Diagnosis and Treatment of Chronic Lymphocytic Leukemia (CLL)

• US 20090131353A1
• Filed: 12/07/2006
• Published: 05/21/2009
• Est. Priority Date: 12/07/2005
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Zhang, Lingzhi, Insel, Paul A., Kipps, Thomas, Kanter, Joan R., Rassenti, Laura, Murray, Fiona
• Granted Patent: US 7,846,664 B2
• Time in Patent Office: Days
• US Class Current: 514/44.R
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/158 Expression markers

Abstract

The present invention provides diagnostic methods and kits for diagnosis of chronic lymphocytic leukemia (CLL) by determining expression levels of isoforms of cyclic nucleotide phosphodiesterases (PDEs) associated with CLL particularly, PDE7B and/or PDE3B, and a ratio of mRNA expression of PDE7B to PDE3B. The present invention provides that CLL lymphocytes uniformly expressed high levels of PDE7B and low levels of PDE3B relative to those of normal lymphocytes. A method of treatment and a pharmaceutical composition for CLL comprising one or more therapeutic agents capable of modulating expression or activity levels of isoforms of PDEs associated with CLL, and/or reversing the ratio of PDE7B/PDE3B mRNA expression levels are also provided.

Claims

1. A diagnostic method for chronic lymphocytic leukemia (CLL) comprising:

(a) determining in a sample from a patient an expression level of an isoform of phosphodiesterase (PDE) associated with CLL,

(b) comparing the patient'"'"'s expression level to the expression level of the isoform of PDE associated with CLL in a normal subject, and

(c) diagnosing said patient as at risk for CLL when the patient'"'"'s expression level is significantly increased or decreased compared to said normal subject'"'"'s expression level.

2. The diagnostic method of claim 1, wherein said isoform of PDE is phosphodiesterase 7B (PDE7B), wherein PDE7B expression level in said patient is significantly increased, as compared to that in said normal subject.

3. The diagnostic method of claim 2, wherein said PDE7B expression level comprises PDE7B mRNA expression level or PDE7B protein expression level.

4. The diagnostic method of claim 3, wherein said PDE7B mRNA expression level in said patient is about 5˜

70-fold increased, as compared to that in said normal subject.

5. The diagnostic method of claim 3, wherein said PDE7B protein expression level in said patient is about 5˜

90-fold increased, as compared to that in said normal subject.

6. The diagnostic method of claim 1, wherein said isoform of PDE is phosphodiesterase 3B (PDE3B), wherein PDE3B mRNA expression level in said patient is significantly decreased, as compared to that in normal lymphocytes.

7. The diagnostic method of claim 6, wherein said PDE3B expression level in said patient is about 20-fold decreased, as compared to that in normal lymphocytes.

8. The diagnostic method of claim 1 further comprising determining a ratio of phosphodiesterase 7B (PDE7B) and phosphodiesterase 3B (PDE3B) mRNA expression levels, wherein said ratio of PDE7B/PDE3B mRNA expression levels in said patient is significantly increased, as compared to that in said normal subject.

9. The diagnostic method of claim 8, wherein an average ratio of PDE7B/PDE3B mRNA expression levels in said patient is about 1000-fold increased, as compared to that in said normal subject.

10. The diagnostic method of claim 1, wherein said sample comprises lymphocytes comprising peripheral blood mononuclear cells (PBMCs) or B cells.

11. A kit for a diagnosis of CLL comprising biomarker detecting reagents for determining a differential expression level of isoforms of PDE, and instructions for their use in diagnosing CLL.

12. The kit of claim 11, wherein said isoform of PDE is PDE7B, and wherein both PDE7B mRNA and protein expression levels in a sample from a patient as at risk for CLL is significantly increased compared to that in a normal subject.

13. The kit of claim 12, wherein said PDE7B mRNA expression level in said patient as at risk for CLL is about 5-70 fold increased, as compared to that in a normal subject.

14. The kit of claim 12, wherein said PDE7B protein expression level in said patient as at risk for CLL is about 5-90 fold increased, as compared to that in a normal subject.

15. The kit of claim 11, wherein said isoform of PDE is PDE3B, and wherein PDE3B mRNA expression level in a sample from a patient as at risk for CLL is significantly decreased compared to that in a normal subject.

16. The kit of claim 15, wherein said PDE3B mRNA expression level in said patient as at risk for CLL is about 20 fold decreased compared to that in a normal subject.

17. The kit of claim 11, further comprising means for determining a ratio of PDE7B and PDE3B mRNA expression levels in a patient as at risk for CLL, wherein said ratio of PDE7B/PDE3B mRNA expression levels in said patient is significantly increased compared to that in a normal subject.

18. The kit of claim 17, wherein an average ratio of PDE7B/PDE3B mRNA expression levels in said patients is about 1000-fold increased, as compared to that in said normal subjects.

19. A method of a treatment comprising administering to a patient in need thereof a biologically effective amount of one or more therapeutic agents capable of directly or indirectly modulating expression or activity levels of one or more isoforms of PDE associated with CLL.

20. The method of claim 19, wherein one of said isoforms of PDE comprises PDE7B.

21. The method of claim 20, wherein said therapeutic agents are capable of directly or indirectly inhibiting PDE7B mRNA expression, PDE7B protein expression or enzyme activity in CLL lymphocytes.

22. The method of claim 21, wherein said therapeutic agents comprises oligonucleotides, antisense oligonucleotides, polynucleotides, therapeutic DNA, ribozymes, dsRNAs, siRNA, RNAi, or gene therapy vectors.

23. The method of claim 21, wherein said therapeutic agents comprise PDE7B inhibitors, antagonists, antibodies or other immunologically active molecules that immunoreact with PDE7B protein, resulting in apoptosis of CLL lymphocytes.

24. The method of claim 20, wherein said therapeutic agents are capable of reversing a ratio of PDE7B/PDE3B mRNA expression levels in CLL lymphocytes.

25. The method of claim 24, wherein said CLL lymphocytes comprise peripheral blood mononuclear cells (PBMCs) or B cells.

26. A pharmaceutical composition for treating CLL comprising an effective amount of one or more therapeutic agents capable of directly or indirectly modulating expression or activity levels of one or more isoforms of PDE associated with CLL.

27. The pharmaceutical composition of claim 26, wherein one of said isoforms of PDE comprises PDE7B.

28. The pharmaceutical composition of claim 27, wherein, said therapeutic agents are capable of directly or indirectly inhibiting PDE7B mRNA expression, PDE7B protein expression, or enzyme activity in CLL lymphocytes.

29. The pharmaceutical composition of claim 28, wherein said therapeutic agents comprises oligonucleotides, antisense oligonucleotides, polynucleotides, therapeutic DNA, ribozymes, dsRNAs, siRNA, RNAi, or gene therapy vectors.

30. The pharmaceutical composition of claim 29, wherein said therapeutic agents comprise PDE7B inhibitors, antagonists, antibodies or other immunologically active molecules that immunoreact with PDE7B protein, resulting in apoptosis of CLL lymphocytes.

31. The pharmaceutical composition of claim 26, wherein said therapeutic agents are capable of reversing a ratio of PDE7B/PDE3B mRNA expression levels in CLL lymphocytes.

32. The pharmaceutical composition of claim 31, wherein said CLL lymphocytes comprise peripheral blood mononuclear cells (PBMCs) or B cells.