Lipidomic biomarkers for the prediction of cardiovascular outcomes in coronary artery disease patients undergoing statin treatment

Abstract

The present invention inter alia provides a method, and use thereof, of predicting severe CVD complications such as AMI or CVD death by detecting the lipid concentrations or lipid ratios of a biological sample and comparing it to a control and has identified specific lipid markers that are more specific and sensitive in predicting these CVD complications than currently utilized clinical markers. Also provided are antibodies towards said lipids, and the use thereof for predicting, diagnosing, preventing and/or treating CVD complications. The invention additionally relates to kits comprising lipids and/or an antibody thereto, for use in the prediction and/or diagnosis of CVD complications.

1 - 33 . (canceled) 
     
     
         34 . A method for determining whether a subject undergoing statin treatment is at risk to develop one or more CVD complications, comprising:
 a. determining in a sample from said subject one or more lipid-lipid concentration ratio(s), wherein (an) increased or decreased lipid-lipid concentration ratio(s) in said sample, when compared to a control sample, is (are) indicative of said subject having an increased risk of developing one or more CVD complications,   wherein the lipid-lipid concentration ratio whose increase is compared to the control is Cer(d18:1/20:0)/PC 18:1/20:4;   and wherein the one or more lipid-lipid concentration ratio(s) whose decrease(s) is (are) compared to the control is (are) selected from: CE 14:0/Cer(d18:1/20:0), CE 20:4/Cer(d18:1/20:0), CE 14:0/Cer(d18:1/16:0), CE 20:4/Cer(d18:1/16:0), CE 20:5/Cer(d18:1/20:0), CE 20:5/Cer(d18:1/24:1), CE 20:5/Cer(d18:1/16:0) and CE 20:5/Cer(d18:1/26:1); and/or   b. determining in a sample from said subject one or more lipid-clinical concentration ratio(s), wherein (a) decreased lipid-clinical concentration ratio(s) in said sample, when compared to a control sample, is (are) indicative of said subject having an increased risk of developing one or more CVD complications,   wherein the one or more lipid-clinical concentration ratio(s) whose decrease(s) is (are) compared to the control is (are) selected from: PC 16:0/18:1/supersensitive C-reactive protein, PC 18:0/20:3/supersensitive C-reactive protein, PC O-18:0/18:2-alkyl/supersensitive C-reactive protein, PC 16:0/16:0/supersensitive C-reactive protein, PC 18:0/18:2/supersensitive C-reactive protein, PC 18:1/20:4/supersensitive C-reactive protein, PC 18:1/18:2/supersensitive C-reactive protein, LPC 16:0/supersensitive C-reactive protein, PC-O 16:0/24-alkyl/CRP, PC O-16:0/18:2-alkyl/supersensitive C-reactive protein, PC 16:0/22:6/supersensitive C-reactive protein, PC 18:0/22:6/supersensitive C-reactive protein and Cer(d18:1/20:0)/total cholesterol).   
     
     
         35 . A method for determining whether a subject undergoing statin treatment who is not suffering from type 2 diabetes mellitus is at risk to develop one or more CVD complications, comprising:
 a. determining in a sample from said subject the concentration(s) of one or more lipid(s), wherein (an) increased or decreased concentration(s) in said sample, when compared to a control sample, is (are) indicative of said subject having an increased risk of developing one or more CVD complications,   wherein the lipid whose increase in concentration is compared to the control is PC 16:0/22:6;   and wherein the lipid whose decrease in concentration is compared to the control is PC 18:0/18:1;   b. determining in a sample from said subject one or more lipid-lipid concentration ratio(s), wherein (an) increased or decreased lipid-lipid concentration ratio(s) in said sample, when compared to a control sample, is (are) indicative of said subject having an increased risk of developing one or more CVD complications,   wherein the one or more lipid-lipid concentration ratio(s) whose increase(s) is compared to the control is (are) selected from: Cer(d18:1/26:1)/SM (d18:1/24:0) (d18:1/23:1-OH), PC 16:0/16:1/PC O-16:0/20:4-alkyl, PC 16:0/16:1/PC 18:2/18:2, PC 16:0/16:1/PC 18:1/20:4, PC 18:0/18:1/PC 18:2/18:2, Cer(d18:1/18:0)/PC O-16:0/20:4-alkyl, Cer(d18:1/20:0)/PC 18:2/18:2, Cer(d18:1/20:0)/PC O-16:0/20:4-alkyl, Cer(d18:1/24:1)/PC 18:2/18:2, Cer(d18:1/24:1)/PC 18:1/20:4, CE 16:1/PC 18:2/18:2, Cer(d18:1/18:0)/PC O-16:0/18:2-alkyl, PC 16:0/18:1/PC 18:1/20:4, PC 16:0/18:1/PC O-16:0/20:4-alkyl, CE 16:1/PC O-16:0/20:4-alkyl, PC 18:1/18:1/PC 18:1/20:4, Cer(d18:1/24:1)/PC O-16:0/18:2-alkyl, PC 18:0/18:1/PC 18:0/20:3, Cer(d18:1/26:1)/PC 16:0/22:6, CE 16:1/PC 18:1/20:4, Cer(d18:1/26:1)/PC 18:0/18:2, CE 16:1/Cer(d18:1/24:0), Cer(d18:1/26:1)/PC 18:1/18:2, Cer(d18:1/26:1)/PC 16:0/18:2, PC 18:1/18:2/PC 18:2/18:2 and PC 16:0/18:2/PC 18:2/18:2;   and wherein the one or more lipid-lipid concentration ratio(s) whose decrease(s) is (are) compared to the control is (are) selected from: CE 18:3/PC 16:0/18:1, CE 14:0/PC 18:0/18:1, CE 20:4/PC 18:0/18:1, Gb3(d18:1/22:0)/PC 16:0/18:1, CE 18:3/PC 18:0/18:1, CE 20:5/PC 16:0/16:1, CE 20:4/Cer(d18:1/26:1), CE 18:3/PC 16:0/16:1, CE 18:3/Cer(d18:1/26:1) and LPC 16:0/SM (d18:1/17:0) (d18:1/16:1-OH); and/or   c. determining in a sample from said subject one or more lipid-clinical concentration ratio(s), wherein (an) increased or decreased lipid-clinical concentration ratio(s) in said sample, when compared to a control sample, is (are) indicative of said subject having an increased risk of developing one or more CVD complications,   wherein the one or more lipid-clinical concentration ratio(s) whose increase(s) is (are) compared to the control is (are) selected from: PC 16:0/16:1/apolipoprotein A-I, PC 16:0/16:1/HDL cholesterol, PC 18:0/18:1/apolipoprotein A-I, PC 18:0/18:1/total cholesterol, PC 16:0/18:1/apolipoprotein A-I, PC 16:0/18:1/apolipoprotein A-I, PC 16:0/18:1/total cholesterol, PC 18:1/18:1/apolipoprotein A-I, PC 18:1/18:1/HDL cholesterol, PC 18:1/18:1/total cholesterol, PC 16:0/18:1/LDL cholesterol and PC 16:0/18:2/LDL cholesterol;   and wherein the one or more lipid-clinical concentration ratio(s) whose decrease(s) is (are) compared to the control is (are) selected from: PC 17:0/18:2/supersensitive C-reactive protein, PC 16:0/18:2/supersensitive C-reactive protein and PC 18:2/18:2/supersensitive C-reactive protein.   
     
     
         36 . A method for determining whether a subject undergoing statin treatment who is suffering from type 2 diabetes mellitus is at risk to develop one or more CVD complications, comprising:
 a. determining in a sample from said subject the concentration(s) of one or more lipid(s), wherein (an) increased or decreased concentration(s) in said sample, when compared to a control sample, is (are) indicative of said subject having an increased risk of developing one or more CVD complications,   wherein the one or more lipid(s) whose increase(s) in concentration is (are) compared to the control is (are) selected from: Cer(d18:1/16:0), Cer(d18:1/20:0) and PC 16:0/18:2;   and wherein the one or more lipid(s) whose decrease(s) in concentration is (are) compared to the control is (are) selected from: Cer(d18:1/24:0), PC 18:2/18:2, PC 18:0/22:6, PC P-18:0/20:4, PC O-16:0/20:4-alkyl, LPC 16:0, PC 18:1/20:4, PC 16:0/16:1, PC 16:0/16:0 and PC 18:1/18:2;   b. determining in a sample from said subject one or more lipid-lipid concentration ratio(s), wherein (an) increased or decreased lipid-lipid concentration ratio(s) in said sample, when compared to a control sample, is (are) indicative of said subject having an increased risk of developing one or more CVD complications,   wherein the one or more lipid-lipid concentration ratio(s) whose increase(s) is compared to the control is (are) selected from: Cer(d18:1/16:0)/DAG 16:0/18:1, Cer(d18:1/16:0)/PC 18:0/20:3, Cer(d18:1/20:0)/PC 18:0/20:3, PC 16:0/16:0/PC 18:1/20:4, PC 17:0/18:2/PC 18:1/20:4, PC 16:0/18:2/PC 18:1/20:4 and PC 18:0/18:2/PC 18:1/20:4;   and wherein the one or more lipid-lipid concentration ratio(s) whose decrease(s) is (are) compared to the control is (are) selected from: PC P-18:0/20:4/SM (d18:1/24:1) (d18:1/23:2-OH), CE 18:1/Cer(d18:1/16:0), CE 18:3/PC 18:0/18:2, PC 18:0/20:3/SM (d18:1/16:0) (d18:1/15:1-OH), PC 18:1/20:4/SM (d18:1/16:0) (d18:1/15:1-OH), PC 18:1/20:4/PC O-18:0/18:2-alkyl, CE 20:4/PC 17:0/18:2, PC 18:1/20:4/SM (d18:1/24:1) (d18:1/23:2-OH), PC 18:1/20:4/SM (d18:1/15:0) (d18:1/14:1-OH) and CE 16:1/Cer(d18:1/16:0); and/or   c. determining in a sample from said subject one or more lipid-clinical concentration ratio(s), wherein (an) increased or decreased lipid-clinical concentration ratio(s) in said sample, when compared to a control sample, is (are) indicative of said subject having an increased risk of developing one or more CVD complications,   wherein the one or more lipid-clinical concentration ratio(s) whose increase(s) is (are) compared to the control is (are) selected from: PC O-18:0/18:2-alkyl/apolipoprotein A-I and PC 17:0/18:2/triglycerides;   and wherein the one or more lipid-clinical concentration ratio(s) whose decrease(s) is (are) compared to the control is (are) selected from: PC P-18:0/20:4/apolipoprotein B, PC 18:1/20:4/HDL cholesterol, PC 16:0/16:1/apolipoprotein B, PC 18:1/20:4/total cholesterol, PC 18:0/20:3/apolipoprotein B, PC 18:1/20:4/apolipoprotein B, PC 18:0/20:3/LDL cholesterol, PC 18:1/20:4/LDL cholesterol, PC P-16:0/18:2/supersensitive C-reactive protein and PC P-18:0/20:4/supersensitive C-reactive protein.   
     
     
         37 . The method of  claim 34 , comprising determining at least 2, at least 3, at least 4, at least 5 or at least 6 lipid-lipid concentration ratio(s), lipid-clinical concentration ratio(s) or combinations thereof. 
     
     
         38 . The method of  claim 35 , comprising determining at least 2, at least 3, at least 4, at least 5 or at least 6 lipid concentration(s), lipid-lipid concentration ratio(s), lipid-clinical concentration ratio(s) or combinations thereof. 
     
     
         39 . The method of  claim 36 , comprising determining at least 2, at least 3, at least 4, at least 5 or at least 6 lipid concentration(s), lipid-lipid concentration ratio(s), lipid-clinical concentration ratio(s) or combinations thereof. 
     
     
         40 . The method of  claim 34 , wherein
 a. said CVD is characterized by coronary artery disease, peripheral artery disease, a stroke and/or CVD death; and/or   b. said CVD is atherosclerosis-induced; and/or   c. said subject has atherosclerosis; or   d. said subject does not have atherosclerosis.   
     
     
         41 . The method of  claim 35 , wherein
 a. said CVD is characterized by coronary artery disease, peripheral artery disease, a stroke and/or CVD death; and/or   b. said CVD is atherosclerosis-induced; and/or   c. said subject has atherosclerosis; or   d. said subject does not have atherosclerosis.   
     
     
         42 . The method of  claim 36 , wherein
 a. said CVD is characterized by coronary artery disease, peripheral artery disease, a stroke and/or CVD death; and/or   b. said CVD is atherosclerosis-induced; and/or   c. said subject has atherosclerosis; or   d. said subject does not have atherosclerosis.   
     
     
         43 . The method of  claim 34 , wherein
 a. the method further comprises determining the serum or plasma level of total cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), Apolipoprotein B (ApoB) and/or Apolipoprotein C-III (ApoC-III) in said sample; and/or   b. the subject does not have elevated serum or plasma levels of one or more of total cholesterol, low-density lipoprotein cholesterol (LDL-C), Apolipoprotein C-III (ApoC-III) or Apolipoprotein B (ApoB), or a decreased serum level of HDL-cholesterol (HDL-C).   
     
     
         44 . The method of  claim 35 , wherein
 a. the method further comprises determining the serum or plasma level of total cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), Apolipoprotein B (ApoB) and/or Apolipoprotein C-III (ApoC-III) in said sample; and/or   b. the subject does not have elevated serum or plasma levels of one or more of total cholesterol, low-density lipoprotein cholesterol (LDL-C), Apolipoprotein C-III (ApoC-III) or Apolipoprotein B (ApoB), or a decreased serum level of HDL-cholesterol (HDL-C).   
     
     
         45 . The method of  claim 36 , wherein
 a. the method further comprises determining the serum or plasma level of total cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), Apolipoprotein B (ApoB) and/or Apolipoprotein C-III (ApoC-III) in said sample; and/or   b. the subject does not have elevated serum or plasma levels of one or more of total cholesterol, low-density lipoprotein cholesterol (LDL-C), Apolipoprotein C-III (ApoC-III) or Apolipoprotein B (ApoB), or a decreased serum level of HDL-cholesterol (HDL-C).   
     
     
         46 . The method of  claim 34 , wherein
 a. the sample is blood, plasma, serum, urine or tissue, or a lipoprotein fraction thereof; and/or   b. the lipid concentration(s), the lipid-lipid concentration ratio(s) and/or the lipid-clinical concentration ratio(s) is (are) determined by using mass spectrometry, nuclear magnetic resonance spectroscopy, fluorescence spectroscopy or dual polarisation interferometry, a high performance separation method, an immunoassay and/or with a binding moiety capable of specifically binding the analyte.   
     
     
         47 . The method of  claim 35 , wherein
 a. the sample is blood, plasma, serum, urine or tissue, or a lipoprotein fraction thereof; and/or   b. the lipid concentration(s), the lipid-lipid concentration ratio(s) and/or the lipid-clinical concentration ratio(s) is (are) determined by using mass spectrometry, nuclear magnetic resonance spectroscopy, fluorescence spectroscopy or dual polarisation interferometry, a high performance separation method, an immunoassay and/or with a binding moiety capable of specifically binding the analyte.   
     
     
         48 . The method of  claim 36 , wherein
 a. the sample is blood, plasma, serum, urine or tissue, or a lipoprotein fraction thereof; and/or   b. the lipid concentration(s), the lipid-lipid concentration ratio(s) and/or the lipid-clinical concentration ratio(s) is (are) determined by using mass spectrometry, nuclear magnetic resonance spectroscopy, fluorescence spectroscopy or dual polarisation interferometry, a high performance separation method, an immunoassay and/or with a binding moiety capable of specifically binding the analyte.   
     
     
         49 . The method of  claim 34 , wherein the one or more CVD complications including are selected from CVD death and acute myocardial infarction (AMI). 
     
     
         50 . The method of  claim 35 , wherein the one or more CVD complications including are selected from CVD death and acute myocardial infarction (AMI). 
     
     
         51 . The method of  claim 36 , wherein the one or more CVD complications including are selected from CVD death and acute myocardial infarction (AMI). 
     
     
         52 . The method of  claim 34 , wherein the subject is at risk to develop or has suffered from one or more CVD complications. 
     
     
         53 . The method of  claim 35 , wherein the subject is at risk to develop or has suffered from one or more CVD complications. 
     
     
         54 . The method of  claim 36 , wherein the subject is at risk to develop or has suffered from one or more CVD complications. 
     
     
         55 . The method of  claim 34 , wherein the control sample is from (a) CAD patient(s) or a group of CAD patients, wherein the CAD patient(s) or group of CAD patients has/have no history of major CVD events and is/are undergoing statin treatment; wherein the control sample is blood, plasma, serum, urine or tissue, or a lipoprotein fraction thereof. 
     
     
         56 . The method of  claim 35 , wherein the control sample is from (a) CAD patient(s) or a group of CAD patients, wherein the CAD patient(s) or group of CAD patients has/have no history of major CVD events and is/are undergoing statin treatment; wherein the control sample is blood, plasma, serum, urine or tissue, or a lipoprotein fraction thereof. 
     
     
         57 . The method of  claim 36 , wherein the control sample is from (a) CAD patient(s) or a group of CAD patients, wherein the CAD patient(s) or group of CAD patients has/have no history of major CVD events and is/are undergoing statin treatment; wherein the control sample is blood, plasma, serum, urine or tissue, or a lipoprotein fraction thereof. 
     
     
         58 . The method of  claim 34 , further comprising:
 determining that the subject is at risk to develop one or more CVD complications; and treating the subject.   
     
     
         59 . The method of  claim 35 , further comprising:
 determining that the subject is at risk to develop one or more CVD complications; and treating the subject.   
     
     
         60 . The method of  claim 36 , further comprising:
 determining that the subject is at risk to develop one or more CVD complications; and treating the subject.   
     
     
         61 . A method of treating or preventing CVD complications in a subject undergoing statin treatment identified as being at risk to develop one or more CVD complications, the method comprising:
 administering a treatment to the subject, wherein, prior administering the treatment, the subject has been identified as being at risk to develop one or more CVD complications by the method of  claim 34 .   
     
     
         62 . A method of treating or preventing CVD complications in a subject undergoing statin treatment identified as being at risk to develop one or more CVD complications, the method comprising:
 administering a treatment to the subject, wherein, prior administering the treatment, the subject has been identified as being at risk to develop one or more CVD complications by the method of  claim 35 .   
     
     
         63 . A method of treating or preventing CVD complications in a subject undergoing statin treatment identified as being at risk to develop one or more CVD complications, the method comprising:
 administering a treatment to the subject, wherein, prior administering the treatment, the subject has been identified as being at risk to develop one or more CVD complications by the method of  claim 36 .   
     
     
         64 . The method of  claim 61 , wherein the treatment is a change in, or supplementation of the already administered statin treatment. 
     
     
         65 . The method of  claim 62 , wherein the treatment is a change in, or supplementation of the already administered statin treatment. 
     
     
         66 . The method of  claim 63 , wherein the treatment is a change in, or supplementation of the already administered statin treatment.