Blood based biomarkers for diagnosing atherosclerotic coronary artery disease

Abstract

The invention, in some aspects, relates to methods for evaluating a human subject for having atherosclerotic coronary artery disease (ASCAD) or as having a coronary atherosclerotic plaque. In some aspects, the invention relates to methods and kits useful for diagnosing, classifying, profiling and treating atherosclerotic CAD and or a coronary atherosclerotic plaque.

1 . A method for evaluating a human subject for having atherosclerotic coronary artery disease (ASCAD) or as having a coronary atherosclerotic plaque, the method comprising:
 measuring the levels of each analyte within a panel of analyte biomarkers in a biological sample obtained from a subject, wherein the panel of analyte biomarkers is selected from the group consisting of:   (i) a panel of biomarkers comprising valylleucine, glutamate, acisoga, urate, glucuronate, fucose, butyrylcarnitine (C4), mannose;   (ii) a panel of biomarkers comprising acisoga, o.cresol.sulfate, threonate, cysteine.glutathione.disulfide;   (iii) a panel of biomarkers comprising N.acetylphenylalanine, N.acetylleucine, valylleucine, xylitol, 2.hydroxyhippurate (salicylurate), glycylphenylalanine, serylleucine, fucose;   (iv) a panel of biomarkers comprising glutamate, acisoga, valylleucine, mannose, glucuronate, urate, valerylcarnitine (C5), 12.HETE;   (v) a panel of biomarkers comprising o.cresol.sulfate, acisoga, cysteine.glutathione.disulfide, glucose, 1.nonadecanoylglycerophosphocholine (19:0), threonate;   (vi) a panel of biomarkers comprising N.acetylphenylalanine, N.acetylleucine, glycylphenylalanine, valylleucine, xylitol, 12.HETE, 2.hydroxyhippurate (salicylurate);   (vii) a panel of biomarkers comprising N.acetylthreonine, tigloylglycine, glycerate, valerylcarnitine (C5), threonate, 2.hydroxyhippurate (salicylurate), salicylate, tartronate (hydroxymalonated);   (viii) a panel of biomarkers comprising N.acetylglycine, 3.ethylphenylsulfate;   (ix) a panel of biomarkers comprising serotonin (5HT), N.acetylputrescine, leucylleucine, propionylglycine (C3), cholate, glycocholenate.sulfate, asparagine, 3.ethylphenylsulfate;   (x) a panel of biomarkers comprising acisoga, mannose, valerylcarnitine (C5), 1.linoleoyl.GPE (18.2), glycoursodeoxycholate, threonate, 2.hydroxyhippurate (salicylurate), salicylate;   (xi) a panel of biomarkers comprising N.acetylglycine, threonate, 2.hydroxyhippurate (salicylurate), 3.ethylphenylsulfate, salicylate;   (xii) a panel of biomarkers comprising asparagine, taurine, acisoga, 1.oleoylglycerol (18:1), cholesterol, 2.hydroxyhippurate (salicylurate), 6.oxopiperidine.2.carboxylic.acid, propionylglycine (C3);   comparing the measured levels of the analytes in the biological sample with one or more reference samples, wherein the references are representative of matched human subjects; and   identifying the subject as having atherosclerotic coronary artery disease or as having a coronary atherosclerotic plaque if measured levels of the analytes in the biological sample are increased or decreased relative to the amounts of the analytes in the reference samples.   
     
     
         2 . The method of  claim 1 , further comprising instructing a healthcare professional to complete a non-invasive cardiovascular evaluation on a subject identified as having atherosclerotic coronary artery disease or as having a coronary atherosclerotic plaque to confirm the subject has or does not have atherosclerotic coronary artery disease or a coronary atherosclerotic plaque. 
     
     
         3 . The method of  claim 1 , further comprising performing a non-invasive cardiovascular evaluation on a subject identified as having atherosclerotic coronary artery disease or as having a coronary atherosclerotic plaque to confirm the subject does or does not have atherosclerotic coronary artery disease or a coronary atherosclerotic plaque. 
     
     
         4 . The method of  claim 2 , wherein performing a non-invasive cardiovascular evaluation comprises performing a procedure selected from the group consisting of cardiovascular computed tomography (CT) imaging, an exercise stress test, a pharmacologic stress test, myocardial perfusion imaging, stress echocardiography, and cardiovascular magnetic resonance imaging. 
     
     
         5 . The method of  claim 1 , further comprising selectively administering a composition comprising an effective amount of a therapeutic agent selected from the group consisting of a statin, cholesterol absorption inhibitors, niacin-derivatives, omega-3-fatty acid compounds, bile acid sequestrants, anti-platelet agents, aldosterone blockers, angiotensin-converting enzyme (ACE) inhibitors, angiotensin-receptor blockers (ARBs), beta blockers, diuretics, digitalis, hydralazine and nitrates, warfarin and aspirin to a subject identified as having atherosclerotic coronary artery disease or as having a coronary atherosclerotic plaque to treat the subject. 
     
     
         6 . The method of  claim 1 , further comprising selecting a treatment plan for a subject identified as having atherosclerotic coronary artery disease or as having a coronary atherosclerotic plaque to treat the subject. 
     
     
         7 . The method of  claim 6 , wherein the treatment plan comprises selectively administering a composition comprising administering an effective amount of a therapeutic agent selected from the group consisting of a statin, cholesterol absorption inhibitors, niacin-derivatives, omega-3-fatty acid compounds, bile acid sequestrants, anti-platelet agents, aldosterone blockers, angiotensin-converting enzyme (ACE) inhibitors, angiotensin-receptor blockers (ARBs), beta blockers, diuretics, digitalis, hydralazine and nitrates, warfarin and aspirin. 
     
     
         8 . The method of  claim 1 , wherein the levels are measured using mass spectrometry (MS) analysis. 
     
     
         9 . The method of  claim 8 , wherein the MS analysis comprises liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) or gas chromatography-mass spectrometry (GC-MS). 
     
     
         10 . The method of  claim 1 , wherein the CAD comprises atherosclerosis. 
     
     
         11 . The method of  claim 1 , wherein the biological sample is a blood sample, a serum sample or a plasma sample. 
     
     
         12 . The method of  claim 1 , wherein the subject presents with symptoms of atherosclerotic CAD comprising chest pain, angina, angina equivalent, dyspnea, or dyspnea on exertion. 
     
     
         13 . The method of  claim 1 , wherein the subject presents with risk factors associated with coronary artery disease selected from the group consisting of male gender, hypertension, dyslipidemia, diabetes, and a family history of coronary artery disease. 
     
     
         14 . The method of  claim 1 , wherein the subject has no previous history of coronary artery disease. 
     
     
         15 . The method of  claim 1 , wherein the reference sample is obtained from at least one individual not suffering from a cardiovascular disease. 
     
     
         16 . The method of  claim 1 , wherein the reference sample comprises predetermined, statistically significant reference analyte levels. 
     
     
         17 . The method of  claim 1 , wherein the determining step is performed using one of the multivariate methods, such as a gradient boosting algorithm, decision tree models, or by other generalized linear regression analyses. 
     
     
         18 . The method of  claim 1 , further comprising modifying the subject's clinical record to identify the subject as having atherosclerotic coronary artery disease or as having a coronary atherosclerotic plaque. 
     
     
         19 . The method of  claim 18 , wherein the clinical record is stored in a computer readable medium. 
     
     
         20 . The method of  claim 1 , further comprising selecting a subject identified as having atherosclerotic coronary artery disease or as having a coronary atherosclerotic plaque for treatment, wherein the treatment comprises selectively administering a composition comprising an effective amount of a therapeutic agent selected from the group consisting of a statin, cholesterol absorption inhibitors, niacin-derivatives, omega-3-fatty acid compounds, bile acid sequestrants, anti-platelet agents, aldosterone blockers, angiotensin-converting enzyme (ACE) inhibitors, angiotensin-receptor blockers (ARBs), beta blockers, diuretics, digitalis, hydralazine and nitrates, warfarin and aspirin to a subject identified as having atherosclerotic coronary artery disease or as having a coronary atherosclerotic plaque to treat the subject. 
     
     
         21 . The method of  claim 1 , further comprising selecting a subject identified as having atherosclerotic coronary artery disease or as having a coronary atherosclerotic plaque for treatment, wherein the treatment comprises performing a non-invasive cardiovascular evaluation on a subject identified as having atherosclerotic coronary artery disease or as having a coronary atherosclerotic plaque to confirm the subject does or does not have atherosclerotic coronary artery disease or a coronary atherosclerotic plaque. 
     
     
         22 . The method of  claim 6 , wherein the treatment plan comprises prescribing therapeutic lifestyle changes. 
     
     
         23 . The method of  claim 6 , wherein the treatment plan comprises performing a non-invasive cardiovascular evaluation comprises performing a procedure selected from the group consisting of cardiovascular computed tomography (CT) imaging, an exercise stress test, a pharmacologic stress test, myocardial perfusion imaging, stress echocardiography, and cardiovascular magnetic resonance imaging. 
     
     
         24 . The method of  claim 1 , wherein the comparing step includes analysis of the subject's age and/or gender. 
     
     
         25 . A kit for evaluating a human subject for having atherosclerotic coronary artery disease (ASCAD) or as having a coronary atherosclerotic plaque, the kit comprising:
 reagents suitable for determining levels of a plurality of analytes in a test sample, wherein the plurality of analytes comprise:   (i) a panel of biomarkers comprising valylleucine, glutamate, acisoga, urate, glucuronate, fucose, butyrylcarnitine (C4) and mannose;   (ii) a panel of biomarkers comprising acisoga, o.cresol.sulfate, threonate, cysteine.glutathione.disulfide;   (iii) a panel of biomarkers comprising N.acetylphenylalanine, N.acetylleucine, valylleucine, xylitol, 2.hydroxyhippurate (salicylurate), glycylphenylalanine, serylleucine and fucose;   (iv) a panel of biomarkers comprising glutamate, acisoga, valylleucine, mannose, glucuronate, urate, valerylcarnitine (C5) and 12.HETE;   (v) a panel of biomarkers comprising o.cresol.sulfate, acisoga, cysteine.glutathione.disulfide, glucose, 1.nonadecanoylglycerophosphocholine (19:0) and threonate;   (vi) a panel of biomarkers comprising N.acetylphenylalanine, N.acetylleucine, glycylphenylalanine, valylleucine, xylitol, 12.HETE and 2.hydroxyhippurate (salicylurate);   (vii) a panel of biomarkers comprising N.acetylthreonine, tigloylglycine, glycerate, valerylcarnitine (C5), threonate, 2.hydroxyhippurate (salicylurate), salicylate and tartronate (hydroxymalonated);   (viii) a panel of biomarkers comprising N.acetylglycine and 3.ethylphenylsulfate;   (ix) a panel of biomarkers comprising serotonin (5HT), N.acetylputrescine, leucylleucine, propionylglycine (C3), cholate, glycocholenate.sulfate, asparagine and 3.ethylphenylsulfate;   (x) a panel of biomarkers comprising acisoga, mannose, valerylcarnitine (C5), 1.linoleoyl.GPE (18.2), glycoursodeoxycholate, threonate, 2.hydroxyhippurate (salicylurate) and salicylate;   (xi) a panel of biomarkers comprising N.acetylglycine, threonate, 2.hydroxyhippurate (salicylurate), 3.ethylphenylsulfate and salicylate;   (xii) a panel of biomarkers comprising asparagine, taurine, acisoga, 1.oleoylglycerol (18:1), cholesterol, 2.hydroxyhippurate (salicylurate), 6.oxopiperidine.2.carboxylic.acid and propionylglycine (C3);   optionally one or more control samples comprising predetermined levels of the same analytes, wherein comparison of the levels of the analytes in a test sample with levels in the control samples identifies a subject as having atherosclerotic coronary artery disease or as having a coronary atherosclerotic plaque; and   instructions for use of the kit in the method according to  claim 1 .