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GDNF induces RET–SRC–HER2-dependent growth in trastuzumab-sensitive but SRC-independent growth in resistant breast tumor cells

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Abstract

We investigated the role of glial cell line-derived neurotrophic factor (GDNF) in compensating trastuzumab (TZMB)-induced apoptosis in HER2+ breast cancer (BC) cells using xenograft tumors. We generated BC xenografts in nude mice using samples from three patients selected based on their HER2 status and response to TZMB therapy. TZMB treatment resulted in shrinkage of the HER2+ TZMB-sensitive xenograft tumor but not the HER2− or HER2+ TZMB-resistant ones. GDNF neutralized TZMB activity and induced growth in all tumors. Three distinct cell lines were derived from these tumors and named, respectively, TZMB-sensitive (TSTC), HER2− (HNTC), and TZMB-resistant (TRTC). Over 50% of TRTC but 1% of TSTC cells expressed CD44, whereas 84% of TSTC were CD24+ compared to only 1% of TRTC, despite comparable levels of HER2 detected in both. TZMB induced profound morphological changes toward apoptosis in TSTC but not in TRTC or HNTC. However, GDNF significantly compensated TZMB-mediated TSTC cell loss and promoted growth by 37 and 50%, respectively, in TSTC and TRTC. Inhibition of SRC by Saracatinib (SARC) blocked GDNF function and accelerated TZMB-mediated cell death in TSTC, but GDNF continued promoting TRTC growth. These changes paralleled with expression levels of the key molecules involved in growth and apoptosis. Collectively, we found in our xenograft samples that firstly SRC mediates GDNF pro-survival functions by bridging RET–HER2 crosstalk in TZMB-responsive BC tumors. Secondly, SARC–TZMB interactions can synergistically eradicate such tumor cells; and thirdly, GDNF can support antibody resistance by acting independent from SRC in tumors with poor HER2 response to TZMB therapy.

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Acknowledgement

We thank those patients who contributed to this study by donating their tumor samples. We also appreciate collaborations made by staff in Imam Khomeini Hospital in Tehran for generation of xenograft models of tumors. This work was supported by a Grant (II-502) from NIGEB.

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Author notes

  1. Babak Behnam

    Present address: NIH Undiagnosed Diseases Program, National Human Genome Research Institute (NHGRI), National Institute of Health (NIH), Bethesda, MD, USA

Authors and Affiliations

  1. Department of Stem Cells and Regenerative Medicine, Faculty of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Pazhoohesh Blvd, Tehran-Karaj HWY Kilometer 15, PO BOX 14965/161, Tehran, Iran

    Mossa Gardaneh & Sahar Shojaei

  2. Department of Surgery, Tehran University of Medical Sciences, Tehran, Iran

    Ahmad Kaviani

  3. Department of Medical Genetics and Molecular Biology, Faculty of Medicine, Iran University of Medical Sciences (IUMS), Tehran, Iran

    Babak Behnam

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  1. Mossa Gardaneh
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Correspondence to Mossa Gardaneh.

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Ethical approval

Informed consent was obtained from all individual participants included in the study. All procedures performed in studies involving human participants were in accordance with the ethical standards of our Institutional Ethics Committee, Ministry of Health in Iran and the 1964 Helsinki declaration and its later amendments. Also, all applicable international, national guidelines, the ARRIVE guidelines [29] besides those set by our Institutional Ethical Committee for the care and use of animals were followed.

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Gardaneh, M., Shojaei, S., Kaviani, A. et al. GDNF induces RET–SRC–HER2-dependent growth in trastuzumab-sensitive but SRC-independent growth in resistant breast tumor cells. Breast Cancer Res Treat 162, 231–241 (2017). https://doi.org/10.1007/s10549-016-4078-3

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  • DOI: https://doi.org/10.1007/s10549-016-4078-3

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