Adenovirus 36 E4 orf 1 Gene and Protein and Their Uses

• US 20090264356A1
• Filed: 11/30/2006
• Published: 10/22/2009
• Est. Priority Date: 11/30/2005
• Status: Active Grant

• Current Assignee: Board of Supervisors of Louisiana State University Agricultural & Mechanical College (Louisiana State University System), Wayne State University
• Original Assignee: Wayne State University
• Inventors: Dhurandhar, Nikhil V., Wang, Zhong Q., Holland, Thomas C.
• Granted Patent: US 8,008,436 B2
• Time in Patent Office: Days
• US Class Current: 514/1.100
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 2710/10322 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2710/10332 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent

Abstract

Expression of the E4 orf 1 gene of Ad-36 alone has been discovered to be responsible for the increased insulin sensitivity observed in Ad-36 infected animals, including increased adipogenesis. Ad-36 E4 orf 1 protein can be used to increase insulin sensitivity and ameliorate diabetes. Additionally, drugs that mimic the action of Ad-36 E4 orf 1 protein could be found. Ad-36 E4 orf 1 could also be used to increase fat cells in lipodystrophy. We have also discovered that Ad-36 infection in human skeletal muscle cells increased differentiation and insulin independent glucose uptake. It is expected that infection with Ad-36 E4 orf 1 gene will also cause these effects.

Claims

1. An isolated nucleic acid sequence comprising SEQ ID NO:

1, or comprising oligonucleotides that encodes the same amino acid sequence in accordance with the degeneracy of the genetic code.

2. The nucleic acid recited in claim 1, wherein said nucleic acid comprises SEQ ID NO:

1.

3. The nucleic acid recited in claim 1, wherein said nucleic acid is SEQ ID NO:

1.

4. A transformation vector comprising the nucleic acid sequence recited in claim 1.

5. A nucleic acid sequence as recited in claim 1, wherein said sequence is operably linked to a promoter that is functional in animal cells.

6. An animal cell transformed with the nucleic acid sequence recited in claim 5.

17. A method for enhancing or inducing adipogenesis in a mammalian patient, comprising administering to the patient'"'"'s adipose tissue a vector as recited in claim 4;

wherein expression of the encoded amino acid sequence in the patient'"'"'s adipose tissue results in adipogenesis.

18. The method of claim 17, wherein the nucleic acid sequence comprises SEQ ID NO:

1.

19. A method for enhancing or inducing glucose uptake in a mammalian patient, comprising administering to the patient'"'"'s skeletal muscle tissue a vector as recited in claim 4;

wherein expression of the encoded amino acid sequence in the patient'"'"'s skeletal muscle tissue results in increased glucose uptake.

20. The method of claim 19, wherein the nucleic acid sequence comprises SEQ ID NO:

1.

21. A method for enhancing or inducing insulin sensitivity in a mammalian patient, comprising administering to the patient'"'"'s adipose tissue a vector as recited in claim 4;

wherein expression of the encoded amino acid sequence in the patient'"'"'s adipose tissue results in increased insulin sensitivity.

22. The method of claim 21, wherein the nucleic acid sequence comprises SEQ ID NO:

1.

23. The transformation vector of claim 4, wherein the vector is adapted to transpose mammalian adipose cells or mammalian skeletal muscle cells.

7. An isolated protein whose amino acid sequence comprises SEQ ID NO:

2.

8. The protein recited in claim 7, wherein the amino acid sequence of said protein is SEQ ID NO:

2.

9. A method of treating or ameliorating the symptoms of a disease in a mammalian patient wherein the disease is chosen from the group consisting of lipodystrophy, diabetes, and insulin resistance, said method comprising administering to the patient a therapeutically effective amount of Adenovirus-36 E4 orf 1 protein, wherein the patient'"'"'s symptoms improve following said administering.

10. A method as in claim 9, wherein the disease is insulin resistance, and wherein the patient'"'"'s insulin sensitivity increases following said administering.

11. A method as in claim 9, wherein the disease is lipodystrophy and wherein the number of adipose tissue cells in the patient increases following said administering.

12. A method as in claim 9, wherein the disease is diabetes due to a concentration of high serum glucose, and wherein the amount of serum glucose decreases following said administering.

13. The method of claim 9, wherein the amino acid sequence of the Adenovirus-36 E4 orf 1 protein is SEQ ID NO:

2.

14. A method as in claim 9, wherein the Adenovirus-36 E4 orf 1 protein is administered by introducing into the mammal a nucleic acid sequence encoding the Adenovirus-36 E4 orf 1 protein, in a manner permitting expression of the Adenovirus-36 E4 orf 1 protein.

15. A method of claim 14, wherein the nucleic acid sequence is introduced by a method selected from group consisting of electroporation, DEAE Dextran transfection, calcium phosphate transfection, cationic liposome fusion, proptoplast fusion, creation of an in vivo electric field, DNA-coated microprojectile bombardment, injection with recombinant replication-defective viruses, homologous recombination, in vivo gene therapy, ex vivo gene therapy, viral vectors, and naked DNA transfer.

16. A method as in claim 14, wherein the nucleic acid sequence comprises SEQ ID NO:

1.

24. The method of claim 9, wherein the Adenovirus-36 E4 orf 1 protein is selected from the group consisting of an analog and derivative of a protein with the amino acid sequence of SEQ ID NO:

2.

25. A method for predicting insulin sensitivity in a patient, said method comprising assaying a tissue from the patient for the presence of one or more compounds selected from the group consisting of Adenovirus-36 E4 orf 1 DNA, Adenovirus-36 E4 orf 1 mRNA, and Adenovirus-36 E4 orf 1 protein, wherein the presence of said compound indicates a patient with a higher insulin sensitivity than a patient without said compound.

26. The method of claim 25, wherein the assayed tissue is one or more tissues from the group consisting of adipose tissue, blood, and other body tissues and fluids.

27. The method of claim 25, wherein the Adenovirus-36 E4 orf 1 DNA has the nucleic acid sequence of SEQ ID NO:

1.

28. The method of claim 25, wherein the Adenovirus-36 E4 orf 1 protein has the amino acid sequence of SEQ ID NO:

2.

29. The method of claim 25, wherein the assay for the presence of said compound is one or more assays selected from the group consisting of real time PCR, reverse transcription PCR, Northern blot analysis, enzyme linked immunosorbet assay, or Western Blot.

30. The method of claim 25, wherein the patient is substantially overweight.

31. The method of claim 25, wherein the patient is diabetic.