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Tissue factor expression in ovarian cancer: implications for immunotherapy with hI-con1, a factor VII-IgGFc chimeric protein targeting tissue factor

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Abstract

We evaluated the expression of tissue factor (TF) in ovarian cancer (EOC) and the potential of hI-con1, an antibody-like molecule targeting TF, as a novel form of therapy against chemotherapy-resistant ovarian disease. We studied the expression of TF in 88 EOC by immunohistochemistry (IHC) and real-time-PCR (qRT-PCR) and the levels of membrane-bound-complement-regulatory-proteins CD46, CD55 and CD59 in primary EOC cell lines by flow-cytometry. Sensitivity to hI-con1-dependent-cell-mediated-cytotoxicity (IDCC), complement-dependent-cell-cytotoxicity and inhibition of IDCC by γ-immunoglobulin were evaluated in 5-h 51chromium-release-assays. Cytoplasmic and/or membrane TF expression was observed in 24 out of 25 (96%) of the EOC samples tested by IHC, but not in normal ovarian-tissue. EOC with clear cell histology significantly overexpress TF when compared to serous, endometrioid, or undifferentiated tumors by qRT-PCR. With a single exception, all primary EOC that overexpressed TF demonstrated high levels of CD46, CD55 and CD59 and regardless of their histology or resistance to chemotherapy, were highly sensitive to IDCC. The effect of complement and physiologic doses of γ-immunoglobulin on IDCC in ovarian cancer cell lines overexpressing TF was tumor specific and related to the overexpression of CD59 on tumor cells. Small-interfering-RNA-mediated knockdown of CD59 expression in ovarian tumors significantly increased hI-con1-mediated cytotoxic activity in vitro. Finally, low doses of interleukin-2 further increased the cytotoxic effect induced by hI-con1 (P < 0.01). hI-con1 molecule induces strong cytotoxicity against primary chemotherapy-resistant ovarian cancer cell lines overexpressing TF and may represent a novel therapeutic agent for the treatment of ovarian tumors refractory to standard treatment modalities.

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Abbreviations

TF:

Tissue factor

fVII:

Factor VII

hI-con1:

Human immuno-conjugate molecule

IDCC:

hI-con1-dependent cell-mediated cytotoxicity

FBS:

Fetal bovine serum

IHC:

Immunohistochemistry

mAb:

Monoclonal antibody

NK cells:

Natural killer cells

PBL:

Peripheral blood lymphocytes

qRT-PCR:

Quantitative real-time-polymerase chain reaction

CC:

Clear cell carcinoma

OSPC:

Ovarian serous papillary carcinoma

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Acknowledgments

Supported in part by grants from NIH R01 CA122728-01A2 to AS, and grants 501/A3/3 and 0027557 from the Italian Institute of Health (ISS) to AS. This investigation was also supported by NIH Research Grant CA-16359 from the National Cancer Institute. The Authors thank Dr. William Konigsberg, Dr. Alan Garen and Dr. Zhiwei Hu for initiating the collaboration with CJL on Icon immunotherapy of human gynecologic malignancies and Iconic Therapeutics Inc. for providing hI-con1 protein free of charge for our studies.

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Authors and Affiliations

  1. Department of Obstetrics, Gynecology & Reproductive Sciences, Yale University School of Medicine, 305 LSOG, 333 Cedar Street, P.O. Box 208063, New Haven, CT, 06520-8063, USA

    Emiliano Cocco, Joyce Varughese, Stefania Bellone, Ken-Yu Lin, Marta Bellone, Paola Todeschini, Dan-Arin Silasi, Masoud Azodi, Peter E. Schwartz, Thomas J. Rutherford, Luisa Carrara, Charles J. Lockwood & Alessandro D. Santin

  2. Pathology, Yale University School of Medicine, New Haven, CT, USA

    Natalia Buza

  3. Division of Gynecologic Oncology, University of Brescia, Brescia, Italy

    Renata Tassi & Sergio Pecorelli

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  1. Emiliano Cocco
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  2. Joyce Varughese
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  3. Natalia Buza
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  4. Stefania Bellone
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  6. Marta Bellone
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  7. Paola Todeschini
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  8. Dan-Arin Silasi
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  9. Masoud Azodi
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  10. Peter E. Schwartz
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  11. Thomas J. Rutherford
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  14. Sergio Pecorelli
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  15. Charles J. Lockwood
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  16. Alessandro D. Santin
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Correspondence to Alessandro D. Santin.

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Cocco, E., Varughese, J., Buza, N. et al. Tissue factor expression in ovarian cancer: implications for immunotherapy with hI-con1, a factor VII-IgGFc chimeric protein targeting tissue factor. Clin Exp Metastasis 28, 689–700 (2011). https://doi.org/10.1007/s10585-011-9401-0

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  • DOI: https://doi.org/10.1007/s10585-011-9401-0

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