discovery rate

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Acknowledgements

We thank M. Tschannen, R. Schilling, E. Schneider, A. Zappa, and Y. Liu for excellent technical support and the Center for Imaging Research in the Medical College of Wisconsin Department of Radiology, and the Biomedical Imaging Shared Resource supported by the MCW Cancer Center.

Funding

This work was supported by a seed grant from the Wisconsin Breast Cancer Showhouse and the MCW Cancer Center, the Mary Kay Foundation (Grant No. 024.16), and the NCI (R01CA193343) to M.J.F. Support was also received from the National Center for Research Resources, the National Center for Advancing Translational Sciences, and the Office of the Director of the NIH via the Clinical & Translational Science Institute (#8KL2TR000056), the Wisconsin Breast Cancer Showhouse and the MCW Cancer Center, the Rosenberg Translational Research Award, and an institutional research Grant (#86-004-26) from the American Cancer Society to C.B.

Author information

Authors and Affiliations

  1. Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, WI, USA

    Michael J. Flister, Shirng-Wern Tsaih, Alexander Stoddard, Cody Plasterer, Angela Lemke, Dana Murphy, Aron M. Geurts & Melinda R. Dwinell

  2. Cancer Center, Medical College of Wisconsin, Milwaukee, WI, USA

    Michael J. Flister, Shirng-Wern Tsaih, Cody Plasterer, Jaidip Jagtap, Abdul K. Parchur, Gayatri Sharma, Angela Lemke, Dana Murphy, Mona Al-Gizawiy, Michael Straza, Melinda R. Dwinell, Carmen Bergom, Peter S. LaViolette & Amit Joshi

  3. Department of Physiology, Medical College of Wisconsin, 8701 Watertown Plank Rd., Milwaukee, WI, 53226, USA

    Michael J. Flister, Shirng-Wern Tsaih, Cody Plasterer, Anthony R. Prisco, Angela Lemke, Dana Murphy, Aron M. Geurts, Melinda R. Dwinell & Andrew S. Greene

  4. Department of Radiology, Medical College of Wisconsin, Milwaukee, WI, USA

    Jaidip Jagtap, Abdul K. Parchur, Gayatri Sharma, Mona Al-Gizawiy, Peter S. LaViolette & Amit Joshi

  5. Department of Radiation Oncology, Medical College of Wisconsin, Milwaukee, WI, USA

    Michael Straza & Carmen Bergom

  6. Simmons Cancer Institute, Southern Illinois University School of Medicine, Springfield, IL, USA

    Sophia Ran

  7. Department of Medical Microbiology, Immunology, and Cell Biology, Southern Illinois University School of Medicine, Springfield, IL, USA

    Sophia Ran

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  1. Michael J. Flister
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Corresponding author

Correspondence to Michael J. Flister.

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The authors have no conflict of interests to declare.

Ethical approval

All applicable international, national, and institutional guidelines for the care and use of animals were followed. The Institutional Animal Care and Use Committee (IACUC) of the Medical College of Wisconsin approved all animal studies. All procedures involving animals were conducted in accordance with the National Institutes of Health guidelines concerning the use and care of experimental animals.

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Flister, M.J., Tsaih, SW., Stoddard, A. et al. Host genetic modifiers of nonproductive angiogenesis inhibit breast cancer. Breast Cancer Res Treat 165, 53–64 (2017). https://doi.org/10.1007/s10549-017-4311-8

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