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AAV-P125A-endostatin and paclitaxel treatment increases endoreduplication in endothelial cells and inhibits metastasis of breast cancer

Gene Therapy volume 18, pages 145–154 (2011)Cite this article

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Abstract

Endostatin potentiates the antimitotic effects of paclitaxel (taxol) on endothelial cells (ECs). P125A-endostatin and taxol-treated ECs showed multipolar spindles and nuclear lobulation, leading to mitotic catastrophe and cell death. Induction of nuclear abnormalities was found to be dependent on β-catenin levels as wnt-mediated overexpression of β-catenin reversed the changes in nuclear morphology. These results prompted us to investigate whether antiangiogenic gene therapy and paclitaxel chemotherapy can synergistically inhibit angiogenesis and tumor growth. We first determined the effect of combination treatment in a transgenic mouse model of breast cancer. Intramuscular injection of recombinant adeno-associated virus type-2 virus induced sustained expression of P125A-endostatin. In vivo studies showed that combination therapy inhibited mammary cancer growth, delayed the onset of multifocal mammary adenocarcinomas, decreased tumor angiogenesis and increased survival in treated mice. In a second model, female athymic mice were orthotopically transplanted with a metastatic human breast cancer cell line. Antiangiogenic gene therapy in combination with paclitaxel inhibited tumor angiogenesis and lung/lymph-node metastasis in this model. These studies demonstrate cooperation between endostatin gene therapy and chemotherapy to inhibit tumor initiation, growth and metastasis.

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Abbreviations

AAV-2:

recombinant adeno-associated virus type-2

VEGF:

vascular endothelial growth factor

P125A-endostatin:

human endostatin in which proline 125 was substituted by alanine

AAV-P125A-endostatin:

recombinant adeno-associated virus expressing P125A-endostatin

AAV-LacZ:

control recombinant adeno-associated virus expressing LacZ

EC:

endothelial cells

Ad HEK293 cells:

adherent human embryonic kidney cells

BrdU:

5-bromo-2′deoxyuridine

HUVEC:

human umbilical vein endothelial cells

v.p.:

viral particles

MOI:

multiplicity of infection

DAPI:

4′-6-diamidino-2-phenylindole

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Acknowledgements

This work was supported, in part, by grants from the NIH CA-114340 and Shirley Ann Sparboe Endowment for Women's Cancer Research. We acknowledge Mr G Sedgwick and Mr J Oja of Biomedical Imaging and Processing Laboratory for helping with the confocal images. We also thank Ms Julia Nguyen for her technical support.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology and Women's Health, University of Minnesota Medical School, Minneapolis, MN, USA

    I V Subramanian, R Ghebre & S Ramakrishnan

  2. Department of Pharmacology, University of Minnesota Medical School, Minneapolis, MN, USA

    I V Subramanian, G Ghosh, H P Joshi, Y Jing & S Ramakrishnan

  3. Department of Hematology, Oncology and Transplantation, University of Minnesota Medical School, Minneapolis, MN, USA

    S Devineni

  4. Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis, MN, USA

    A M Truskinovsky

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  1. I V Subramanian
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Correspondence to S Ramakrishnan.

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Subramanian, I., Devineni, S., Ghebre, R. et al. AAV-P125A-endostatin and paclitaxel treatment increases endoreduplication in endothelial cells and inhibits metastasis of breast cancer. Gene Ther 18, 145–154 (2011). https://doi.org/10.1038/gt.2010.118

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