Use of adenoviral E4 reading frames to improve expression of a gene of interest

A method for improving the expression and/or persistence of expression of a gene of interest in a mammal, said method comprising:

• constructing an expression vector comprising (i), said gene of interest operably linked to regulatory elements that control expression of said gene, and (ii) a polynucleotide sequence consisting of a combination of the adenoviral E4 ORF3 with at least one of the adenoviral E4 open reading frame selected from the group consisting of ORF4, ORF3/4, ORF6/7, ORF6 and ORF7, with the proviso that said expression vector does not comprise any portion of an adenovirus E3 region;
• and
• administering a therapeutically effective amount of said expression vector to a mammal in need thereof.

A method for improving the expression and/or persistence of expression of a gene of interest in a mammal, said method comprising:

• constructing an E1 and E3 deleted adenoviral vector comprising said gene of interest, operably linked to regulatory elements that control expression of said gene, wherein said adenoviral vector is further deleted in the E4 region but retains E4 ORF3 and at least one of the E4 adenoviral open reading frame selected from the group consisting of ORF4, ORF3/4, ORF6/7, ORF6 and ORF7, and
• administering a therapeutically effective amount of said adenoviral vector to a mammal in need thereof.

A method for improving the expression and/or persistence of expression of a gene of interest in lung tissues of a mammal, said method comprising:

• constructing an expression vector comprising (i) said gene of interest, operably linked to regulatory elements that control expression of said gene into said tissue, and (ii) a polynucleotide sequence consisting of a combination of the adenoviral E4 ORF3 with the adenoviral E4 open reading frames ORF6 and/or ORF6/7 or ORF4 and/or ORF3/4 or ORF1, ORF2 and ORF4, said recombinant vector does not further comprise any portion of an adenovirus E3 region, and
• administering a therapeutically effective amount of said expression vector to a mammal in need thereof.

A method for improving the expression and/or persistence of expression of a gene of interest in lung tissues of a mammal, said method comprising:

• constructing an E1 and E3 deleted adenoviral vector comprising said gene of interest, operably linked to regulatory elements that control expression of said gene, wherein said adenoviral vector is partially deleted in the E4 region but specifically retains E4 ORF3 and further E4 open reading frames ORF4 and ORF3/4, or ORF6 and ORF6/7, and
• administering a therapeutically effective amount of said adenoviral vector to a mammal in need thereof, wherein said regulatory elements comprise the early cytomegalovirus promoter.

A method for improving the expression and/or persistence of expression of a gene of interest in the liver tissue of a mammal, said method comprises:

• constructing an expression vector comprising (i) said gene of interest, operably linked to regulatory elements that control expression of said gene into said tissue, and (ii) a polynucleotide sequence consisting of a combination of the adenoviral E4 ORF3 with the adenoviral E4 open reading frames ORF4 and/or ORF314 with the proviso that said expression vector does not comprise any portion of the adenovirus E3 region, and
• administering a therapeutically effective amount of said expression vector to a mammal in need thereof.

A method for reducing hepatotoxicity in a mammal subsequent to the administration to said mammal of an E1 and E3-deleted adenoviral vector comprising a gene of interest placed under the control of the early cytomegalovirus promoter, wherein said adenoviral vector is further partially deleted in the E4 region and specifically retains the adenoviral E4 open reading frame:

• (i) ORF3, or
• (ii) ORF3, ORF4 and ORF3/4,
• wherein hepatotoxicity following administration of said adenoviral vector is reduced in comparison to that observed following administration of an adenoviral vector comprising the complete E4 region.

A method for improving the expression and/or persistence of expression of a gene of interest in the liver tissue of a mammal, said method comprises:

• constructing an E1 and E3 deleted adenoviral vector comprising said gene of interest, operably linked to regulatory elements that control expression of said gene, wherein said adenoviral vector is further deleted in the E4 region but specifically retains E4 ORF3 with E4 ORF4 and ORF3/4,
• administering a therapeutically effective amount of said adenoviral vector to a mammal in need thereof, wherein said regulatory elements comprise the early cytomegovirus.