Mixed-cell gene therapy

Abstract

The subject invention is directed to a mixed cell composition to generate a therapeutic protein at a target site by providing a first population of mammalian cells transfected or transduced with a gene that is sought to be expressed, and a second population of mammalian cells that have not been transfected or transduced with the gene, wherein endogenously existing forms of the second population of mammalian cells are decreased at the target site, and wherein generation of the therapeutic protein by the first population of mammalian cells at the target site stimulates the second population cells to induce a therapeutic effect.

1 - 18 . (canceled) 
     
     
         19 . A method of generating a therapeutic protein at a target site in a mammal comprising:
 a) generating a recombinant vector comprising a DNA sequence encoding the therapeutic protein operatively linked to a promoter;   b) transfecting or transducing a population of cells in vitro with said recombinant vector; and   c) injecting a mixed cell composition comprising protein generating effective amount of (i) a first population of cells transfected or transduced with the gene; (ii) a second population of cells that have not been transfected or transduced with the gene; and (iii) a pharmaceutically acceptable carrier thereof, into the target site, wherein endogenously existing forms of the second population of mammalian cells are decreased at the target site, and wherein generation of the therapeutic protein by the first population of mammalian cells at the target site stimulates the second population cells to induce a therapeutic effect.   
     
     
         20 . The method according to  claim 19 , comprising a method of generating hyaline cartilage in a mammal comprising:
 a) generating a recombinant vector comprising a DNA sequence encoding transforming growth factor β (TGF-β) or bone morphogenic protein (BMP) operatively linked to a promoter;   b) transfecting or transducing a population of fibroblast or chondrocyte cells in vitro with said recombinant vector; and   c) injecting an injectable mixed cell composition comprising hyaline cartilage-generating effective amount of (i) a first population of fibroblast or chondrocyte cells transfected or transduced with a gene encoding TGF-β or BMP; (ii) a second population of fibroblast or chondrocyte cells that have not been transfected or transduced with a gene encoding TGF-β or BMP; and (iii) a pharmaceutically acceptable carrier thereof, into a joint space of a mammal such that expression of the DNA sequence encoding TGF-β or BMP within the joint space occurs resulting in the generation of hyaline cartilage in the joint space.   
     
     
         21 . The method according to  claim 20 , wherein said gene is TGF-β1, TGF-β2, TGF-β3, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, or BMP-7. 
     
     
         22 . The method according to  claim 21 , wherein said gene is TGF-β1 or BMP-2. 
     
     
         23 . The method according to  claim 20 , wherein said ratio of the second population of fibroblast or chondrocyte cells that have not been transfected or transduced with a gene encoding TGF-β or BMP to the first population of fibroblast or chondrocyte cells that have been transfected or transduced with a gene encoding TGF-β or BMP is from about 1-20 to 1. 
     
     
         24 . The method according to  claim 23 , wherein said ratio is from about 3-10 to 1. 
     
     
         25 . The method according to  claim 24 , wherein said ratio is from about 10 to 1. 
     
     
         26 . The method according to  claim 20 , wherein the first population of fibroblast or chondrocyte cells transfected or transduced with a gene encoding TGF-β or BMP is irradiated. 
     
     
         27 . The method according to  claim 20 , wherein the first population of fibroblast or chondrocyte cells transfected or transduced with the gene encoding TGF-β or BMP and the second population of fibroblast or chondrocyte cells not transfected or transduced with a gene encoding TGF-β or BMP are syngeneic with respect to the host recipient. 
     
     
         28 . The method according to  claim 20 , wherein the first population of fibroblast or chondrocyte cells transfected or transduced with the gene encoding TGF-β or BMP and the second population of fibroblast or chondrocyte cells not transfected or transduced with a gene encoding TGF-β or BMP are allogeneic with respect to the host recipient. 
     
     
         29 . The method according to  claim 20 , wherein the first population of fibroblast or chondrocyte cells transfected or transduced with the gene encoding TGF-β or BMP and the second population of fibroblast or chondrocyte cells not transfected or transduced with a gene encoding TGF-β or BMP are xenogeneic with respect to the host recipient. 
     
     
         30 . The method of  claim 20 , wherein said recombinant vector is a viral vector. 
     
     
         31 . The method of  claim 20 , wherein said recombinant vector is a plasmid vector. 
     
     
         32 . The method of  claim 20 , wherein said cells are stored prior to transplantation. 
     
     
         33 . The method of  claim 32 , wherein said cells are stored in a cryopreservative prior to transplantation. 
     
     
         34 . The method of  claim 20 , wherein said transfection or transduction is accomplished by liposome encapsulation, calcium phosphate coprecipitation, electroporation, DEAE-dextran mediation or virus mediation. 
     
     
         35 . A method of treating osteoarthritis comprising:
 a) generating a recombinant vector comprising a DNA sequence encoding transforming growth factor β (TGF-β) or bone morphogenic protein (BMP) operatively linked to a promoter;   b) transfecting or transducing a population of fibroblast or chondrocyte cells in vitro with said recombinant vector; and   c) injecting an injectable mixed cell composition comprising hyaline cartilage-generating and osteoarthritis treating effective amount of,
 (i) a first population of fibroblast or chondrocyte cells transfected or transduced with a gene encoding TGF-β or BMP; 
 (ii) a second population of fibroblast or chondrocyte cells that have not been transfected or transduced with a gene encoding TGF-β or BMP; and 
 (iii) a pharmaceutically acceptable carrier thereof that is not a non-living three dimensional structure into a joint space of a mammal such that expression of the DNA sequence encoding TGF-β or BMP within the joint space occurs resulting in the generation of bone and cartilage tissue in the joint space. 
   
     
     
         36 - 38 . (canceled)