Use of resveratrol to regulate expression of apolipoprotein A1

• US 20060147904A1
• Filed: 03/15/2006
• Published: 07/06/2006
• Est. Priority Date: 08/15/2002
• Status: Abandoned Application

• Current Assignee: Resverlogix Corp.
• Original Assignee: Resverlogix Corp.
• Inventors: Wong, Norman C.W.
• Application Number:
  US11/375,628
• Publication Number:
  US 20060147904A1
• Time in Patent Office: Days
• US Class Current: 435/4
• CPC Class Codes: A61K 31/05 Phenols cannabinoids A61K31/658
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 1/6897 involving reporter genes operably linked to promoters
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 2500/10 involving cells
• CPC Class Codes: G01N 2800/044 Hyperlipemia or hypolipemia, e.g. dyslipidaemia, obesity
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743

Abstract

Described are new methods for promoting the expression of apolipoprotein A1 (APO A1) for increasing levels of HDL, and assays for screening and identifying compounds for regulating expression of the APO A1 protein.

Claims

1-13. -13. (canceled)

14. A method for screening and identifying a compound that is capable of increasing plasma HDL levels in a mammal, comprising:

a) exposing cells to the compound to be screened; and

b) determining whether the compound acts upon the −

190 to −

170 DNA motif of the APO AI gene, wherein the +1 position is the transcription start of the gene, thereby increasing APO AI gene expression,

thereby identifying a compound capable of increasing plasma HDL levels.

15. The method of claim 14, wherein the compound is a resveratrol analog and/or mimetic.

16. The method of claim 14, wherein the cells are Hep G2 cells.

17. The method of claim 14, wherein the cells are CaCO2 cells.

18. The method of claim 14, wherein the cells are permanently transfected with a plasmid comprising a promoter sequence comprising the −

190 to −

170 DNA motif of the APO AI gene operably linked to a reporter gene.

19. The method of claim 18, wherein the cells are Hep G2 cells.

20. The method of claim 18, wherein the cells are CaCO2 cells.

21. The method of claim 18, wherein the promoter sequence is a full or truncated APO AI gene promoter sequence.

22. The method of claim 18, wherein the plasmid is pAI.474.

23. The method of claim 18, wherein the reporter gene is Luciferase.

24. The method of claim 18, wherein APO AI gene expression activity is measured at least about 16 hours after exposing the cells to the compound.

25. The method of claim 14, comprising measuring APO AI gene expression activity by assaying for levels of Apo AI protein.

26. The method of claim 25, wherein assaying for levels of Apo AI protein comprises using Western blotting.

27. The method of claim 25, wherein the levels of Apo AI protein are measured at least about 36 hours after exposing the cells to the compound.

28. The method of claim 18, further comprising exposing a separate population of the cells to the compound.

29. The method of claim 28, further comprising measuring APO A1 gene expression activity by assaying the separate population of the cells for levels of Apo AI protein.

30. The method of claim 29, wherein assaying for levels of Apo Al protein comprises using Western blotting.