Graduation Year

2006

Document Type

Dissertation

Degree

Ph.D.

Degree Granting Department

Molecular Medicine

Major Professor

Richard Heller, Ph.D.

Keywords

In vivo electroporation, Long-term survival, Tumor regression, Vpr, Anti-cancer agent

Abstract

Novel therapies and delivery methods directed against malignancies such as melanoma, and particularly metastatic melanoma, are needed. The HIV-1 accessory protein Vpr (viral protein R) has previously been demonstrated to induce G2 cell cycle arrest as well as in vitro growth inhibition/killing of numerous tumor cell lines. In vivo electroporation has been utilized as an effective delivery method for pharmacologic agents as well as DNA plasmids that express "therapeutic" proteins and has been targeted to various tissues including malignant tumors. In this study, we assessed the ability of electroporation-mediated delivery of Vpr plasmid (pVpr) to induce growth attenuation or complete tumor regression in C57BL/6 mice with subcutaneous B16.F10 melanoma lesions.

To assess the administration of intratumoral delivery of pVpr with in vivo electroporation, a range of Vpr plasmid dosages, electroporation parameters, and treatment days were evaluated in a subcutaneous B16 murine melanoma model. pVpr was injected directly into the tumors. Immediately following the injection, the subcutaneous tumors were electroporated. Treatment with 25 microgram or 100 microgram of pVpr plus electroporation on days 0 and 4 resulted in complete tumor regressions with long-term survival in 14.3% and 7.1% of the mice, respectively. In order to optimize the treatment regimen, B16 tumors were treated on days 0, 2, and 4 with 100 microgram pVpr plus electroporation which resulted in 50% of the mice with complete tumor regressions and long-term survival. Additional investigations revealed intratumoral Vpr expression and demonstrated that apoptosis was the mechanism by which Vpr caused tumor regression in vivo.

This study confirmed that treatment with 100 microgram of pVpr plus electroporation led to durable complete regressions in established murine melanoma lesions. The pVpr plus electroporation treatment regimen has induced complete regressions in mice as well as resistance to tumor challenge in some of the animals. This is the first comprehensive study demonstrating the ability of Vpr, when delivered as a DNA expression plasmid with in vivo electroporation, to induce complete tumor regressions coupled with long- term survival of mice in a highly aggressive and metastatic solid tumor model.

Download

Share

COinS