Adenovirus vectors for gene therapy

Abstract

Gene Therapy vectors, which are especially useful for cystic fibrosis, and methods for using the vectors are disclosed.

1 . An adenovirus-based gene therapy vector comprising the genome of an adenovirus 2 serotype in which the E1a and E1b regions of the genome, which are involved in early stages of viral replication, have been deleted and replaced by genetic material of interest.  
     
     
         2 . The adenovirus-based gene therapy vector of  claim 1 , wherein the genetic material of interest is DNA encoding cystic fibrosis transmembrane conductance regulator  
     
     
         3 . The adenovirus-based gene therapy vector of  claim 1  further comprising PGK promoter operably linked to the genetic material of interest.  
     
     
         4 . The adenovirus-based gene therapy vector of  claim 2  having substantially the same nucleotide sequence as shown in Table II (SEQ ID NO:3).  
     
     
         5 . An adenovirus-based gene therapy vector comprising adenovirus inverted terminal repeat nucleotide sequences and the minimal nucleotide sequences necessary for efficient replication and packaging and genetic material of interest.  
     
     
         6 . The adenovirus-based gene therapy vector of  claim 5  having the adenovirus 2 sequences shown in FIG. 17.  
     
     
         7 . The adenovirus-based gene therapy vector of  claim 5  further comprising PGK promoter operably linked to the genetic material of interest.  
     
     
         8 . The adenovirus-based gene therapy vector of  claim 5  in which the genetic material of interest is selected from the group consisting of DNA encoding: cystic fibrosis transmembrane conductance regulator, Factor VIII, and Factor IX.  
     
     
         9 . An adenovirus-based gene therapy vector comprising an adenovirus genome which has been deleted for all E4 open reading frames, except open reading fine 6, and additionally comprising genetic material of interest.  
     
     
         10 . The adenovirus-based gene therapy vector of  claim 9  further comprising PGK promoter operably linked to the genetic material of interest.  
     
     
         11 . The adenovirus-based gene therapy vector of  claim 9  in which the E1a and E1b regions of the genome, which are involved in early stages of viral replication, have been deleted.  
     
     
         12 . The adenovirus-based gene therapy vector of  claim 9  in which the E3 region has been deleted.  
     
     
         13 . An adenovirus-based gene therapy vector comprising an adenovirus genome which has been deleted for all E4 open reading frames, except open reading frame 3, and additionally comprising genetic material of interest.  
     
     
         14 . The adenovirus-based gene therapy vector of  claim 13  in which the E1a and E1b regions of the genome, which are involved in early stages of viral replication, have been deleted.  
     
     
         15 . The adenovirus-based gene therapy vector of  claim 13  further comprising PGK promoter operably linked to the genetic material of interest.  
     
     
         16 . The adenovirus-based gene therapy vector of  claim 13  in which the E3 region has been deleted.  
     
     
         17 . A method for treating or preventing cystic fibrosis in a patient comprising administering to the pulmonary airways of the patient, a gene therapy vector comprising DNA encoding cystic fibrosis transmembrane conductance regulator.  
     
     
         18 . The method of  claim 17  wherein the gene therapy vector is an adenovirus-based gene therapy vector comprising the genome of an adenovirus 2 serotype in which the E1a and E1b regions of the genome, which are involved in early stages of viral replication, have been deleted and replaced by DNA encoding cystic fibrosis transmembrane conductance regulator.  
     
     
         19 . The method of  claim 17  wherein the gene therapy vector further comprises PGK promoter operably linked to the DNA encoding cystic fibrosis transmembrane conductance regulator.  
     
     
         20 . The method of  claim 17  wherein the gene therapy vector is an adenovirus-based gene therapy vector comprising adenovirus inverted terminal repeats and the minimal sequences necessary for efficient replication and packaging and DNA encoding cystic fibrosis tranmembrane conductance regulator.  
     
     
         21 . The method of  claim 20  wherein the gene therapy vector further comprises PGK promoter operably linked to the DNA encoding cystic fibrosis transmembrane conductance regulator.  
     
     
         22  The method of  claim 17  wherein the gene therapy vector is an adenovirus-based gene therapy vector comprising an adenovirus genome which has been deleted for all E4 open reading frames, except open reading frame 6, and additionally comprising DNA encoding cystic fibrosis transmembrane conductance regulator.  
     
     
         23 . The method of  claim 22  wherein the gene therapy vector further comprises PGK promoter operably linked to the DNA encoding cystic fibrosis transmembrane conductance regulator.  
     
     
         24 . The method of  claim 17  wherein the gene therapy vector is an adenovirus-based gene therapy vector comprising an adenovirus genome which has been deleted for all E4 open reading frames, except open reading frame 6, and has been deleted for the E1a and E1b regions of the genome, which are involved in early stages of viral replication, and additionally comprising DNA encoding cystic fibrosis tranmembrane conductance regulator.  
     
     
         25 . The method of  claim 24  wherein the gene therapy vector further comprises PGK promoter operably linked to the DNA encoding cystic fibrosis transmembrane conductance regulator.