METHOD FOR PREDICTING THE RISK OF GETTING CANCER OR DIAGNOSING CANCER IN A FEMALE SUBJECT

Abstract

Subject matter of the present invention is a method for predicting the risk of getting cancer in a female subject that does not suffer from cancer or alternatively diagnosing cancer in a female subject comprising: determining the level of Pro-Enkephalin or fragments thereof including Leu-Enkephalin and Met-Enkephalin of at least 5 amino acids in a bodily fluid obtained from said female subject; and correlating said level of Pro-Enkephalin or fragments thereof with a risk for getting cancer, wherein a reduced level is predictive for an enhanced risk of getting cancer or alternatively diagnosing cancer wherein an reduced level is correlated with the diagnosis of cancer.

Claims

1.-20. (canceled)

21. A method for predicting the risk of getting cancer in a female subject that does not suffer from cancer or, alternatively, diagnosing cancer in a female subject comprising: measuring the level of Pro-Enkephalin (SEQ ID No. 1), or one or more fragments thereof of at least 5 amino acids, in a sample of bodily fluid obtained from said female subject; and correlating said level of Pro-Enkephalin (SEQ ID No. 1), or of said one or more fragments thereof, with risk for getting cancer, wherein a reduced level of Pro-Enkephalin (SEQ ID No. 1), or said one or more fragments thereof is predictive for an enhanced risk of getting cancer or, alternatively, diagnosing cancer wherein a reduced level of Pro-Enkephalin (SEQ ID No. 1) or one or more fragments thereof is correlated with the diagnosis of cancer; wherein said one or more fragments thereof is selected from SEQ ID No. 2, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10 and SEQ ID No. 11.

22. The method according to claim 21, wherein said measuring is performed using an immunoassay that has antibodies or fragments of antibodies that bind to Pro-Enkephalin (SEQ ID No. 1) and/or said one or more fragments thereof.

23. The method according to claim 21, wherein said cancer is breast cancer or lung cancer.

24. The method according to claim 21, wherein said female subject has never had a history of diagnosis of cancer at the time said sample of bodily fluid is taken from said female subject.

25. The method according to claim 21, wherein said female subject has had a history of diagnosis of cancer and has been cured at the time said sample of bodily fluid is taken from said female subject and the risk of reoccurrence of getting breast cancer is determined or alternatively the reoccurrence of breast cancer is diagnosed.

26. The method according to claim 21, wherein at the time said sample of bodily fluid is taken from said female subject, said female subject has been diagnosed as having a cardiovascular disease or diabetes.

27. The method according to claim 21, further comprising determining at least one clinical parameter selected from: age, presence of diabetes mellitus, and currently smoking.

28. The method according to claim 21, wherein the level of Pro-Enkephalin (SEQ ID No. 1) is measured with an immunoassay.

29. The method according to claim 21, wherein said method is performed more than once in order to monitor the risk of getting breast cancer in said female subject or in order to monitor a course of treatment following a diagnosis of breast cancer.

30. The method according to claim 21, wherein said a method is performed to monitor the response of said female subject to preventive and/or therapeutic measures taken following an assessment of the risk of getting cancer or following a diagnosis of cancer.

31. The method according to claim 21, wherein said a method is performed in order to stratify female subjects into risk groups.

32. The method according to claim 21, wherein said method further comprises: measuring the level of Pro-Neurotensin 1-117 (SEQ ID No. 17) in a sample of bodily fluid obtained from said female subject; and correlating said level of Pro-Neurotensin 1-117 (SEQ ID No. 17) with risk for getting cancer, wherein an increased level of Pro-Neurotensin 1-117 (SEQ ID No. 17) is predictive for an enhanced risk of getting cancer, or, alternatively, diagnosing cancer wherein an increased level of Pro-Neurotensin 1-117 (SEQ ID No. 17) is correlated with the diagnosis of cancer.

33. The method according to claim 21, wherein said reduced level of Pro-Enkephalin (SEQ ID No. 1) or of said one or more fragments thereof is a level below 100 pmol/l.

34. The method according to claim 32, wherein said reduced level of Pro-Enkephalin (SEQ ID No. 1) or of said one or more fragments thereof is a level below 100 pmol/l.

35. The method according to claim 32, wherein said increased level of Pro-Neurotensin 1-117 (SEQ ID No. 17) is a level above 78 pmol/l.

36. The method according to claim 34, wherein said increased level of Pro-Neurotensin 1-117 (SEQ ID No. 17) is a level above 78 pmol/l.

37. The method according to claim 21, wherein said bodily fluid is blood, plasma or serum.

38. The method according to claim 21, wherein the level of a fragment of Pro-Enkephalin (SEQ ID No. 1) is measured and said fragment is MR-Pro-Enkephalin (SEQ ID No. 6).

39. A kit for determining Pro-Enkephalin (SEQ ID No. 1) and/or one or more fragments thereof in a sample comprising a first and second binder, wherein the first binder binds to a region of Pro-Enkephalin (SEQ ID No. 1) that is within amino acid sequence 133-140 (LKELLETG, SEQ ID NO. 22), and the second binder binds to a region of Pro-Enkephalin (SEQ ID No. 1) that is within amino acid sequence 152-159 (SDNEEEVS, SEQ ID NO. 23), wherein each of said regions comprises at least 4 or 5 amino acids.

40. The kit according to claim 39, wherein said kit is sufficiently sensitive to quantify the level of Pro-Enkephalin (SEQ ID No. 1) or said one or more fragments thereof of healthy subjects and has an analytical assay sensitivity<15pmol/L.

41. A method for predicting a risk of getting cancer in a female subject that does not suffer from cancer or, alternatively, diagnosing cancer in a female subject, and optionally for stratification of female subjects into risk groups, said method comprising: measuring the level of Pro-Enkephalin (SEQ ID No. 1), or of one or more fragments thereof, in a sample of bodily fluid obtained from a female subject using said kit according to claim 39; correlating said level of Pro-Enkephalin, or of one or more fragments thereof, with risk for getting cancer, wherein a reduced level of Pro-Enkephalin (SEQ ID No. 1), or said one or more fragments thereof is predictive for an enhanced risk of getting cancer or, alternatively, diagnosing cancer wherein a reduced level of Pro-Enkephalin (SEQ ID No. 1) or one or more fragments thereof is correlated with the diagnosis of cancer; and optionally using said method to stratify female subjects into risk groups.

42. The method according to claim 41, wherein said method is used for stratification of female subjects into a group that should obtain opioid growth factor (OGF) therapy.

43. The method according to claim 41, wherein said method is used for predicting the risk of getting cancer in said female subject that does not suffer from cancer.

44. The method according to claim 22, wherein said cancer is breast cancer.

45. The method according to claim 32, wherein said cancer is breast cancer.

46. The method according to claim 21,wherein said correlating is performed by: (a) comparing the level of Pro-Enkephalin (SEQ ID No. 1) or of said one or more fragments thereof in said sample with the median of the level of Pro-Enkephalin (SEQ ID No. 1) or of said one or more fragments thereof in an ensemble of pre-determined samples in a population of healthy or apparently healthy subjects, (b) comparing the level of Pro-Enkephalin (SEQ ID No. 1) or of said one or more fragments thereof in said sample with a quantile of the level of Pro-Enkephalin (SEQ ID No. 1) or of said one or more fragments thereof in an ensemble of pre-determined samples in a population of healthy or apparently healthy subjects, or (c) calculating the risk by Cox Proportional Hazards analysis or by Risk index calculations using the level of Pro-Enkephalin (SEQ ID No. 1) or of said one or more fragments thereof in said sample.

47. A method according to claim 21, wherein said measuring of the level of Pro-Enkephalin (SEQ ID No. 1), or of said one or more fragments thereof, comprises: a) bringing said sample of bodily fluid into contact with a solid phase comprising a bound first antibody or first antibody fragment that binds to Pro-Enkephalin (SEQ ID No. 1) or to said one or more fragments thereof whereby Pro-Enkephalin or said one or more fragments thereof within said sample react with said bound first antibody or antibody fragment to form a complex bound to said solid phase, b) contacting said solid phase with the bound complex with a second antibody or second antibody fragment, wherein said second antibody or second antibody fragment is labelled with a detectable label, and whereby the labelled second antibody or second antibody fragment binds to said complex, and c) measuring the level of Pro-Enkephalin (SEQ ID No. 1) or of said one or more fragments thereof in said sample by measuring the amount of labelled second antibody or second antibody fragment bound to the complex on said solid phase.

48. A method according to claim 47, wherein one of either (a) said bound first antibody or first antibody fragment, or (b) said labelled second antibody or second antibody fragment, binds to a peptide consisting of amino acid sequence 133-140 of Pro-Enkephalin (SEQ ID No. 1), and the other of (a) said bound first antibody or first antibody fragment, and (b) said labelled second antibody or second antibody fragment, binds to a peptide consisting of amino acid sequence 152-159 of Pro-Enkephalin (SEQ ID No. 1).

49. A method according to claim 32, wherein said measuring of the level of Pro-Enkephalin (SEQ ID No. 1), or of said one or more fragments thereof, comprises: a) bringing said sample of bodily fluid into contact with a solid phase comprising a bound first antibody or first antibody fragment that binds to Pro-Enkephalin (SEQ ID No. 1) or to said one or more fragments thereof whereby Pro-Enkephalin or said one or more fragments thereof within said sample react with said bound first antibody or antibody fragment to form a complex bound to said solid phase, b) contacting said solid phase with the bound complex with a second antibody or second antibody fragment, wherein said second antibody or second antibody fragment is labelled with a detectable label, and whereby the labelled second antibody or second antibody fragment binds to said complex, and c) measuring the level of Pro-Enkephalin (SEQ ID No. 1) or of said one or more fragments thereof in said sample by measuring the amount of labelled second antibody or second antibody fragment bound to the complex on said solid phase.

50. A method according to claim 49, wherein one of either (a) said bound first antibody or first antibody fragment, or (b) said labelled second antibody or second antibody fragment, binds to a peptide consisting of amino acid sequence 133-140 of Pro-Enkephalin (SEQ ID No. 1), and the other of (a) said bound first antibody or first antibody fragment, and (b) said labelled second antibody or second antibody fragment, binds to a peptide consisting of amino acid sequence 152-159 of Pro-Enkephalin (SEQ ID No. 1).

51. A method according to claim 49, wherein said measuring of the level of Pro-Neurotensin 1-117 (SEQ ID No. 17) comprises: a) bringing said sample of bodily fluid into contact with a solid phase comprising a bound third antibody or third antibody fragment that binds to Pro-Neurotensin 1-117 (SEQ ID No. 17) whereby Pro-Neurotensin 1-117 (SEQ ID No. 17) within said sample reacts with said bound third antibody or third antibody fragment to form a complex bound to said solid phase, b) bringing said solid phase with the bound complex into contact with a fourth antibody or fourth antibody fragment, wherein said fourth antibody or fourth antibody fragment is labelled with a detectable label, and whereby the labelled fourth antibody or fourth antibody fragment binds to said complex, c) measuring the level of Pro-Neurotensin 1-117 (SEQ ID No. 17) in said sample by measuring the amount of labelled fourth antibody or fourth antibody fragment bound to the complex on said solid phase.

52. A method according to claim 51, wherein one of either (a) said bound third antibody or third antibody fragment, or (b) said labelled fourth antibody or fourth antibody fragment, binds to a peptide consisting of amino acid sequence 1-19 of Pro-Neurotensin 1-117 (SEQ ID No. 17), and the other of (a) said bound third antibody or third antibody fragment, and (b) said labelled fourth antibody or fourth antibody fragment, binds to a peptide consisting of amino acid sequence 44-62 of Pro-Neurotensin 1-117 (SEQ ID No. 17).

53. A method according to claim 52, wherein one of either (a) said bound first antibody or first antibody fragment, or (b) said labelled second antibody or second antibody fragment, binds to a peptide consisting of amino acid sequence 133-140 of Pro-Enkephalin (SEQ ID No. 1), and the other of (a) said bound first antibody or first antibody fragment, and (b) said labelled second antibody or second antibody fragment, binds to a peptide consisting of amino acid sequence 152-159 of Pro-Enkephalin (SEQ ID No. 1).

Description

FIGURE DESCRIPTION

[0124] FIG. 1: shows a typical Pro-Enkephalin dose/signal curve. Standard curve Pro-Enkephalin.

[0125] FIG. 2: frequence distribution of Pro-Enkephalin in the females population:

[0126] FIG. 3: Kaplan Meier graphs, illustrating the cumulative breast cancer diagnosis in women quartile (Q) 1 (below 40.4 pmol/l) quartile 2 (40.4-47.1 pmol/l), quartile 3 (47.2-54.1 pmol/l), quartile 4 (above 54.1 pmol/l). Decreased PENK indicates a long term increased risk of breast cancer development. Since any women with cancer history at day of baseline (blood sampling) were excluded, Pro-Enkephalin is highly predictive for future breast cancer development. Over all, women from Q 1 have a 3.6 times higher risk to develop breast cancer than women from Q 4.

[0127] FIG. 4: Illustration example of combined analysis of Pro-Enkephalin for breast cancer prediction: