FOMDWebPartPage

Experimental Oncology

Mary Hitt

Chair, Cancer Sciences Graduate Coordinating Committee & Associate Professor
Experimental Oncology
Department of Oncology
University of Alberta
5-142H Katz Group Centre
114 Street & 87 Avenue
Edmonton, Alberta T6G 2E1

Tel: 780.492.6351
mary.hitt@ualberta.ca

Profile

Tumor-targeted cancer gene therapy, viral oncolysis, and mechanisms of viral transformation.
 
Although much progress has been made toward the prevention and therapy of cancer, more Canadians die each year from cancer or cancer-related causes than from any other disease. This has prompted investigators to examine new anti-cancer agents and new approaches to optimize administration of these agents. Many of these agents can have significant side effects when active in normal tissues.
 
One area of research in my laboratory focuses on the development of gene therapy vectors that minimize these side effects without compromising anticancer activity. Currently we are investigating adenovirus vectors carrying modified capsid proteins and/or tissue-specific regulatory elements to target expression of cytotoxic genes specifically to the tumor. As an alternative to transfer of therapeutic genes to tumor cells, we are investigating the ability of replicating viruses (both adenovirus and poxviruses) to specifically eliminate tumor cells in vitro and in tumor models. A third line of study involves the investigation of mechanisms of viral transformation as a model for early stages of tumorigenesis.
 
Research Interests
 
Vector targeting. My laboratory primarily uses the adenovirus (Ad) vector system to transduce cancer cells with potentially therapeutic genes. The advantages of Ad are its high efficiency of transduction, ease of propagation and purification, stability, and the wide range of cell types, both proliferating and quiescent, that can be infected by the virus. Unfortunately, many tumor cells display reduced levels of the receptors required for Ad infection, resulting in greater gene transfer to normal cells than tumor cells under normal conditions. This problem can be circumvented by modifying the virus capsid so that it recognizes alternate receptors on the tumor cell surface. In collaboration with Dr. Frank Graham at McMaster University, we have recently generated a vector that binds to ErbB3 and ErbB4 receptors that are frequently over-expressed on tumor cells. This vector modification enhances in vitro transduction of cancer cells but not normal cells (MacLeod et al., 2012). Whether this modification results in reduced toxicity to normal tissues in vivo is currently under investigation in my lab.
 
Transcriptional targeting. Another approach for reducing toxicity to normal cells is to target expression of the therapeutic gene using tissue-specific or tumor-specific promoters. In particular, we have focused on regulatory sequences derived from the mammaglobin gene which is expressed at high levels in mammary carcinoma cells, to a lesser extent in normal mammary cells, and is undetectably expressed in nearly all other normal cells (Shi et al. 2005, 2006). We have recently isolated a 340 bp minimal promoter sequence immediately 5' to the mammaglobin open reading frame that appears to be responsible for the high degree of specificity of expression. In addition, we have identified an upstream enhancer element that boosts expression levels by at least 10-fold in breast cancer cells.
 
Therapeutic genes. The utility of Ad vectors for the transfer of anti-cancer genes directly to tumors has been investigated by numerous labs over the past 20 years. Some of these vectors, including those encoding the cytokines IL-2, IL-12 and the costimulatory factor B7-1 (CD80) developed by myself and colleagues at McMaster University and University of Toronto, have been used in clinical trials for cancer therapy. Results of both pre-clinical and clinical trials demonstrate the potential of Ad vectors, but also indicate that the genes being transferred are not potent enough in tumor cell killing. Extremely potent death-inducing vectors can be difficult to rescue and amplify due premature killing of 'packaging' cell lines. We are currently developing systems that will attenuate expression of toxic genes in the cell lines used to rescue and propagate vectors, as well as in normal cells. One way to modulate expression exploits the RNA interference pathway. We are investigating whether insertion of sequences targeted by naturally-occurring or introduced microRNAs can facilitate silencing of virus-encoded targeted genes.  We hope to use these new systems to produce vectors encoding pro-apoptotic proteins for tumor-targeted cancer gene therapy. 
 
Viral oncolysis. Viral oncolysis is the selective killing of tumor cells by replicating viruses. One mechanism for such selective killing is to exploit cellular processes that are altered in both infected cells and in tumor cells. Such processes include evasion of innate host immune responses (such as the interferon response), stimulation of DNA synthesis and cell cycling, activation of signaling pathways, and inactivation of tumor suppressors such as p53 and pRb. Viruses can be designed or selected that are unable to replicate in normal cells due to host control of these pathways, yet can replicate efficiently in tumor cells which have lost regulation of these pathways. This category of oncolytic viruses includes adenoviruses from which the E1B and/or VA-RNA genes have been deleted. One E1B protein, E1B-55K binds p53 and sequesters it into an aggresome to prevent its activity. Another E1B product is involved in blocking apoptosis. VA RNAs are known to play an important role in counteracting the interferon response in normal cells, a function which is redundant in many tumor cells. Characterization of VA-RNA deleted viruses is underway (Sharon et al., 2013).
 
In addition, in collaboration with Dr. David Evans (Medical Microbiology and Immunology) we are investigating poxviruses (myxoma virus, Shope fibroma virus and vaccinia) for their oncolytic potential. Leporipoxviruses exhibit a very narrow host range limited to rabbits and hares.  However the host range can be expanded to encompass transformed human cells as a result of the constitutive activation of the protein kinase B/Akt signaling pathways in some tumor cells.  This forms the basis for using myxoma virus as an oncolytic agent.  We are currently testing the efficacy of these viruses in vitro and in mouse tumor models (Irwin et al., in press). Recent experiments suggest that vaccinia virus can be modified to improve its safety as an anticancer agent. This modified vaccinia is under investigation in breast cancer and bladder cancer (Potts et al., 2012) tumor models to test its activity and specificity for tumor cells.
 
Viral transformation. Many of the cellular processes that are subverted at early stages of virus infection are also subverted at early stages of tumorigenesis. In collaboration with Dr. Andy Shaw (Experimental Oncology), we are investigating a novel cellular protein that may play an important role in intracellular trafficking of regulatory molecules during adenoviral replication and transformation.

 

Publications

Chaurasiya, S., Hew, P., Crosley, P., Sharon, D., Potts, K., Agopsowicz, K., Long, M., Shi, C.-X., and Hitt, M.M. (2016). Breast cancer gene therapy using an adenovirus encoding human IL-2 under control of mammoglobin promoter/enhancer sequences. Cancer Gene Therapy 23, 178-187.
 
Chaurasiya, S., and Hitt, M.M. (2016). Adenoviral vector construction 1: Mammalian systems. In “Adenoviral Vectors for Gene Therapy” (D. Curiel, ed.), Second edition, Elsevier, San Diego, CA, USA. pp. 85-112.
 
Forbrich, A., Paproski, R., Hitt, M., and Zemp, R. (2014). Comparing efficiency of micro-RNA and mRNA biomarker liberation with microbubble-enhanced ultrasound exposure. Ultrasound Med Biol. 40(9): 2207-16.
 
Benesch, M.G.K., Tang, X., Maeda, T., Ohhata, A., Zhao, Y.Y., Kok, B.P.C., Dewald, J., Hitt, M., Curtis, J.M., McMullen, T.P.W., and Brindley, D.N. (2014). Inhibition of autotaxin delays breast tumor growth and lung metastasis in mice. FASEB J. 28(6): 2655-66.
 
Irwin, C.R., Favis, N.A., Agopsowicz, K.C., Hitt, M.M., and Evans, D.H. (2013). Myxoma virus oncolytic efficiency can be enhanced through chemical and genetic disruption of the actin cytoskeleton. PLoS ONE 2013 8(12):e84134.
 
Sharon, D., Schümann, M., MacLeod, S.H., McPherson, R., Chaurasiya, S., Shaw, A., and Hitt, M.M. (2013). 2-Aminopurine enhances the oncolytic activity of an E1b-deleted adenovirus in hepatocellular carcinoma cells. PLoS ONE 2013 8(6):e65222.
 
MacLeod, S.H., Elgadi, M. M., ,Bossi, G., Sankar, U., Pisio, A., Agopsowicz, K., Sharon, D., Graham, F.L., and Hitt, M.M. (2012). HER3-targeting of adenovirus by fiber modification increases infection of breast cancer cells in vitro, but not following intratumoral injection in mice. Cancer Gene Therapy 19: 888-98.
 
Gelebart, P., Hegazy, S., Wang, P., Bone, K., Anand, M., Sharon, D., Hitt, M., Ma, Y., and Lai, R. (2012). Abberant expression and biological significance of Sox2, an embryonic stem cell transcriptional factor, in ALK-positive anaplastic large cell lymphoma. Blood Cancer J. Aug 10; 2:e82..
 
Potts, K.G., Hitt, M.M., and Moore, R.B. (2012). Oncolytic viruses in the treatment of bladder cancer. Advances in Urology Volume 2012, Article ID 404581,doi:10.1155/2012/404581.
 
Wang, P., Zhang, J.-D., Wu, F., Ye, X., Sharon, D., Hitt, M., McMullen, T.P, Hegazy, S.A., Gelebart, P., Yang, J., Ma, Y., and Lai, R. (2012). The expression and oncogenic effects of the embryonic stem cell marker SALL4 in ALK-positive anaplastic large cell lymphoma. Cell Signal.24: 1955-1963. 
 
DeSilva, A., Wuest, M., Wang, M., Hummel, J., Mossman, K., Wuest, F., and Hitt, M.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: 88-98.
 
Paproski, R.J., Forbrich, A.E., Wachowicz, K., Hitt, M., and Zemp, R. J. (2011). Tyrosinase as a dual reporter gene for both photoacoustic and magnetic resonance imaging. Biomed Optics Express. 2:771-80.
 
Graham, F.L. and Hitt, M.M. (2007). Adenoviral Vectors in Gene Therapy. In: Encyclopedia of Life Sciences. John Wiley & Sons, Ltd: Chichesterhttp://www.els.net/[DOI: 10.1002/9780470015902.a0005737.pub2]
 
Shi, C.-X., Graham, F.L., and Hitt, M.M. (2006).  A convenient plasmid system for construction of helper-dependent adenoviral vectors and its application for mammaglobin promoter studies.  J. Gene Med.8, 442-51.
 
Hitt, M., Ng, P., and Graham, F.L. (2006). Construction and propagation of human adenovirus vectors. In “Cell Biology: A Laboratory Handbook” (J.E. Celis, ed.), Third edition, Academic Press, San Diego, CA.,Vol. 1, pp. 435-443.
 
See, R.H., Petric, M., Zakhartchouk, A.N., Lawrence, D.J., Mok, C.P.Y., Hogan, R.J., Rowe, T., Zitzow, L.A., Karunakaran, K.P., Hitt, M.M., Graham, F.L., Prevec, L., Mahony, J., Tingle, A.J., Scheifele, D.W., Skowronski, D.M., Patrick, D.M., Babiuk, L.A., Gauldie, J., Voss, T.G., Roper, R.L., Brunham, R.C., and Finlay, B.B. (2006). Comparative evaluation of two SARS vaccine candidates in mice challenged with SARS-coronavirus. J. Gen. Virol.87, 641-50.
 
Santosuosso, M., Zhang, X., McCormick, S., Wang, J., Hitt, M., and Xing, Z. (2005). Mechanisms of mucosal and parenteral tuberculosis vaccinations: adenoviral-based mucosal immunization preferentially elicits sustained accumulation of immune protective CD4 and CD8 T cells within the airway lumen. J. Immunol. 174: 7986-94.
 
Xing, Z., Santosuosso, M., McCormick, S., Yang, T.-C., Millar, J., Hitt, M., Wan, Y., Bramson, J., and Vordermeier, H.M. (2005). Recent advances in the development of adenovirus- and poxvirus-vectored tuberculosis vaccines. Current Gene Therapy 5, 485-492.
 
Sadeghi, H. and Hitt, M.M. (2005). Transcriptional targeting of adenovirus vectors. Current Gene Therapy 5: 411-427.
 
Shi, C.X., Long, M.A., Liu, L., Graham, F.L., Gauldie, J., and Hitt, M.M. (2004). The human SCGB2A2 (mammaglobin-1) promoter/enhancer in a helper-dependent adenovirus vector directs high levels of transgene expression in mammary carcinoma cells but not normal non-mammary cells. Molecular Therapy 10:758-767.