Development of Targeted Sindbis Virus Vectors for Potential Application to Breast Cancer Therapy
Annual summary 10 Aug 1999-9 Aug 2000
JOHNS HOPKINS UNIV BALTIMORE MD SCHOOLOF MEDICINE
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The future progress of cancer gene therapy relies on the development of efficient and safe vectors that can deliver therapeutic genes specifically to tumor cells. Using a replication-competent viral vector targeted to tumor cells may be the most efficient way of specifically killing a large number of malignant cells. We intend to develop Sindbis virus SV, an aiphavirus, into a targeted replication-competent viral vector for breast cancer gene therapy. Since SV kills by apoptosis, specific destruction of tumor cells will occur if the virus is targeted to breast cancer cells. To target SV to breast cancer cells, the putative receptor-binding domains of the SV E2 glycoprotein was replaced with the ligand, heregulin, or with an NGR-containing peptide motif that binds to the CD13 receptor expressed on tumor associated endothelial cells. We have demonstrated that a heregulin-containing SV preferentially kills a breast cancer cell line that expresses the appropriate epidermal growth factor receptors. Analysis of protein expression via metabolic labeling, protein immunoblotting or indirect immunofluorescence microscopy revealed synthesis of the viral structural proteins and the heregulin-containing E2 glycoprotein. SV vectors expressing the green fluorescent protein are being used to study the replication and spread of heregulin- and NGR-containing SV vectors in various cell lines.
- Genetic Engineering and Molecular Biology
- Medicine and Medical Research