Gliobastoma is the most devastating brain tumor in adults and none of the conventional anticancer therapies has been successful in significantly prolonging the lifespan of the patients. More than 80% of gliomas in situ express high amounts of a mutated IL-13 receptor (IL13a2R), different from the one expressed in normal cells. A chimeric toxin consisting of IL-13 fused to a mutated Pseudomonas exotoxin, (PE) has been shown to kill IL-13a2R expressing cells. PE lacks its endogenous protein binding domain, which allows it to bind to the ubiquitous expressed ct2-macroglobulin receptor, but retains its ability to inhibit protein synthesis, which leads to cell death. Thus, its fusion to IL-13 retargets the cytotoxic molecule to IL13a2R-expressing glioma cells. Mutants of IL-13, such as IL13.E13K (mulL-13), have shown negligible binding to normal cells and higher affinity to the glioma-specific IL13a2R when compared to IL-13. However, the systemic treatment with the chimeric toxin requires high concentrations that can induce adverse side effects while the intratumoral treatment requires repeated administration due to the short half life of the toxin. In spite of these limitations this approach is currently considered one of the most exciting novel treatments for this devastating disease, and has provided very encouraging results in early clinical trials performed at various clinical centers. We hypothesize that intratumoral administration of a high-capacity adenoviral vector (HC- Ad) expressing the chimeric protein mulL-13 fused to PE, to human glioma bearing nude mice, will lead to effective intratumoral concentration of the chimeric toxin, killing human glioma cells but preserving normal brain tissue. HC-Ad vectors have the advantage of high cloning capacity and, due to their minimal antigenicity, they elicit stable transgene expression in vivo with low toxicity. To increase the specificity of our approach, we will introduce into the vector the cDNA of mutated IL-4, (mulL4). mulL4 acts as an antagonist, binding to IL13R/IL4R present in normal cells, but it does not interact with IL13a2R expressed on glioma cells, nor does it affect the cytotoxic effects of the chimeric toxin in GBM cells. To further improve the safety of this treatment we incorporated the regulatory TetON system into the vector that allows tight regulation of expression of the therapeutic transgene, by switching it """"""""on"""""""" and """"""""off"""""""" as and when needed by the addition or withdrawal of doxycycline (DOX). Our previous data provide strong evidence that HC-Ad vectors provide long term, regulated expression of the chimeric toxin even in the presence of systemic anti-adenoviral immunity as present in humans. Thus, we hypothesize that the targeted glioma gene therapy approach proposed will elicit high therapeutic efficacy in the absence of adverse side effects. ? ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
1F32NS058156-01A1
Application #
7331147
Study Section
Special Emphasis Panel (ZRG1-F01-N (20))
Program Officer
Fountain, Jane W
Project Start
2007-07-01
Project End
2010-06-30
Budget Start
2007-07-01
Budget End
2008-06-30
Support Year
1
Fiscal Year
2007
Total Cost
$49,646
Indirect Cost
Name
Cedars-Sinai Medical Center
Department
Type
DUNS #
075307785
City
Los Angeles
State
CA
Country
United States
Zip Code
90048
VanderVeen, Nathan; Paran, Christopher; Appelhans, Ashley et al. (2014) Marmosets as a preclinical model for testing ""off-label"" use of doxycycline to turn on Flt3L expression from high-capacity adenovirus vectors. Mol Ther Methods Clin Dev 1:
Candolfi, Marianela; Yagiz, Kader; Wibowo, Mia et al. (2014) Temozolomide does not impair gene therapy-mediated antitumor immunity in syngeneic brain tumor models. Clin Cancer Res 20:1555-1565
Mineharu, Yohei; Muhammad, A K M Ghulam; Yagiz, Kader et al. (2012) Gene therapy-mediated reprogramming tumor infiltrating T cells using IL-2 and inhibiting NF-?B signaling improves the efficacy of immunotherapy in a brain cancer model. Neurotherapeutics 9:827-43
Candolfi, Marianela; King, Gwendalyn D; Yagiz, Kader et al. (2012) Plasmacytoid dendritic cells in the tumor microenvironment: immune targets for glioma therapeutics. Neoplasia 14:757-70
Castro, Maria G; Candolfi, Marianela; Kroeger, Kurt et al. (2011) Gene therapy and targeted toxins for glioma. Curr Gene Ther 11:155-80
Candolfi, Marianela; Curtin, James F; Yagiz, Kader et al. (2011) B cells are critical to T-cell-mediated antitumor immunity induced by a combined immune-stimulatory/conditionally cytotoxic therapy for glioblastoma. Neoplasia 13:947-60
Candolfi, Marianela; Kroeger, Kurt M; Xiong, Weidong et al. (2011) Targeted toxins for glioblastoma multiforme: pre-clinical studies and clinical implementation. Anticancer Agents Med Chem 11:729-38
Puntel, Mariana; Muhammad, A K M G; Candolfi, Marianela et al. (2010) A novel bicistronic high-capacity gutless adenovirus vector that drives constitutive expression of herpes simplex virus type 1 thymidine kinase and tet-inducible expression of Flt3L for glioma therapeutics. J Virol 84:6007-17
Candolfi, Marianela; Xiong, Weidong; Yagiz, Kader et al. (2010) Gene therapy-mediated delivery of targeted cytotoxins for glioma therapeutics. Proc Natl Acad Sci U S A 107:20021-6
Larocque, Daniel; Sanderson, Nicholas S R; Bergeron, Josee et al. (2010) Exogenous fms-like tyrosine kinase 3 ligand overrides brain immune privilege and facilitates recognition of a neo-antigen without causing autoimmune neuropathology. Proc Natl Acad Sci U S A 107:14443-8

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