Gene transfer for treating a connective tissue of a mammalian host

Abstract

Methods for treating a connective tissue disorder by introducing at least one gene encoding a product into at least one target cell of a mammalian host for use in treating the mammalian host are disclosed. These methods include employing recombinant techniques to produce a vector molecule containing the DNA sequence encoding for the product and infecting the target cell of the mammalian host using the vector. The injection can be done in vivo, by directly injecting the vector into the host, or can be done in vitro by transfecting a population of cultured target cells with the vector and transplanting them each into the host. Nonviral means can also be used to introduce the DNA sequence to the host. Administration of more than one gene of interest results in an enhanced therapeutic benefit. Also disclosed is a method for treating a connective tissue disorder by introducing at least one gene encoding a product into at least one target cell of a joint of a host for use in treating multiple joints of the host. Injection of a vector molecule containing the DNA sequence encoding for a product of interest, or non-viral introduction of such a DNA sequence, to one joint of a mammalian host results in a therapeutic benefit in that joint as well as other joints in the host.

What is claimed is:  
     
         1 . A method for treating a connective tissue disorder comprising: 
 a) generating a recombinant vector that comprises at least one DNA sequence encoding one or more genes of interest; and    b) infecting a population of target cells in vivo in a host with said recombinant vector, such that subsequent expression of said gene or genes within said host reduces at least one deleterious joint pathology or indicia of inflammation normally associated with a connective tissue disorder; wherein said gene of interest is one or more therapeutic genes selected from the group consisting of: interleukin-1 receptor antagonist protein; a LacZ marker gene; soluble IL-1 receptor; soluble TNF-α receptor; a proteinase inhibitor; a cytokine; CTLA4; FasL; and biologically active derivatives or fragments of these genes.    
     
     
         2 . The method of  claim 1 , wherein said target cell is selected from the group consisting of connective tissue cells and non-connective tissue cells.  
     
     
         3 . The method of  claim 2 , wherein said connective tissue cells are selected from the group consisting of synovium, cartilage, tendon, ligament, skin, meniscus, bone, and intervertebral disc cells, and said non-connective tissue cells are selected from the group consisting of hematopoietic progenitor cells, stromal cells, bone marrow cells, myoblasts, leukocytes, lymphoid cells and myeloid cells.  
     
     
         4 . The method of  claim 3 , wherein said cytokine is one or more members selected from the group consisting of IL-4, IL-10, IL-13, growth factor, and BMP.  
     
     
         5 . The method of  claim 4 , wherein said BMP is selected from the group consisting of BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8 and BMP-9.  
     
     
         6 . The method of  claim 5 , wherein said BMP is selected from the group consisting of BMP-2 and BMP-7.  
     
     
         7 . The method of  claim 3 , wherein said cytokine is vIL-10.  
     
     
         8 . The method of  claim 3 , wherein said cytokine is growth hormone.  
     
     
         9 . The method of  claim 4 , wherein said growth factor is selected from the group consisting of IGF, FGF and TGF.  
     
     
         10 . The method of  claim 9 , wherein said growth factor is selected from the group consisting of IGF-1 and IGF-2.  
     
     
         11 . The method of  claim 3 , wherein said soluble interleukin-1 receptor is selected from the group consisting of soluble interleukin-1 receptor Type I and soluble interleukin-1 receptor Type II.  
     
     
         12 . The method of  claim 3 , wherein said soluble TNF-α receptor is selected from the group consisting of soluble TNF-α receptor Type I and soluble TNF-α receptor Type II.  
     
     
         13 . The method of  claim 3 , wherein said proteinase inhibitor is selected from the group consisting of TIMP-1, TIMP-2, TIMP-3, TIMP-4, PAIs and serpins.  
     
     
         14 . The method of  claim 1 , wherein said recombinant vector is selected from the group consisting of a viral vector and a non-viral vector.  
     
     
         15 . The method of  claim 14 , wherein said viral vector is selected from the group consisting of a retroviral vector, an adeno-associated viral vector, adenovirus and a herpes viral vector.  
     
     
         16 . The method of  claim 15 , including employing a retrovirus selected from the group consisting MFG and pLJ.  
     
     
         17 . The method of  claim 14 , wherein said recombinant vector is a plasmid DNA vector.  
     
     
         18 . The method of  claim 1 , wherein infection is accomplished by intraarticular injection of the recombinant vector.  
     
     
         19 . The method of  claim 18 , wherein the recombinant vector is an adenoviral vector and the gene encodes for a member selected from the group consisting of IRAP; soluble interleukin-1 receptor; soluble TNF-α receptor; and a cytokine selected from the group consisting of IL-10 and vIL-10.  
     
     
         20 . The method of  claim 18 , wherein the recombinant vector is one or more adenoviral vectors containing genes encoding soluble TNF-α receptor and soluble interleukin-1 receptor.  
     
     
         21 . The method of  claim 1 , wherein the target cells are infected in an area selected from the group consisting of a joint space, bone marrow and bloodstream of said host.  
     
     
         22 . A method for treating a connective tissue disorder comprising: 
 introducing at least one DNA sequence encoding one or more genes of interest into at least one target cell of a host by employing non-viral means selected from the group consisting of at least one liposome, calcium phosphate, electroporation, DEAE-dextran, and direct injection of naked DNA, such that subsequent expression of said gene or genes within said host reduces at least one deleterious joint pathology or indicia of inflammation normally associated with a connective tissue disorder; wherein said gene of interest is one or more therapeutic genes selected from the group consisting of: interleukin-1 receptor antagonist protein; a LacZ marker gene; soluble IL-1 receptor; soluble TNF-α receptor; a proteinase inhibitor; a cytokine; CTLA4; FasL; and biologically active derivatives or fragments of these genes.    
     
     
         23 . The method of  claim 22 , including employing a liposome selected from the group consisting of DC-cholesterol and SF-cholesterol.  
     
     
         24 . A method for treating a connective tissue disorder comprising: 
 a) generating a recombinant vector that comprises at least one DNA sequence encoding one or more genes of interest;    b) infecting a population of target cells in vivo in a first joint of a host with said recombinant vector, such that subsequent expression of said gene or genes in said host reduces at least one deleterious joint pathology or indicia of inflammation normally associated with a connective tissue disorder both in said first joint and in one or more additional joints in the host; wherein said gene of interest is one or more therapeutic genes selected from the group consisting of: interleukin-1 receptor antagonist protein; a LacZ marker gene; soluble IL-1 receptor; soluble TNF-α receptor; a proteinase inhibitor; a cytokine; CTLA4; FasL; and biologically active derivatives or fragments of these genes.    
     
     
         25 . The method of  claim 24 , wherein said target cell is selected from the group consisting of connective tissue cells and non-connective tissue cells.  
     
     
         26 . The method of  claim 25 , wherein said connective tissue cells are selected from the group consisting of synovium, cartilage, tendon, ligament, skin, meniscus, bone, and intervertebral disc cells, and said non-connective tissue cells are selected from the group consisting of hematopoietic progenitor cells, stromal cells, bone marrow cells, myoblasts, leukocytes, lymphoid cells and myeloid cells.  
     
     
         27 . The method of  claim 26 , wherein said cytokine is one or more members selected from the group consisting of IL-4, IL-10, IL-13, growth factor, and BMP.  
     
     
         28 . The method of  claim 27 , wherein said BMP is selected from the group consisting of BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8 and BMP-9.  
     
     
         29 . The method of  claim 28 , wherein said BMP is selected from the group consisting of BMP-2 and BMP-7.  
     
     
         30 . The method of  claim 26 , wherein cytokine is vIL-10.  
     
     
         31 . The method of  claim 26 , wherein said cytokine is growth hormone.  
     
     
         32 . The method of  claim 27 , wherein said growth factor is selected from the group consisting of IGF, FGF and TGF.  
     
     
         33 . The method of  claim 32 , wherein said growth factor is selected from the group consisting of IGF-1 and IGF-2.  
     
     
         34 . The method of  claim 26 , wherein said soluble interleukin-1 receptor is selected from the group consisting of soluble interleukin-1 receptor Type I and soluble interleukin-1 receptor Type II.  
     
     
         35 . The method of  claim 26 , wherein said soluble TNF-α receptor is selected from the group consisting of soluble TNF-α receptor Type I and soluble TNF-α receptor Type II.  
     
     
         36 . The method of  claim 26 , wherein said proteinase inhibitor is selected from the group consisting of TIMP-1, TIMP-2, TIMP-3, TIMP-4, PAIs and serpins.  
     
     
         37 . The method of  claim 24 , wherein said recombinant vector is selected from the group consisting of a viral vector and a non-viral vector.  
     
     
         38 . The method of  claim 37 , wherein said viral vector is selected from the group consisting of a retroviral vector, an adeno-associated viral vector, adenovirus and a herpes viral vector.  
     
     
         39 . The method of  claim 38 , including employing a retrovirus selected from the group consisting MFG and pLJ.  
     
     
         40 . The method of  claim 37 , wherein said recombinant vector is a plasmid DNA vector.  
     
     
         41 . The method of  claim 24 , wherein infection is accomplished by intraarticular injection of the recombinant vector.  
     
     
         42 . The method of  claim 41 , wherein the recombinant vector is an adenoviral vector and the gene encodes for a member selected from the group consisting of IRAP; soluble interleukin-1 receptor; soluble TNF-α receptor; and a cytokine selected from the group consisting of IL-10 and vIL-10.  
     
     
         43 . The method of  claim 41 , wherein the recombinant vector is one or more adenoviral vectors containing genes encoding soluble TNF-α receptor and soluble interleukin-1 receptor.  
     
     
         44 . The method of  claim 24 , wherein the target cells are infected in an area selected from the group consisting of a joint space, bone marrow and bloodstream of said host.  
     
     
         45 . A method for treating a connective tissue disorder comprising introducing at least one DNA sequence encoding one or more genes of interest into at least one target cell in vivo in a first joint of a host by employing non-viral means selected from the group consisting of at least one liposome, calcium phosphate, electroporation, DEAE-dextran, and direct injection of naked DNA, such that subsequent expression of said gene or genes in said host reduces at least one deleterious joint pathology or indicia of inflammation normally associated with a connective tissue disorder both in the first joint and in one or more additional joints in the host; wherein said gene of interest is one or more therapeutic genes selected from the group consisting of: interleukin-1 receptor antagonist protein; a LacZ marker gene; soluble IL-1 receptor; soluble TNF-α receptor; a proteinase inhibitor; a cytokine; CTLA4; FasL; and biologically active derivatives or fragments of these genes.  
     
     
         46 . The method of  claim 22 , including employing a liposome selected from the group consisting of DC-cholesterol and SF-cholesterol.  
     
     
         47 . The method of  claim 15 , including employing a high titer concentration of retroviral vector.  
     
     
         48 . The method of  claim 38 , including employing a high titer concentration of retroviral vector.