Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6461869

Title:Purging leukemia cells from hematopoietic stem cells
Inventors:Schwarzenberger, Paul; New Orleans, LA, USA
Kolls, Jay; New Orleans, LA, USA
Summary:Described herein are methods of purging leukemia cells from a mixture containing leukemia cells and stem cells, by transforming the mixture with an adenovirus vector having a DNA sequence-encoding toxin and promoter, and allowing the toxin to kill leukemia cells. The invention relates to a gene therapy system which selectively kills leukemia cells in bone marrow, while leaving stem cells unaffected. Further disclosed are methods by which all cells in a mixture of stem cells and leukemia cells are transfected with a high efficiency gene transfer vector carrying a eukaryotic expression construct encoding a toxin gene which is expressed only in leukemia cells, not in stem cells. Also provided are the means by which differential expression of the toxin gene in leukemia cells and in stem cells may be achieved by placing the coding sequence under the control of an appropriate promoter, such as the RSV or the SV40 promoters. Claims of the invention include high gene expression that has been demonstrated in a panel of transformed leukemia cell lines, with no gene expression demonstrated in transformed, CD34-selected, primary human stem cells. Therapeutic applications are directed toward the treatment of leukemia and other cancer patients who may be undergoing autologous bone marrow transplants such breast or lymphoma cancers.
Abstract:A gene therapy system is disclosed that selectively kills leukemia cells in bone marrow, while leaving stem cells unaffected. All cells in a mixture of stem cells and leukemia cells are transfected with a high efficiency gene transfer vector. The vector carries a eukaryotic expression construct encoding a toxin gene. This toxin gene is expressed only in leukemia cells, not in stem cells. Differential expression of the toxin gene in leukemia cells and stem cells may be achieved by placing the coding sequence under the control of an appropriate promoter, such as the RSV promoter or the SV40 promoter. High gene expression has been demonstrated in a panel of transformed leukemia cell lines, but no gene expression in transformed, CD34-selected, primary human stem cells. The treatment will be useful not only for leukemia patients, but also for other cancer patients undergoing autologous bone marrow transplants (e.g., breast or lymphoma cancers).
US Patent Website:Click Here for Full Text of Patent
Title Number:US6461869
Application Number:US1999000357752
Date Filed:20/07/1999
Date Published:08/10/2002
Assignee:Board of Supervisors of Louisiana State University and Agricultural and Mechanical College, Baton Rouge, LA, USA


 
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