Oncolytic adenovirus compositions
An adenovirus comprising an E1A polypeptide comprising one or more modifications and comprising an E4orf6/7 polypeptide comprising one or more modifications is described. Compositions and kits comprising the modified adenoviruses are also described. Further described is a method of treating a proliferative disorder in a subject comprising administering to the subject an adenovirus comprising the E1A polypeptide comprising one or more modifications and comprising the E4orf6/7 polypeptide comprising one or more modifications.
1. A recombinant adenovirus comprising:
a modified genome comprising:
(i) an adenovirus E1A promoter operably linked to a nucleic acid sequence encoding an E1A protein comprising a modification in a first Rb-binding site; and
(ii) an adenovirus E4 promoter operably linked to an E4orf6/7 gene product coding sequence having a deletion or a modification of only one of the two E4orf6/7 exons, wherein the deletion of the modification impairs E4orf6/7 gene product activity and/or expression; wherein the recombinant adenovirus selectively replicates in Rb-deficient cells.
2. The recombinant adenovirus of claim 1 , wherein the
E1A protein further comprises a modification in a second Rb-binding site.
3. The recombinant adenovirus of claim 1 , wherein the E1A protein modification is selected from:
a deletion of the LXCXE motif;
a deletion of amino acid residues 122-126;
a deletion of amino acid residues 2-11;
a substitution at residue Y47;
a substitution at residue C124; and any combination of the above.
4. The recombinant adenovirus of claim 3 , wherein the modification in the E1A protein is at least one of the substitution at residue Y47, wherein the substitution is a Y47H substitution, and the substitution at residue C124, wherein the substitution is a C124G substitution.
5. The recombinant adenovirus of claim 1 , the modified genome further comprising: a nucleic acid sequence encoding an E4orf1 protein comprising one or more modifications.
6. The recombinant adenovirus of claim 5 , wherein the one or more modifications of the E4orf1 protein are selected from:
a deletion in the C-terminal region;
a deletion of the last four amino acids in the C-terminal region; and
a deletion of residues 125-128.
7. A pharmaceutical composition comprising the recombinant adenovirus of claim 1 and a pharmaceutically acceptable carrier.
8. The recombinant adenovirus of claim 1 , wherein the deletion or the modification of the only one of the two E4orf6/7 exons is a deletion or a modification of the second E4orf6/7 exon.
9. The recombinant adenovirus of claim 1 , the modified genome further comprising a nucleic acid sequence encoding an E4orf6 protein.
10. The recombinant adenovirus of claim 1 , wherein the modification of the E1A protein comprises a deletion of the LXCXE motif within the first Rb-binding site of said E1A protein.
11. The recombinant adenovirus of claim 10 , wherein the deletion or the modification of the only one of the two E4orf6/7 exons is a deletion of the second E4orf6/7 exon.
12. The recombinant adenovirus of claim 11 , the modified genome further comprising a nucleic acid sequence encoding an E4orf6 protein.
13. The recombinant adenovirus of claim 12 , wherein the adenovirus is serotype 5.
14. A pharmaceutical composition comprising the recombinant adenovirus of claim 13 and a pharmaceutically acceptable carrier.
15. The recombinant adenovirus of claim 10 , wherein the deletion or the modification of only the one of the two E4orf6/7 exons is a modification of the second E4orf6/7 exon.
16. The recombinant adenovirus of claim 15 , the modified genome further comprising a nucleic acid sequence encoding an E4orf6 protein.
17. The recombinant adenovirus of claim 16 , wherein the adenovirus is serotype 5.
18. A pharmaceutical composition comprising the recombinant adenovirus of claim 17 and a pharmaceutically acceptable carrier.
19. A recombinant adenovirus comprising:
(i) an adenovirus E1A promoter operably linked to a nucleic acid sequence encoding an E1A protein comprising a modification in a first Rb-binding site; and
(ii) an adenovirus E4 promoter operably linked to an E4orf6/7 gene product coding sequence having a deletion or a modification of only one of the two E4orf6/7 exons, wherein the deletion or the modification impairs E4orf6/7 gene product activity and/or expression; and
(ii) a nucleic acid sequence encoding an E4orf1 protein comprising one or more modifications; or a deletion of all of the E4orf1 coding sequence;
wherein the recombinant adenovirus selectively replicates in Rb-deficient cells.
20. The recombinant adenovirus of claim 19 , wherein the E1A protein further comprises a modification in a second Rb-binding site.
21. The recombinant adenovirus of claim 19 , wherein the E1A protein modification is selected from:
a deletion of the LXCXE motif; a deletion of amino acid residues 122-126;
a deletion of amino acid residues 2-11;
a substitution at residue Y47;
a substitution at residue C124; and any combination of the above.
22. The recombinant adenovirus of claim 21 , wherein the modification in the E1A protein is at least one of the substitution at residue Y47, wherein the substitution is a Y47H substitution, and the substitution at residue C124, wherein the substitution is a C124G substitution.
23. The recombinant adenovirus of claim 19 , wherein the deletion or the modification of the only one of the two E4orf6/7 exons is a deletion or a modification of the second E4orf6/7 exon.
24. The recombinant adenovirus of claim 19 , wherein the one or more modifications of the E4orf1 protein are selected from:
a deletion in the C-terminal region;
a deletion of the last four amino acids in the C-terminal region; and
a deletion of residues 125-128.
25. A pharmaceutical composition comprising the recombinant adenovirus of claim 19 and a pharmaceutically acceptable carrier.
26. The recombinant adenovirus of claim 19 , the modified genome further comprising a nucleic acid sequence encoding an E4orf6 protein.