Detection of cancer by elevated levels of BCL-2

• US 20080009005A1
• Filed: 02/09/2007
• Published: 01/10/2008
• Est. Priority Date: 02/09/2006
• Status: Active Grant

• Current Assignee: University of South Florida (State University System of Florida)
• Original Assignee: University of South Florida (State University System of Florida)
• Inventors: Kruk, Patricia
• Granted Patent: US 8,034,549 B2
• Time in Patent Office: Days
• US Class Current: 435/006
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/112 Disease subtyping, staging or classification
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 2333/47 Assays involving proteins of known structure or function as defined in the subgroups
• CPC Class Codes: G01N 33/57449 of ovaries
• CPC Class Codes: G01N 33/57484 involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

The present invention relates to a method for the diagnosis, prognosis, and monitoring of cancer, such as early or late stage ovarian cancer, in a subject by detecting Bcl-2 in a biological sample from the subject, preferably a urine or blood sample. Bcl-2 may be measured using an agent that detects or binds to Bcl-2 protein or an agent that detects or binds to encoding nucleic acids, such as antibodies specifically reactive with Bcl-2 protein or a portion thereof. The invention further relates to kits for carrying out the methods of the invention. The invention further relates to a device for the rapid detection of Bcl-2 in a bodily fluid and methods for rapidly measuring Bcl-2 in a bodily fluid.

Claims

1. A method of detecting cancer in a subject, comprising detecting the presence of Bcl-2 in a biological sample from the subject, wherein a level of Bcl-2 above a pre-determined threshold is indicative of cancer in the subject.

2. The method of claim 1, wherein said detecting comprises detecting Bcl-2 protein in the biological sample.

3. The method of claim 1, wherein said detecting comprises detecting a nucleic acid sequence encoding Bcl-2 protein in the biological sample.

4. The method of claim 1, wherein said detecting comprises:

(a) contacting the biological sample with a binding agent that binds Bcl-2 protein to form a complex;

(b) detecting the complex; and

correlating the detected complex to the amount of Bcl-2 protein in the sample, wherein the presence of elevated Bcl-2 protein is indicative of cancer.

5. The method of claim 1, wherein the biological sample is a biological fluid selected from the group consisting of urine, whole blood, serum, plasma, ascites fluid, and peritoneal fluid.

6. The method of claim 1, wherein the biological sample is urine.

7. The method of claim 1, wherein the cancer is selected from the group consisting of ovarian cancer, primary peritoneal cancer, and endometrial cancer.

8. The method of claim 1, wherein the cancer is ovarian cancer.

9. The method of claim 8, wherein the biological sample is urine or blood, and the cancer is ovarian cancer.

10. The method of claim 1, wherein the cancer is selected from the group consisting of breast cancer, endometrial cancer, cervical cancer, lung cancer, colon cancer, prostate cancer, melanoma, glioblastoma, sarcoma, bladder cancer, and head and neck cancer.

11. The method of claim 1, wherein the biological sample is urine or blood, and the cancer is prostate cancer.

12. The method of claim 4, wherein the binding agent is immobilized on a support.

13. The method of claim 4, wherein the binding agent is a monoclonal or polyclonal antibody.

14. The method of claim 4, wherein said detecting of (b) further comprises linking or incorporating a label onto the binding agent.

15. The method of claim 13, wherein said detecting of (b) uses ELISA-based immunoenzymatic detection.

16. The method of claim 1, further comprising detecting a biomarker of cancer in the same biological sample or a different biological sample obtained from the subject, before, during, or after said detecting of Bcl-2.

17. The method of claim 16, wherein the biomarker of cancer is a biomarker of gynecological cancer.

18. The method of claim 16, wherein the biomarker is CA125, LPA, or OVXI.

19. The method of claim 1, wherein said subject is suffering from cancer, and wherein said detecting is performed at several time points at intervals, as part of a monitoring of the subject before, during, or after the treatment of the cancer.

20. The method of claim 1, further comprising comparing the level of Bcl-2 in the biological sample with the level of Bcl-2 present in a normal control sample, wherein a higher level of Bcl-2 in the biological sample as compared to the level in the normal control sample is indicative of cancer.

21. The method of claim 1, wherein the subject is exhibiting no symptoms of cancer at the time said detecting is carried out.

22. The method of claim 1, wherein the subject is exhibiting one or more symptoms of cancer at the time said detecting is carried out.

23. The method of claim 1, wherein subject is exhibiting one or more of the symptoms selected from the group consisting of pelvic pain, abnormal vaginal bleeding, abdominal swelling or bloating, persistent back pain, persistent stomach upset, change in bowel or bladder pattern, pain during intercourse, unintentional weight loss of ten or more pounds, vulva or vaginal abnormality, change in the breast, and fatigue.

24. The method of claim 1, wherein the subject has an elevated CA125 level in the blood at the time of said detecting.

25. The method of claim 1, wherein the subject does not have an elevated CA125 level in the blood at the time of said detecting.

26. A method for prognostic evaluation of a subject having, or suspected of having, cancer, comprising:

a) determining the level of Bcl-2 in a biological sample obtained from the subject;

b) comparing the level determined in step (a) to a range of Bcl-2 known to be present in a biological sample obtained from a normal subject that does not have cancer; and

c) determining the prognosis of the subject based on the comparison of step (b), wherein a high level of Bcl-2 in step (a) indicates an aggressive form of cancer and, therefore, a poor prognosis.

27. A device for the rapid detection of Bcl-2 in a sample of bodily fluid, comprising an application zone for receiving a sample of bodily fluid;

a labeling zone containing a binding agent that binds to Bcl-2 in the sample; and

a detection zone where Bcl-2-bound binding agent is retained to give a signal, wherein the signal given for a sample from a subject with a Bcl-2 level lower than a threshold concentration is different from the signal given for a sample from a patient with a Bcl-2 level greater than a threshold concentration.

28. The device of claim 27, wherein the bodily fluid is urine and said threshold concentration is between 0 ng/ml and 2.0 ng/ml.

29. The device of claim 27, wherein the bodily fluid is urine and said threshold concentration is 1.8 ng/ml.

30. The device of claim 27, wherein the device has a reference zone which gives a signal which has the same intensity as the signal given in the detection zone for a sample from a subject having a Bcl-2 level equal to the threshold concentration.

31. The device of claim 27, wherein the binding agent is a tagged antibody.

32. The device of claim 31, wherein the antibody is tagged with colloidal gold.

33. The device of claim 27, wherein the detection zone comprises an immobilized anti-Bcl-2 antibody.

34. The device of claim 27, wherein the device has a control zone downstream of the detection zone which retains binding agent which passes through the detection zone.

35. The device of claim 27, wherein the control zone and the reference zone are the same zone.

36. The device of claim 27, wherein the binding agent is a tagged monoclonal antibody.

37. A method for measuring Bcl-2 in a bodily fluid, comprising:

(a) obtaining a sample of a bodily fluid from a subject;

(b) contacting the sample with a binding agent that binds to any Bcl-2 in the sample;

(c) separating Bcl-2-bound binding agent;

(d) detecting a signal associated with the separated binding agent from (c); and

(e) comparing the signal detected in step (d) with a reference signal which corresponds to the signal given by a sample from a subject with a Bcl-2 level equal to a threshold concentration.

38. The method of claim 37, wherein the bodily fluid is urine and said threshold concentration is between 0 ng/ml and 2.0 ng/ml.

39. The method of claim 37, wherein the bodily fluid is urine and said threshold concentration is between 1.8 ng/ml.

40. A kit for detection of cancer in a biological sample, comprising a binding agent specific for Bcl-2, and printed instructions for detecting cancer using in a biological fluid using the binding agent.

41. The kit of claim 40, further comprising one or more protease inhibitors to be applied to a biological sample.