Claims
- 1. A recombinant adenovirus vector, comprising a recombinant adenovirus vector genome, comprising:
(a) a heterologous nucleic acid sequence; (b) a functional E1 coding region; and (b) a mutation in the 100K coding region, such that said adenovirus vector genome has an impairment in 100K activity.
- 2. The recombinant adenovirus vector of claim 1, wherein said adenovirus expresses less than about 25% of the 100K activity of an adenovirus with a wild-type 100K coding region.
- 3. The recombinant adenovirus vector of claim 1, wherein said adenovirus vector genome expresses no detectable 100K activity.
- 4. The recombinant adenovirus vector of claim 1, wherein said mutation in the 100K coding region comprises a deletion in the 100K coding region.
- 5. The recombinant adenovirus vector of claim 4, wherein said deletion in the 100K coding region comprises a deletion from about nucleotide 24,990 to about nucleotide 25,687 of the adenovirus serotype 5 genome or a corresponding region of the genome of adenoviruses of other serotypes.
- 6. The recombinant adenovirus vector of claim 1, wherein said adenovirus may be propagated to produce new adenovirus particles in a cell that expresses a functional 100K protein.
- 7. The recombinant adenovirus vector of claim 1, wherein said adenovirus may be propagated in a transcomplementing cell in the absence of a helper.
- 8. The recombinant adenovirus vector of claim 1, wherein said adenovirus vector genome comprises a deletion in the E3 coding region.
- 9. The recombinant adenovirus vector of claim 1, wherein said adenovirus vector genome is replicated upon introduction into an adenovirus permissive cell.
- 10. The recombinant adenovirus vector of claim 1, wherein said adenovirus particle is a type 2 or type 5 adenovirus particle.
- 11. The recombinant adenovirus vector of claim 1, wherein said heterologous nucleotide sequence is operatively associated with an expression control sequence.
- 12. The recombinant adenovirus vector of claim 1, wherein said expression control sequence comprises a promoter.
- 13. The recombinant adenovirus vector of claim 12, wherein said promoter is selected from the group consisting of liver-specific, skeletal muscle-specific, cardiac muscle-specific, smooth muscle-specific, diaphragm muscle-specific, prostate-specific, and brain-specific promoters.
- 14. The recombinant adenovirus vector of claim 12, wherein said promoter is a cancer cell specific promoter.
- 15. The recombinant adenovirus vector of claim 12, wherein said promoter is an inducible promoter.
- 16. The recombinant adenovirus vector of claim 1, wherein said E1 coding region is operatively associated with a promoter selected from the group consisting of liver-specific, skeletal muscle-specific, cardiac muscle-specific, smooth muscle-specific, diaphragm muscle-specific, prostate-specific, and brain-specific promoters.
- 17. The recombinant adenovirus vector of claim 1, wherein said E1 coding region is operatively associated with a cancer cell specific promoter.
- 18. The recombinant adenovirus vector of claim 1, wherein said E1 coding region is operatively associated with an inducible promoter.
- 19. The recombinant adenovirus vector of claim 1, wherein said heterologous nucleotide sequence encodes a polypeptide.
- 20. The recombinant adenovirus vector of claim 19, wherein said polypeptide is an industrial enzyme.
- 21. The recombinant adenovirus vector of claim 19, wherein said polypeptide is an adeno-associated virus capsid protein.
- 22. The recombinant adenovirus vector of claim 19, wherein said polypeptide is an adeno-associated virus Rep protein.
- 23. The recombinant adenovirus vector of claim 19, wherein said polypeptide is an immunogenic polypeptide that induces an immune response against a pathogen.
- 24. The recombinant adenovirus vector of claim 23, wherein said immunogenic polypeptide is a cancer cell immunogen.
- 25. The recombinant adenovirus vector of claim 19, wherein said polypeptide is a reporter polypeptide.
- 26. The recombinant adenovirus vector of claim 1, wherein said heterologous nucleotide sequence encodes an antisense RNA.
- 27. The recombinant adenovirus vector of claim 19, wherein said polypeptide is a therapeutic polypeptide.
- 28. The recombinant adenovirus vector of claim 27, wherein said polypeptide is associated with a lysosomal storage disease.
- 29. The recombinant adenovirus vector of claim 27, wherein said polypeptide is associated with a glycogen storage disease.
- 30. The recombinant adenovirus vector of claim 29, wherein said polypeptide is lysosomal acid a-glucosidase.
- 31. The recombinant adenovirus vector of claim 25, wherein said polypeptide is an anti-cancer agent.
- 32. The recombinant adenovirus vector of claim 1, wherein said recombinant adenovirus vector genome is encapsidated within an adenovirus capsid.
- 33. A recombinant adenovirus vector, comprising a recombinant adenovirus vector genome, comprising:
(a) a heterologous nucleic acid sequence; (b) a functional E1a coding region encoding sufficient E1a gene products for adenovirus genomic replication; (c) a mutation in the E1b coding region, such that said adenovirus vector genome has an impairment in p55 activity; and (d) a mutation in the 100K coding region, such that said adenovirus vector genome has an impairment in 100K activity.
- 34. The recombinant adenovirus vector of claim 33, wherein said mutation in the E1b coding region comprises a deletion in the E1b coding region.
- 35. The recombinant adenovirus vector of claim 33, wherein said recombinant adenovirus vector genome is encapsidated within an adenovirus capsid.
- 36. A pharmaceutical formulation comprising the recombinant adenovirus particle of claim 32 or claim 35 in a pharmaceutically acceptable carrier.
- 37. A cultured cell comprising the recombinant adenovirus vector of claim 1 or claim 33.
- 38. The cell of claim 37, wherein said cell is a mammalian cell.
- 39. The cell of claim 37, wherein a nucleic acid sequence encoding a functional adenovirus 100K protein is stably expressed by the cell.
- 40. An isolated nucleic acid sequence comprising the recombinant vector genome of claim 1 or claim 33
- 41. The isolated nucleic acid sequence of claim 40, wherein said isolated nucleic acid sequence is a plasmid.
- 42. A method of producing an adenovirus particle, comprising:
introducing a recombinant adenovirus vector according to claim 1 or claim 33 into a cell that expresses a functional 100K protein under conditions sufficient for replication and packaging of adenovirus particles in the cell; and collecting the adenovirus particles.
- 43. The method of claim 42, wherein a nucleotide sequence encoding a functional adenovirus 100K protein is stably expressed by the cell.
- 44. An adenovirus particle produced by the method of claim 42.
- 45. A method of introducing a nucleic acid sequence into a cell, comprising contacting a cell with a recombinant adenovirus vector according to claim 1 or claim 33 under conditions sufficient for entry of the adenovirus particle into the cell.
- 46. The method of claim 45, wherein the cell is selected from the group consisting of a neuron, a brain cell, a retinal cell, an epithelial cell, a cardiac muscle cell, a smooth muscle cell, a skeletal muscle cell, a diaphragm muscle cell, a pancreatic cell, a liver cell, a fibroblast, an endothelial cell, a germ cell, a lung cell, a prostate cell, a stem cell, and a progenitor cell.
- 47. The method of claim 45, wherein the cell is a cancer cell.
- 48. The method of claim 45, wherein the cell is a mammalian cell.
- 49. The method of claim 45, wherein the adenovirus vector genome is replicated in the cell.
- 50. The method of claim 45, wherein the cell does not provide a functional 100K protein.
- 51. The method of claim 50, wherein accumulation of an adenovirus late gene product in the cell is reduced as compared with an adenovirus vector that expresses a functional 100K protein.
- 52. A method of administering a nucleotide sequence to a subject, comprising administering to a subject a cell according to claim 45 in a pharmaceutically acceptable carrier.
- 53. A method of administering a nucleotide sequence to a subject, comprising administering to a subject a recombinant adenovirus vector according to claim 1 or claim 33 in a pharmaceutically acceptable carrier.
- 54. The method of claim 53, wherein the subject is selected from the group consisting of avian subjects and mammalian subjects.
- 55. The method of claim 54, wherein the subject is a mammalian subject.
- 56. The method of claim 55, wherein the subject is a human subject.
- 57. The method of claim 53 or claim 56, wherein the subject has been diagnosed with lysosomal acid α-glucosidase deficiency.
- 58. The method of claim 53, wherein the subject has cancer.
- 59. The method of claim 55, wherein the recombinant adenovirus vector is administered by a route selected from the group consisting of oral, rectal, transmucosal, transdermal, inhalation, intravenous, subcutaneous, intradermal, intramuscular, and intraarticular administration.
- 60. The method of claim 55, wherein the recombinant adenovirus vector is administered to the liver.
- 61. The method of claim 60, wherein the recombinant adenovirus vector is delivered to the liver by a method selected from the group consisting of intravenous administration, intraportal administration, intrabiliary administration, intra-arterial administration, and direct injection into the liver parenchyma.
- 62. The method of claim 55, wherein the recombinant adenovirus vector is injected directly into a cancerous tissue.
- 63. The method of claim 55, wherein a therapeutic effect is achieved at a lower viral dose than with a non-replicating adenovirus vector.
- 64. A method of producing a polypeptide, comprising:
introducing the recombinant adenovirus vector of claim 19 into a plurality of cultured cells, under conditions sufficient for the recombinant adenovirus vector to be introduced into the cells and express the encoded polypeptide; collecting the polypeptide from the cell culture.
- 65. The method of claim 64, wherein the plurality of cultured cells are mammalian cells.
- 66. A method of producing an immune response against a pathogen in a subject, comprising:
administering to a subject a composition comprising a recombinant adenovirus vector according to claim 23 in a pharmaceutically acceptable carrier; wherein the composition is administered in an immunogenically effective amount and under conditions sufficient for the subject to produce an immune response against the pathogen.
- 67. A method of treating cancer, comprising
administering to a subject that has cancer a composition comprising an recombinant adenovirus vector according to claim 31 in a pharmaceutically acceptable carrier; wherein the composition is administered in an immunogenically effective amount and under conditions sufficient for the subject to produce an immune response against the cancer cell immunogen.
- 68. A method of treating cancer, comprising
administering to a subject that has cancer a composition comprising a recombinant adenovirus vector according to claim 33 in a pharmaceutically acceptable carrier, said heterologous nucleic acid encoding an anti-cancer agent; wherein the composition is administered in an immunogenically effective amount and under conditions sufficient for the subject to produce an immune response against the cancer cell immunogen.
- 69. A method of producing a recombinant adeno-associated virus (AAV) particle, comprising providing to a cell:
(a) a recombinant adenovirus vector according to claim 1;(b) a nucleic acid sequence encoding an AAV vector genome, said AAV vector genome comprising an AAV inverted terminal repeat sequence, a heterologous nucleic acid sequence, and an AAV packaging signal; (c) AAV rep coding sequences sufficient for replication of the AAV vector genome; (d) AAV cap coding sequences sufficient to produce a functional AAV capsid; wherein (a) to (d) are provided to the cell under conditions sufficient for replication and packaging of the AAV vector genome into the AAV capsid, whereby AAV particles comprising the AAV vector genome encapsidated within the AAV capsid are produced in the cell.
- 70. The method of claim 69, further comprising the step of collecting the recombinant AAV particles.
- 71. The method of claim 69, wherein the adenovirus vector genome comprises AAV cap sequences.
- 72. The method of claim 69, wherein the adenovirus vector genome comprises AAV rep sequences.
- 73. The method of claim 69, further comprising providing to the cell the adenovirus helper functions for AAV replication and packaging.
- 74. The method of claim 69, wherein the AAV inverted terminal repeats and the AAV capsid are derived from different AAV serotypes.
- 75. The method of claim 69, wherein the AAV capsid is an MV-6 capsid.
- 76. The method of claim 69, wherein the AAV inverted terminal repeats are AAV-2 inverted terminal repeats.
RELATED APPLICATION INFORMATION
[0001] This application claims the benefit of U.S. Provisional Application No. 60/295,914, filed Jun. 4, 2001, which is incorporated by reference herein in its entirety.
STATEMENT OF FEDERAL SUPPORT
[0002] The present invention was made, in part, with the support of grant number DK 52925 from the National Institutes of Health. The United States government has certain rights to this invention.
Provisional Applications (1)
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Number |
Date |
Country |
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60295914 |
Jun 2001 |
US |