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Development of a Targeted Adenoviral Vector Expressing HSV-TK for use in Breast Cancer Gene Therapy and Analysis through Positron Emission Tomography Open Access

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Other title
Subject/Keyword
Gene Therapy
Adenovirus
Breast Cancer
PET
HSV-TK
Positron Emission Tomography
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
DeSilva, Alan D
Supervisor and department
Hitt, Mary (Oncology)
Mercer, John (Oncology)
Examining committee member and department
Mittal, Suresh (Veterinary Medicine)
Wiebe, Leonard (Oncology)
Pasdar, Manijeh (Cell Biology)
Underhill, Alan (Oncology)
Hitt, Mary (Oncology)
Mercer, John (Oncology)
Department
Department of Oncology
Specialization
Experimental Oncology
Date accepted
2012-08-27T14:09:51Z
Graduation date
2012-11
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
While adenoviral (Ad) vectors are the most commonly used gene delivery vector for human gene therapy, improvements must be made to increase Ad specificity to tumors if they are to be used for cancer gene therapy. Our goal is to target adenoviral vectors to breast cancer (BrCa) cells to induce cell killing while reducing toxicity to non-tumor cells. We have generated a non-replicating BrCa-targeted adenoviral vector that utilizes a mammary-specific promoter (MPE2) to drive expression of a therapeutic gene, herpes simplex virus-1 thymidine kinase (HSV-TK). Cells expressing HSV-TK are sensitive to the prodrug ganciclovir (GCV). Thus, AdMPE2TK can induce BrCa-specific cell death when administered in combination with GCV. The expression of HSV-TK also allows cells to be visualized using positron emission tomography (PET) using radiolabeled PET substrates 3’-[18F]fluoro-3’-deoxythymidine ([18F]FLT) and 9-(4-[18F]fluoro-3-hydroxymethylbutyl)-guanine ([18F]FHBG). We first characterized the immunocompetent MTHJ murine breast cancer model as having high biological significance to human breast cancer and reveal its ability to uptake standard PET tracers. We then showed the BrCa specificity of the AdMPE2TK vector to kill BrCa cells in vitro in comparison to the non-specific AdCMVTK vector utilizing the cytomegalovirus promoter. Neither vector was shown to induce in vivo tumor regression, however the AdCMVTK vector caused liver toxicity in immunocompetent mice. In contrast, the AdMPE2TK vector did not induce any measurable toxicity, highlighting its specificity and potential for cancer gene therapy. Finally, using MTHJ murine tumor cells, in vitro cell uptake experiments revealed the ability of AdTK vectors to induce an increase in accumulation of [18F]FHBG. In vivo PET imaging was then used to evaluate the accumulation of [18F]FLT and [18F]FHBG in established MTHJ tumors injected with AdTK vectors, however no increase in accumulation was observed. This thesis outlines the first preclinical evaluation of the BrCa-specific MPE2 promoter to target HSV-TK for cancer gene therapy. We illustrate the ability of AdMPE2TK to induce a therapeutic effect in combination with GCV. In addition, this project represents the first imaging studies of an adenoviral vector utilizing the MPE2 promoter. This research outlines the potential of the MPE2 promoter and illustrates its application in BrCa gene therapy.
Language
English
DOI
doi:10.7939/R3TB0R
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
Citation for previous publication
DeSilva A, Wuest M, Wang M, Hummel J, Mossman K, Wuest F, Hitt M. 2012. Comparative functional evaluation of immunocompetent mouse breast cancer models established from PyMT-tumors using small animal PET with [18F]FDG and [18F]FLT. Am J Nucl Med Mol Imaging 2(1):88-98.

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