Skip to main content

Advertisement

SpringerLink
Log in

Trastuzumab improves tumor perfusion and vascular delivery of cytotoxic therapy in a murine model of HER2+ breast cancer: preliminary results

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

Abstract

To employ in vivo imaging and histological techniques to identify and quantify vascular changes early in the course of treatment with trastuzumab in a murine model of HER2+ breast cancer. Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) was used to quantitatively characterize vessel perfusion/permeability (via the parameter K trans) and the extravascular extracellular volume fraction (v e ) in the BT474 mouse model of HER2+ breast cancer (N = 20) at baseline, day one, and day four following trastuzumab treatment (10 mg/kg). Additional cohorts of mice were used to quantify proliferation (Ki67), microvessel density (CD31), pericyte coverage (α-SMA) by immunohistochemistry (N = 44), and to quantify human VEGF-A expression (N = 29) throughout the course of therapy. Longitudinal assessment of combination doxorubicin ± trastuzumab (N = 42) tested the hypothesis that prior treatment with trastuzumab will increase the efficacy of subsequent doxorubicin therapy. Compared to control tumors, trastuzumab-treated tumors exhibited a significant increase in K trans (P = 0.035) on day four, indicating increased perfusion and/or vessel permeability and a simultaneous significant increase in v e (P = 0.01), indicating increased cell death. Immunohistochemical and ELISA analyses revealed that by day four the trastuzumab-treated tumors had a significant increase in vessel maturation index (i.e., the ratio of α-SMA to CD31 staining) compared to controls (P < 0.001) and a significant decrease in VEGF-A (P = 0.03). Additionally, trastuzumab dosing prior to doxorubicin improved the overall effectiveness of the therapies (P < 0.001). This study identifies and validates improved perfusion characteristics following trastuzumab therapy, resulting in an improvement in trastuzumab-doxorubicin combination therapy in a murine model of HER2+ breast cancer. This data suggests properties of vessel maturation. In particular, the use of DCE-MRI, a clinically available imaging method, following treatment with trastuzumab may provide an opportunity to optimize the scheduling and improve delivery of subsequent cytotoxic therapy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Abbreviations

α-SMA:

Alpha- smooth muscle actin

ANOVA:

Analysis of variance

BCA:

Bicinchoninic

DCE-MRI:

Dynamic contrast-enhanced magnetic resonance imaging

ELISA:

Enzyme-linked immunosorbent assay

Gd-DTPA:

Gadolinium-diethylenetriaminepentaacetic acid

HER2+:

Human epidermal growth factor receptor 2 positive

H&E:

Hematoxylin and eosin stain

MAPK:

Mitogen-activated protein kinase

MVD:

Microvessel density

NEX:

Number of excitations

PI3K/AKT:

Phosphoinositide 3-kinase/protein kinase B

ROC:

Receiver operating characteristic

ROI:

Region-of-interest

TE:

Echo time

TR:

Repetition time

VEGF:

Vascular endothelial growth factor

VMI:

Vessel maturation index

References

  1. Mitri Z, Constantine T, O’Regan R (2012) The HER2 receptor in breast cancer: pathophysiology, clinical use, and new advances in therapy. Chemother Res Pract 2012:743193

    PubMed Central  PubMed  Google Scholar 

  2. Dean-Colomb W, Esteva FJ (2008) Her2-positive breast cancer: herceptin and beyond. Eur J Cancer 44(18):2806–2812

    Article  CAS  PubMed  Google Scholar 

  3. Spector NL, Blackwell KL (2009) Understanding the mechanisms behind trastuzumab therapy for human epidermal growth factor receptor 2-positive breast cancer. J Clin Oncol 27(34):5838–5847

    Article  CAS  PubMed  Google Scholar 

  4. Nahta R, Esteva FJ (2006) HER2 therapy: molecular mechanisms of trastuzumab resistance. Breast Cancer Res (BCR) 8(6):215

    Article  Google Scholar 

  5. Cho HS, Mason K, Ramyar KX, Stanley AM, Gabelli SB, Denney DW Jr, Leahy DJ (2003) Structure of the extracellular region of HER2 alone and in complex with the Herceptin Fab. Nature 421(6924):756–760

    Article  CAS  PubMed  Google Scholar 

  6. Menard S, Pupa SM, Campiglio M, Tagliabue E (2003) Biologic and therapeutic role of HER2 in cancer. Oncogene 22(42):6570–6578

    Article  CAS  PubMed  Google Scholar 

  7. Vu T, Claret FX (2012) Trastuzumab: updated mechanisms of action and resistance in breast cancer. Front Oncol 2:62

    Article  PubMed Central  PubMed  Google Scholar 

  8. Klapper LN, Kirschbaum MH, Sela M, Yarden Y (2000) Biochemical and clinical implications of the ErbB/HER signaling network of growth factor receptors. Adv Cancer Res 77:25–79

    Article  CAS  PubMed  Google Scholar 

  9. Zhang A, Shen G, Zhao T, Zhang G, Liu J, Song L, Wei W, Bing L, Wu Z, Wu Q (2010) Augmented inhibition of angiogenesis by combination of HER2 antibody chA21 and trastuzumab in human ovarian carcinoma xenograft. J Ovarian Res 3:20

    Article  PubMed Central  PubMed  Google Scholar 

  10. Izumi Y, Xu L, di Tomaso E, Fukumura D, Jain RK (2002) Tumour biology: herceptin acts as an anti-angiogenic cocktail. Nature 416(6878):279–280

    Article  CAS  PubMed  Google Scholar 

  11. Klos KS, Zhou X, Lee S, Zhang L, Yang W, Nagata Y, Yu D (2003) Combined trastuzumab and paclitaxel treatment better inhibits ErbB-2-mediated angiogenesis in breast carcinoma through a more effective inhibition of Akt than either treatment alone. Cancer 98(7):1377–1385

    Article  CAS  PubMed  Google Scholar 

  12. Vogel CL, Cobleigh MA, Tripathy D, Gutheil JC, Harris LN, Fehrenbacher L, Slamon DJ, Murphy M, Novotny WF, Burchmore M, Shak S, Stewart SJ, Press M (2002) Efficacy and safety of trastuzumab as a single agent in first-line treatment of HER2-overexpressing metastatic breast cancer. J Clin Oncol 20(3):719–726

    Article  CAS  PubMed  Google Scholar 

  13. Hortobagyi GN (2005) Trastuzumab in the treatment of breast cancer. N Engl J Med 353(16):1734–1736

    Article  CAS  PubMed  Google Scholar 

  14. Jain RK, Carmeliet P (2012) SnapShot: tumor angiogenesis. Cell 149 (6):1408–1408 e1

  15. Jain RK (2013) Normalizing tumor microenvironment to treat cancer: bench to bedside to biomarkers. J Clin Oncol 31(17):2205–2218

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  16. Goel S, Wong AH, Jain RK (2012) Vascular normalization as a therapeutic strategy for malignant and nonmalignant disease. Cold Spring Harb Perspect Med 2(3):a006486

    Article  PubMed Central  PubMed  Google Scholar 

  17. Goel S, Duda DG, Xu L, Munn LL, Boucher Y, Fukumura D, Jain RK (2011) Normalization of the vasculature for treatment of cancer and other diseases. Physiol Rev 91(3):1071–1121

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Jain RK (2005) Normalization of tumor vasculature: an emerging concept in antiangiogenic therapy. Science 307(5706):58–62

    Article  CAS  PubMed  Google Scholar 

  19. Jain RK (2001) Normalizing tumor vasculature with anti-angiogenic therapy: a new paradigm for combination therapy. Nat Med 7(9):987–989. doi:10.1038/nm0901-987

    Article  CAS  PubMed  Google Scholar 

  20. Yankeelov TE, Gore JC (2009) Dynamic contrast enhanced magnetic resonance imaging in oncology: theory, data acquisition, analysis, and examples. Curr Med Imaging Rev 3(2):91–107

    Article  PubMed Central  PubMed  Google Scholar 

  21. Li X, Abramson RG, Arlinghaus LR, Kang H, Chakravarthy AB, Abramson VG, Farley J, Mayer IA, Kelley MC, Meszoely IM, Means-Powell J, Grau AM, Sanders M, Yankeelov TE (2015) Multiparametric magnetic resonance imaging for predicting pathological response after the first cycle of neoadjuvant chemotherapy in breast cancer. Invest Radiol 50(4):195–204

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Andersen EK, Hole KH, Lund KV, Sundfor K, Kristensen GB, Lyng H, Malinen E (2013) Pharmacokinetic parameters derived from dynamic contrast enhanced MRI of cervical cancers predict chemoradiotherapy outcome. Radiother Oncol 107(1):117–122

    Article  PubMed  Google Scholar 

  23. Hylton NM, Blume JD, Bernreuter WK, Pisano ED, Rosen MA, Morris EA, Weatherall PT, Lehman CD, Newstead GM, Polin S, Marques HS, Esserman LJ, Schnall MD, Team AT, Investigators IST (2012) Locally advanced breast cancer: MR imaging for prediction of response to neoadjuvant chemotherapy–results from ACRIN 6657/I-SPY TRIAL. Radiology 263(3):663–672

    Article  PubMed Central  PubMed  Google Scholar 

  24. Hirashima Y, Yamada Y, Tateishi U, Kato K, Miyake M, Horita Y, Akiyoshi K, Takashima A, Okita N, Takahari D, Nakajima T, Hamaguchi T, Shimada Y, Shirao K (2012) Pharmacokinetic parameters from 3-Tesla DCE-MRI as surrogate biomarkers of antitumor effects of bevacizumab plus FOLFIRI in colorectal cancer with liver metastasis. Int J Cancer 130(10):2359–2365

    Article  CAS  PubMed  Google Scholar 

  25. O’Connor JP, Jackson A, Parker GJ, Jayson GC (2007) DCE-MRI biomarkers in the clinical evaluation of antiangiogenic and vascular disrupting agents. Br J Cancer 96(2):189–195

    Article  PubMed Central  PubMed  Google Scholar 

  26. Barnes SL, Whisenant JG, Loveless ME, Ayers GD, Yankeelov TE (2013) Assessing the reproducibility of dynamic contrast enhanced magnetic resonance imaging in a murine model of breast cancer. Magn Reson Med 69(6):1721–1734

    Article  PubMed Central  PubMed  Google Scholar 

  27. Barnes SL, Whisenant JG, Loveless ME, Yankeelov TE (2012) Practical dynamic contrast enhanced MRI in small animal models of cancer: data acquisition, data analysis, and interpretation. Pharmaceutics 4(3):442–478

    Article  PubMed Central  PubMed  Google Scholar 

  28. Wu X, Jeong EK, Emerson L, Hoffman J, Parker DL, Lu ZR (2010) Noninvasive evaluation of antiangiogenic effect in a mouse tumor model by DCE-MRI with Gd-DTPA cystamine copolymers. Mol Pharm 7(1):41–48

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  29. Baselga J, Norton L, Albanell J, Kim YM, Mendelsohn J (1998) Recombinant humanized anti-HER2 antibody (Herceptin) enhances the antitumor activity of paclitaxel and doxorubicin against HER2/neu overexpressing human breast cancer xenografts. Cancer Res 58(13):2825–2831

    CAS  PubMed  Google Scholar 

  30. Lewis GD, Figari I, Fendly B, Wong WL, Carter P, Gorman C, Shepard HM (1993) Differential responses of human tumor cell lines to anti-p185HER2 monoclonal antibodies. Cancer Immunol Immunother (CII) 37(4):255–263

    Article  CAS  Google Scholar 

  31. Whisenant JG, McIntyre JO, Peterson TE, Kang H, Sanchez V, Manning HC, Arteaga CL, Yankeelov TE (2014) Utility of [F]FLT-PET to assess treatment response in trastuzumab-resistant and trastuzumab-sensitive HER2-overexpressing human breast cancer xenografts. Mol Imaging Biol (MIB) 17(1):119–128

    Article  Google Scholar 

  32. Neve RM, Chin K, Fridlyand J, Yeh J, Baehner FL, Fevr T, Clark L, Bayani N, Coppe JP, Tong F, Speed T, Spellman PT, DeVries S, Lapuk A, Wang NJ, Kuo WL, Stilwell JL, Pinkel D, Albertson DG, Waldman FM, McCormick F, Dickson RB, Johnson MD, Lippman M, Ethier S, Gazdar A, Gray JW (2006) A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. Cancer Cell 10(6):515–527

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  33. Whisenant JG, Sorace AG, McIntyre JO, Kang H, Sanchez V, Loveless ME, Yankeelov TE (2014) Evaluating treatment response using DW-MRI and DCE-MRI in trastuzumab responsive and resistant HER2-overexpressing human breast cancer xenografts. Transl Oncol 7(6):768–779

    Article  PubMed Central  PubMed  Google Scholar 

  34. Hurley J, Doliny P, Reis I, Silva O, Gomez-Fernandez C, Velez P, Pauletti G, Powell JE, Pegram MD, Slamon DJ (2006) Docetaxel, cisplatin, and trastuzumab as primary systemic therapy for human epidermal growth factor receptor 2-positive locally advanced breast cancer. J Clin Oncol 24(12):1831–1838

    Article  CAS  PubMed  Google Scholar 

  35. Romond EH, Perez EA, Bryant J, Suman VJ, Geyer CE Jr, Davidson NE, Tan-Chiu E, Martino S, Paik S, Kaufman PA, Swain SM, Pisansky TM, Fehrenbacher L, Kutteh LA, Vogel VG, Visscher DW, Yothers G, Jenkins RB, Brown AM, Dakhil SR, Mamounas EP, Lingle WL, Klein PM, Ingle JN, Wolmark N (2005) Trastuzumab plus adjuvant chemotherapy for operable HER2-positive breast cancer. N Engl J Med 353(16):1673–1684

    Article  CAS  PubMed  Google Scholar 

  36. Tofts PS, Brix G, Buckley DL, Evelhoch JL, Henderson E, Knopp MV, Larsson HB, Lee TY, Mayr NA, Parker GJ, Port RE, Taylor J, Weisskoff RM (1999) Estimating kinetic parameters from dynamic contrast-enhanced T(1)-weighted MRI of a diffusible tracer: standardized quantities and symbols. J Magn Reson Imaging (JMRI) 10(3):223–232

    Article  CAS  Google Scholar 

  37. Daldrup H, Shames DM, Wendland M, Okuhata Y, Link TM, Rosenau W, Lu Y, Brasch RC (1998) Correlation of dynamic contrast-enhanced magnetic resonance imaging with histologic tumor grade: comparison of macromolecular and small-molecular contrast media. Pediatr Radiol 28(2):67–78

    Article  CAS  PubMed  Google Scholar 

  38. de Lussanet QG, Beets-Tan RG, Backes WH, van der Schaft DW, van Engelshoven JM, Mayo KH, Griffioen AW (2004) Dynamic contrast-enhanced magnetic resonance imaging at 1.5 Tesla with gadopentetate dimeglumine to assess the angiostatic effects of anginex in mice. Eur J Cancer 40(8):1262–1268

    Article  PubMed  Google Scholar 

  39. Vangestel C, Van de Wiele C, Van Damme N, Staelens S, Pauwels P, Reutelingsperger CP, Peeters M (2011) (99)mTc-(CO)(3) His-annexin A5 micro-SPECT demonstrates increased cell death by irinotecan during the vascular normalization window caused by bevacizumab. J Nucl Med 52(11):1786–1794. doi:10.2967/jnumed.111.092650

    Article  CAS  PubMed  Google Scholar 

  40. Ahmed S, Sami A, Xiang J (2015) HER2-directed therapy: current treatment options for HER2-positive breast cancer. Breast Cancer 22(2):101–116

    Article  PubMed  Google Scholar 

  41. Heyerdahl H, Roe K, Brevik EM, Dahle J (2013) Modifications in dynamic contrast-enhanced magnetic resonance imaging parameters after alpha-particle-emitting (2)(2)(7)Th-trastuzumab therapy of HER2-expressing ovarian cancer xenografts. Int J Radiat Oncol Biol Phys 87(1):153–159

    Article  CAS  PubMed  Google Scholar 

  42. McCormack DR, Walsh AJ, Sit W, Arteaga CL, Chen J, Cook RS, Skala MC (2014) In vivo hyperspectral imaging of microvessel response to trastuzumab treatment in breast cancer xenografts. Biomed Opt Express 5(7):2247–2261

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  43. Le XF, Mao W, Lu C, Thornton A, Heymach JV, Sood AK, Bast RC Jr (2008) Specific blockade of VEGF and HER2 pathways results in greater growth inhibition of breast cancer xenografts that overexpress HER2. Cell Cycle 7(23):3747–3758

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank the National Cancer Institute for support through R01CA138599, P50CA098131, P30CA68485, R25CA092043, 5T32CA093240, and U01CA174706. We thank the Kleberg Foundation for the generous support of Vanderbilt's biomedical imaging program. The authors thank Dr. Carlos Arteaga for his guidance on the combination therapy portions of this project. Additionally, we would also like to thank Dr. Jin Chen, Dr. Shan Wang, Dr. Melissa Skala, and Ms. Amy Shah for their helpful conversations.

Authors’ contributions

AS and JW carried out DCE-MRI studies, AS and VS carried out histology studies. AS and AH performed the ELISA study. AS carried out the combination therapeutic studies. AS completed the statistical analysis. OM, CQ, AS, AH, JW, and TY were involved in the conception and design of the study. All authors read and approved the final manuscript.

Author information

Authors and Affiliations

  1. Department of Radiology and Radiological Sciences, Vanderbilt University, Nashville, TN, USA

    Anna G. Sorace, Jennifer G. Whisenant, J. Oliver McIntyre & Thomas E. Yankeelov

  2. Vanderbilt Institute of Imaging Science, Vanderbilt University Medical Center, AA-1105 Medical Center North, 1161 21st Ave South, Nashville, TN, 37232-2310, USA

    Anna G. Sorace, Jennifer G. Whisenant, J. Oliver McIntyre & Thomas E. Yankeelov

  3. Division of Neuroimaging Research, Barrow Neurological Institute, Dignity Health, St. Joseph’s Hospital and Medical Center, Phoenix, AZ, 85013, USA

    C. Chad Quarles

  4. Department of Cancer Biology, Vanderbilt University, Nashville, TN, USA

    Ariella B. Hanker, J. Oliver McIntyre & Thomas E. Yankeelov

  5. Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, USA

    Thomas E. Yankeelov

  6. Department of Hematology Oncology, Vanderbilt University, Nashville, TN, USA

    Violeta M. Sanchez

  7. Vanderbilt Ingram Cancer Center, Vanderbilt University, Nashville, TN, USA

    Thomas E. Yankeelov

  8. Department of Physics and Astronomy, Vanderbilt University, Nashville, TN, USA

    Thomas E. Yankeelov

Authors
  1. Anna G. Sorace
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. Chad Quarles
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jennifer G. Whisenant
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ariella B. Hanker
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J. Oliver McIntyre
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Violeta M. Sanchez
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Thomas E. Yankeelov
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anna G. Sorace.

Ethics declarations

Conflicts of interest

The authors have no competing interest to declare in relation to this manuscript.

Additional information

The following authors’ affliations have changed to The University of Texas at Austin: AGS (anna.sorace@austin.utexas.edu) and TEY (thomas.yankeelov@utexas.edu).

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sorace, A.G., Quarles, C.C., Whisenant, J.G. et al. Trastuzumab improves tumor perfusion and vascular delivery of cytotoxic therapy in a murine model of HER2+ breast cancer: preliminary results. Breast Cancer Res Treat 155, 273–284 (2016). https://doi.org/10.1007/s10549-016-3680-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10549-016-3680-8

Keywords

Search

Navigation