The goal of this application is to effectively and selectively eliminate B-cell derived lymphoma and leukemia cells in cancer patients using a naturally targeted viral vector system. To achieve this goal, the applicant proposes to develop a novel combinatorial gene therapy approach as applied to Burkett B lymphoma based on i) a minimal """"""""gene-less"""""""" B lymphotropic Epstein-Barr virus (miniEBV) vector, and ii) a genetically enhanced """"""""hyper-suicide"""""""" HSV1 thymidine kinase (super-TK). Specifically, the following studies will be undertaken:
Aim 1) To test the efficacy of B lymphotropic miniEBV vectors to deliver and express a suicide gene into human B-cell lymphomas following oncotropic and oncolytic strategies. This approach is based on the transfer of the viral thymidine kinase (TK) gene into B-cell derived lymphomas rendering them sensitive to the prodrug ganciclovir (GCV). For this endeavor miniEBV/sTK will be used to infect B-lymphoma cells in vitro and the transiently infected cells implanted in an animal model to analyze prodrug mediated eradication of the lymphoma using ex vivo protocol.
Aim 2) Development of an in vitro cultured packaging cell system to produce helper-free infectious miniEBV. This will involve cloning the genome of EBV in a BAC based vector to delete the packaging sequence by homologous recombination. In addition, EBV negative cell lines will be evaluated for their permissivity to miniEBV replication and packaging into infectious virions.
Aim 3) Use the results obtained in Aims 1-2 to test the miniEBV system for its efficiency and safety in a SCID-Human lymphoma/leukemia animal model using an in vivo protocol. Pre-established human B-lymphoma in vivo by intravenous (i.v.) injection with this system will be also evaluated in order to eliminate the disseminated B-lymphoma from various organs of the animal.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL064851-02
Application #
6390726
Study Section
Experimental Therapeutics Subcommittee 1 (ET)
Program Officer
Thomas, John
Project Start
2000-09-10
Project End
2004-08-31
Budget Start
2001-09-15
Budget End
2002-08-31
Support Year
2
Fiscal Year
2001
Total Cost
$180,850
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
078861598
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
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