Although bacillus Calmette-Guerin (BCG) is the most effective adjuvant treatment for human superficial transitional cell carcinoma (TCC) of the bladder, it is ineffective in up to one third of the cases and it is associated with serious toxicity. The goals of this proposal are to improve the efficacy and reduce the toxicity of BCG by genetically engineering BCG to express and secrete powerful immunoregulatory cytokine proteins. This proposal has three specific aims:
Specific Aim #1 : To engineer recombinant BCG (rBCG) to express and secrete immunoregulatory proteins. Novel genes will be introduced into BCG using a plasmid-based transfection system developed. Specific cytokine genes have been selected based on their potential to complement ongoing cytokine responses (TNF-alpha), antagonize specific cytokine responses (IL-4, IL- 10), or recruit accessory antitumor mechanisms (GM-CSF, IFN-alpha, IL-4). In addition, the Lac Z (beta-galactosidase) gene will be used as a reporter gene to track and quantify rBCG during in vitro and in vivo use. The Herpes Simplex Virus I-thymidine kinase (HSV-tk) gene will be evaluated as a potential """"""""suicide"""""""" gene to eradicate rBCG using the anti- herpetic drug acyclovir.
Specific Aim #2 : To improve our understanding of cytokine pathways elicited by rBCG that have the capacity to modulate antitumor responses. It is important to investigate the cytokine pathways elicited by rBCG so that rational strategies can be formulated to maximize cellular and cytokine responses beneficial to the elimination of TCC while minimizing unproductive or deleterious responses. Endogenous cytokine production from host splenocytes stimulated by rBCG will be measured using sensitive ELISA assays and semi-quantitative Northern blotting. Particular cytokine-BCG combinations will be identified that antagonize or enhance specific splenocyte responses. The specialized immune cell types responsible for these responses will be determined by depleting splenocytes of specific subsets or by replacing them with splenocytes derived from animals with well defined functional immune defects.
Specific Aim #3 : To assess the ability of cytokine rBCG to enhance the immunotherapy of experimental bladder cancer in animal models. Cytokine rBCGs will be tested for activity against TCC in both mouse and rat tumor models. The rat TCC model will provide an independent assessment of rBCG efficacy against the most clinically significant form of bladder cancer, carcinoma-in-situ (CIS).

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29CA064230-05
Application #
2733092
Study Section
Experimental Therapeutics Subcommittee 1 (ET)
Program Officer
Yovandich, Jason L
Project Start
1994-07-01
Project End
1999-12-31
Budget Start
1998-07-01
Budget End
1999-12-31
Support Year
5
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Beth Israel Deaconess Medical Center
Department
Type
DUNS #
076593722
City
Boston
State
MA
Country
United States
Zip Code
02215
O'Donnell, Michael A; Luo, Yi; Hunter, Sharon E et al. (2004) Interleukin-12 immunotherapy of murine transitional cell carcinoma of the bladder: dose dependent tumor eradication and generation of protective immunity. J Urol 171:1330-5
O'Donnell, Michael A; Luo, Yi; Hunter, Sharon E et al. (2004) The essential role of interferon-gamma during interleukin-12 therapy for murine transitional cell carcinoma of the bladder. J Urol 171:1336-42
Nadler, R; Luo, Y; Zhao, W et al. (2003) Interleukin 10 induced augmentation of delayed-type hypersensitivity (DTH) enhances Mycobacterium bovis bacillus Calmette-Guerin (BCG) mediated antitumour activity. Clin Exp Immunol 131:206-16
Luo, Yi; Chen, Xiaohong; O'Donnell, Michael A (2003) Role of Th1 and Th2 cytokines in BCG-induced IFN-gamma production: cytokine promotion and simulation of BCG effect. Cytokine 21:17-26
Luo, Y; Chen, X; Han, R et al. (2001) Recombinant bacille Calmette-Guerin (BCG) expressing human interferon-alpha 2B demonstrates enhanced immunogenicity. Clin Exp Immunol 123:264-70
Pavlovich, C P; Kraling, B M; Stewart, R J et al. (2000) BCG-induced urinary cytokines inhibit microvascular endothelial cell proliferation. J Urol 163:2014-21
Luo, Y; Chen, X; Szilvasi, A et al. (2000) Co-expression of interleukin-2 and green fluorescent protein reporter in mycobacteria: in vivo application for monitoring antimycobacterial immunity. Mol Immunol 37:527-36
O'Donnell, M A; Luo, Y; Chen, X et al. (1999) Role of IL-12 in the induction and potentiation of IFN-gamma in response to bacillus Calmette-Guerin. J Immunol 163:4246-52
Luo, Y; Chen, X; Downs, T M et al. (1999) IFN-alpha 2B enhances Th1 cytokine responses in bladder cancer patients receiving Mycobacterium bovis bacillus Calmette-Guerin immunotherapy. J Immunol 162:2399-405
Alexandroff, A B; Jackson, A M; O'Donnell, M A et al. (1999) BCG immunotherapy of bladder cancer: 20 years on. Lancet 353:1689-94

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