BIOMARKER FOR DIAGNOSIS, PREDICTION AND/OR PROGNOSIS OF ACUTE HEART FAILURE AND USES THEREOF

1 . A method for diagnosing, predicting and/or prognosticating acute heart failure (AHF) in a subject, characterised in that the examination phase of the method comprises measuring the quantity of MCAM in a sample from the subject.

2 . The method according to claim 1 comprising the steps:

(i) measuring the quantity of MCAM in the sample from the subject;

(ii) comparing the quantity of MCAM measured in (i) with a reference value of the quantity of MCAM, said reference value representing a known diagnosis, prediction and/or prognosis of AHF;

(iii) finding a deviation or no deviation of the quantity of MCAM measured in (i) from the reference value;

(iv) attributing said finding of deviation or no deviation to a particular diagnosis, prediction and/or prognosis of AHF in the subject.

3 . The method according to claim 1 or 2 , wherein an elevated quantity of MCAM in the sample from the subject compared to a reference value representing the diagnosis or prediction of no AHF or representing a good prognosis for AHF indicates that the subject has or is at risk of having AHF or indicates a poor prognosis for AHF in the subject.

4 . The method according to any one of claims 1 to 3 , wherein the sensitivity and/or specificity of the method is at least 80%.

5 . The method according to any one of claims 1 to 4 , wherein the subject presents itself with one or more symptoms and/or signs potentially indicative of AHF.

6 . The method according to any one of claims 1 to 5 , wherein the subject presents itself with dyspnea, e.g. caused by COPD, pneumonia or AHF.

7 . The method according to any one of claims 1 to 6 , wherein the subject has a medical history of heart failure.

8 . A method for diagnosing acute heart failure (AHF) or recovery thereof in a subject admitted to the hospital comprising the steps:

(i) measuring the quantity of MCAM in the sample from the subject upon admission to the hospital and at a time point where a diagnosis of recovery of AHF has to be made;

(ii) comparing the quantity of MCAM measured in (i) with a reference value of the quantity of MCAM, said reference value representing a known diagnosis, prediction and/or prognosis of AHF;

(iii) finding a deviation or no deviation of the quantity of MCAM measured in (i) from the reference value;

(iv) attributing said finding of deviation or no deviation to a particular diagnosis of AHF or recovery of AHF in the subject.

9 . A method for monitoring a change in the diagnosis, prediction and/or prognosis of AHF in a subject, comprising:

(i) applying the method of any one of claims 1 to 8 to the subject at one or more successive time points, whereby the diagnosis, prediction and/or prognosis of AHF in the subject is determined at said successive time points;

(ii) comparing the diagnosis, prediction and/or prognosis of AHF in the subject at said successive time points as determined in (i); and

(iii) finding the presence or absence of a change between the diagnosis, prediction and/or prognosis of AHF in the subject at said successive time points as determined in (i).

10 . The method according to claim 9 , wherein said change in the diagnosis, prediction and/or prognosis of AHF in the subject is monitored in the course of a medical treatment of said subject.

11 . The method according to any one of claims 1 to 10 , wherein the examination phase of the method further comprises measuring the presence or absence and/or quantity of one or more other biomarkers useful for diagnosing, predicting and/or prognosticating AHF in the sample from the subject.

12 . The method according to claim 11 comprising the steps:

(i) measuring the quantity of MCAM and the presence or absence and/or quantity of said one or more other biomarkers in the sample from the subject;

(ii) using the measurements of (i) to establish a subject profile of the quantity of MCAM and the presence or absence and/or quantity of said one or more other biomarkers;

(iii) comparing said subject profile of (ii) to a reference profile of the quantity of MCAM and the presence or absence and/or quantity of said one or more other biomarkers, said reference profile representing a known diagnosis, prediction and/or prognosis of AHF;

(iv) finding a deviation or no deviation of the subject profile of (ii) from the reference profile;

(v) attributing said finding of deviation or no deviation to a particular diagnosis, prediction and/or prognosis of AHF in the subject.

13 . The method according to any one of claim 11 or 12 , wherein said other biomarker useful for diagnosing, predicting and/or prognosticating AHF is chosen from the group consisting of B-type natriuretic peptide (BNP), pro-B-type natriuretic peptide (proBNP), amino terminal pro-B-type natriuretic peptide (NTproBNP), and fragments of any one thereof.

14 . A method for establishing a reference value for the quantity of MCAM, said reference value representing:

(a) a diagnosis or prediction of no AHF or a good prognosis for AHF, or

(b) a diagnosis or prediction of AHF or a poor prognosis for AHF, comprising:

(i) measuring the quantity of MCAM in:

(i a) one or more samples from one or more subjects not having AHF or not being at risk of having AHF or having a good prognosis for AHF, or

(i b) one or more samples from one or more subjects having AHF or being at risk of having AHF or having a poor prognosis for AHF, and

(ii) storing the quantity of MCAM

(ii a) as measured in (i a) as the reference value representing the diagnosis or prediction of no AHF or representing the good prognosis for AHF, or

(ii b) as measured in (i b) as the reference value representing the diagnosis or prediction of AHF or representing the poor prognosis for AHF.

15 . A method for establishing a reference profile for the quantity of MCAM and the presence or absence and/or quantity of one or more other biomarkers useful for diagnosing, predicting and/or prognosticating AHF, said reference profile representing:

(a) a diagnosis or prediction of no AHF or a good prognosis for AHF, or

(b) a diagnosis or prediction of AHF or a poor prognosis for AHF, comprising:

(i) measuring the quantity of MCAM and the presence or absence and/or quantity of said one or more other biomarkers in:

(i a) one or more samples from one or more subjects not having AHF or not being at risk of having AHF or having a good prognosis for AHF; or

(i b) one or more samples from one or more subjects having AHF or being at risk of having AHF or having a poor prognosis for AHF;

(ii)

(ii a) using the measurements of (i a) to create a profile of the quantity of MCAM and the presence or absence and/or quantity of said one or more other biomarkers; or

(ii b) using the measurements of (i b) to create a profile of the quantity of MCAM and the presence or absence and/or quantity of said one or more other biomarkers;

(iii)

(iii a) storing the profile of (ii a) as the reference profile representing the diagnosis or prediction of no AHF or representing the good prognosis for AHF; or

(iii b) storing the profile of (ii b) as the reference profile representing the diagnosis or prediction of AHF or representing the poor prognosis for AHF.

16 . The method according to claim 15 , wherein said other biomarker useful for diagnosing, predicting and/or prognosticating AHF is chosen from the group consisting of B-type natriuretic peptide (BNP), pro-B-type natriuretic peptide (proBNP), amino terminal pro-B-type natriuretic peptide (NTproBNP), and fragments of any one thereof.

17 . A method for establishing a MCAM base-line or reference value in a subject, comprising:

(i) measuring the quantity of MCAM in the sample from the subject at different time points wherein the subject is not suffering from AHF, and

(ii) calculating the range or mean value of the subject, which is the MCAM base-line or reference value for said subject.

18 . The method according to any one of claims 1 to 17 , wherein the subject is human.

19 . The method according to claim 18 , wherein the subject is suffering from AHF involving systolic dysfunction.

20 . The method according to claim 19 , wherein said systolic dysfunction is characterized by a decreased left ventricular ejection fraction (LVEF), preferably wherein said LVEF is less than 55% or less than 50% or less than 45%, and/or by increased cardiac filling pressure.

21 . The method according to any one of claims 1 to 20 , wherein the quantity of MCAM and/or the presence or absence and/or quantity of the one or more other biomarkers is measured using, respectively, a binding agent capable of specifically binding to MCAM and/or to fragments thereof, and a binding agent capable of specifically binding to said one or more other biomarkers.

22 . The method according to any one of claims 1 to 20 , wherein the quantity of MCAM and/or the presence or absence and/or quantity of the one or more other biomarkers is measured using an immunoassay technology, such as direct ELISA, indirect ELISA, sandwich ELISA, competitive ELISA, multiplex ELISA, radioimmunoassay (RIA) or ELISPOT technologies, or using a mass spectrometry analysis method or using a chromatography method, or using a combination of said methods.

23 . The method of any one of claims 1 - 11 , wherein the reference value is determined according to any one of claims 13 - 22 .

24 . A reference value obtained according to any one of claims 14 - 22 .

25 . The method according to any one of claims 1 - 22 , wherein the sample used is plasma, and wherein the plasma circulating form of MCAM is detected.

26 . A kit for diagnosing, predicting and/or prognosticating AHF in a subject, the kit comprising means for measuring the quantity of MCAM in a sample from the subject.

27 . The kit according to claim 26 , comprising:

(i) means for measuring the quantity of MCAM in the sample from the subject; and

(ii) a reference value of the quantity of MCAM or means for establishing said reference value, wherein said reference value represents a known diagnosis, prediction and/or prognosis of AHF.

28 . The kit according to any one of claim 26 or 27 , further comprising means for measuring in the sample from the subject the presence or absence and/or quantity of one or more other biomarkers useful for diagnosing, predicting and/or prognosticating AHF, preferably wherein said other biomarkers are chosen from the group consisting of BNP, proBNP, NTproBNP and fragments of any one thereof.

29 . The kit according to claim 28 , comprising:

(i) means for measuring the quantity of MCAM in the sample from the subject;

(ii) means for measuring the presence or absence and/or quantity of said one or more other biomarkers in the sample from the subject;

(iii) optionally, means for establishing a subject profile of the quantity of MCAM and the presence or absence and/or quantity of said one or more other biomarkers; and

(iv) a reference profile of the quantity of MCAM and the presence or absence and/or quantity of said one or more other biomarkers, or means for establishing said reference profile, said reference profile representing a known diagnosis, prediction and/or prognosis of AHF.

30 . The kit according to any one of claims 26 to 29 , wherein the means for measuring the quantity of MCAM and/or the presence or absence and/or quantity of the one or more other biomarkers comprise, respectively, one or more binding agents capable of specifically binding to MCAM and/or to fragments thereof, and one or more binding agents capable of specifically binding to said one or more other biomarkers.

31 . The kit according to claim 30 , wherein the binding agent is an antibody, aptamer, photoaptamer, protein, peptide, peptidomimetic or a small molecule.

32 . The kit according to any one of claims 26 to 31 , configured as a portable device, preferably a bed-side device.

33 . The kit according to any one of claims 26 to 32 , wherein said means for detecting MCAM is capable of detecting the plasma-circulating form of MCAM, preferably capable of specifically detecting the plasma-circulating form of MCAM.

34 . A protein, polypeptide or peptide array or microarray comprising

(a) MCAM and/or a fragment thereof, preferably a known quantity or concentration of said MCAM and/or fragment thereof; and

(b) one or more other biomarkers useful for diagnosing, predicting and/or prognosticating AHF, preferably a known quantity or concentration of said one or more other biomarkers, and preferably wherein said other biomarkers are chosen from the group consisting of BNP, proBNP, NTproBNP and fragments of any one thereof.

35 . A binding agent array or microarray comprising:

(a) one or more binding agents capable of specifically binding to MCAM and/or to fragments thereof, preferably a known quantity or concentration of said binding agents; and

(b) one or more binding agents capable of specifically binding to one or more other biomarkers useful for diagnosing, predicting and/or prognosticating AHF, preferably a known quantity or concentration of said binding agents, and preferably wherein said other biomarkers are chosen from the group consisting of BNP, proBNP, NTproBNP and fragments of any one thereof.

36 . A portable testing device capable of measuring the quantity of MCAM in a sample from a subject comprising:

(i) means for obtaining a sample from the subject, and

(ii) means for measuring the quantity of MCAM in said sample, and

(iii) means for visualising the quantity of MCAM measured in the sample.

37 . The portable testing device according to claim 36 , wherein the visualising means is capable of indicating whether the quantity of MCAM in the sample is above or below a certain threshold level and/or whether the quantity of MCAM in the sample deviates or not from a reference value of the quantity of MCAM, said reference value representing a known diagnosis, prediction and/or prognosis of AHF.

38 . The portable testing device of claim 37 , further comprising said reference value or means for establishing said reference value.

39 . The portable device according to claim 37 , wherein the threshold level is chosen such that the quantity of MCAM in the sample above said threshold level indicates that the subject has or is at risk of having AHF or indicates a poor prognosis for AHF in the subject, and the quantity of MCAM in the sample below said threshold level indicates that the subject does not have or is not at risk of having AHF or indicates a good prognosis for AHF in the subject.

40 . The portable device according to claim 37 or 38 , comprising a reference value representing the diagnosis or prediction of no AHF or representing a good prognosis for AHF, or comprising means for establishing said reference value, such that an elevated quantity of MCAM in the sample from the subject compared to said reference value indicates that the subject has or is at risk of having AHF or indicates a poor prognosis for AHF in the subject.

41 . The portable device of any one of claims 36 to 40 , wherein said means of part (ii) and/or (iii) comprises a solid support having a proximal and distal end, comprising:

a sample application zone in the vicinity of the proximal end;

a reaction zone distal to the sample application zone; and

a detection zone distal to the reaction zone;

whereby said support has a capillary property that directs a flow of fluid sample applied in the application zone in a direction from the proximal end to the distal end.

42 . The portable device of claim 41 , wherein the reaction zone comprises one or more bands of a MCAM-specific binding molecules conjugated to a detection agent, which MCAM specific binding molecule conjugate is disposed on the solid support such that it can migrate with the capillary flow of fluid; and wherein the detection zone comprises one or more capture bands comprising a population of MCAM specific molecule immobilised on the solid support.

43 . The portable device of claim 42 , wherein the reaction zone additionally comprises one or more bands of capture MCAM-specific binding molecules in an amount sufficient to prevent a threshold quantity of MCAM specific binding molecule conjugates to migrate to the detection zone.

44 . An isolated fragment of MCAM.

45 . The isolated fragment of claim 44 , which is the plasma circulating fragment of MCAM.

46 . The isolated MCAM or fragment thereof according to claim 44 or 45 , comprising a detectable label.

47 . Use of isolated MCAM or fragment thereof according to any one of claims 44 - 46 , as a control, standard or calibator in a method according to any one of claims 1 to 25 .

48 . A testing device capable of measuring the quantity of MCAM in a sample from a subject comprising:

(i) means for measuring the quantity of MCAM in said sample, and

(ii) means of storing the reference value in the device, and

(iii) means of comparing the obtained quantity with the stored reference value, and

(iv) means for visualising the quantity of MCAM measured in the sample.