Biomarkers for cardiovascular events

Abstract

To provide methods and uses relating to determination of lipid concentration for diagnosing, predicting, preventing and / or treating one or more cardiovascular events in a subject.SOLUTION: A method comprises analyzing lipid concentrations of a sample from a subject and comparing them to a control.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to the field of prognostic biomarkers for cardiovascular (CV) outcome.In particular, the present disclosure provides a novel in vitro method for evaluating the risk of subject developing cardiovascular events.In addition, the biomarkers of the present invention can be used in the method for evaluating the effectiveness of treatment of cardiovascular disease (CVD), treating cardiovascular disease, and preventing cardiovascular events. [Background technology]

[0002] Cardiovascular disease is one of the leading causes of mortality and morbidity worldwide and continues to become ever more prevalent. CVD is used to classify many conditions that affect the heart, heart valves, blood, and vascular system of the body. For a decade, plasma or serum total cholesterol, low-density lipoprotein (LDL) or high-density lipoprotein (HDL) concentrations have been used as gold standard biomarkers for predicting CVD risk and targeting treatment. However, recent studies have shown that these widely used biomarkers are not reliably associated with CV outcomes, such as myocardial infarction (MI) or cardiovascular death. For example, it has been observed that half of patients with acute myocardial infarction (AMI) have LDL-cholesterol (LDL-C) levels within the recommended normal range. Nevertheless, reduction of LDL-C concentrations is currently used as the primary target of CVD treatment, such as the widely used statin treatment. In addition, it has been recognized that despite the reduction of LDL-C, there remains a substantial risk of developing CVD events in patients treated with statins. Thus, there is a need for additional prognostic and therapeutic targets beyond LDL-C. In addition to total cholesterol, LDL-C, and HDL-C, several non-lipid risk factors (including age, blood pressure, diabetes, smoking, and body mass index) are used in risk assessments to assess an individual's risk for cardiovascular events. Statins are a family of lipid-lowering drugs for people at high risk of cardiovascular events. Statins are widely used. For example, there are nearly 20 million patients treated with statins in the United States alone. In addition, it is calculated that nearly 50 million patients in the United States alone would benefit from statin treatment. However, despite statin treatment, CVD patients have a substantial risk of developing severe CVD-related events. Early initiation of preventive measures targeting CVD-related severe events, such as AMI and cardiovascular death, would have significant benefits and provide a major opportunity to reduce mortality and morbidity in patients suffering from CVD. Accurate identification of individuals at risk of developing CVD and cardiovascular events is necessary. Conventional risk assessment classifies the majority of individuals as having moderate risk, but fails to recognize the majority of high-risk patients, leaving the management of patients uncertain. Therefore, additional strategies to further improve risk assessment of CVD patients are greatly needed. For example, Zora Biosciences patent applications WO2011138419, WO2011161062, WO2013068373 and WO2013068374 have identified a large group of lipid molecules for predicting cardiovascular events in certain subject populations. However, there remains a need for improved methods for identifying individuals at risk of developing cardiovascular events. Summary of the Invention

[0003] The biomarker combination of the present disclosure provides superior performance for risk stratification compared to any other lipid-based cardiovascular biomarkers currently in use. In addition, the levels of the biomarker combination of the present disclosure may be affected by certain lipid-modifying therapies, such as statins. Thus, the biomarker combination of the present disclosure provides accurate and actionable risk stratification of cardiovascular events.

[0004] The present disclosure provides novel biomarker combinations and related diagnostic methods and uses for identifying subjects at risk of developing cardiovascular events. Such methods and uses include monitoring combinations of specific lipid concentrations in samples from subjects and comparing such concentrations with those of controls. The novel biomarker combination consists of at least one ceramide (Cer) molecule and at least one phosphatidylcholine (PC) molecule, wherein the at least one ceramide is selected from the ceramides of formula I and the at least one phosphatidylcholine is selected from the phosphatidylcholine of formula II.

[0005] Ceramides of formula I have the following structure: [ka] (In the formula, R 1 is a saturated, monounsaturated or diunsaturated alkyl chain having 11 to 17 carbon atoms, R 2 is a saturated, monounsaturated or diunsaturated alkyl chain having 13 to 25 carbon atoms.

[0006] Phosphatidylcholine of formula II has the following structure: [ka] (In the formula, R 3 and R 4 is a saturated, monounsaturated or polyunsaturated alkyl chain)

[0007] In a first aspect of the present disclosure, there is provided a method for determining a subject's risk of developing one or more cardiovascular events, comprising assaying a sample from said subject to determine a concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject is at risk of developing one or more cardiovascular events. In certain embodiments of the method for determining a subject's risk of developing one or more cardiovascular events, at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II.

[0008] In certain embodiments, the method further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, where an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to a control indicates that the subject is at risk for developing one or more cardiovascular events.

[0009] Table 1. Additional ceramides (Cer, Table 1a) and phosphatidylcholines (PC, Table 1b). According to all embodiments of the present disclosure, PC can be brutto PC or molecular PC. According to the present disclosure, molecular PC is not limited to the examples presented in Table 1. The direction of change refers to the change in the amount, content, abundance, level or concentration of the biomarker in a sample from a subject compared to a control when the subject is at risk of developing one or more cardiovascular events. When evaluating the effectiveness of a CVD treatment in a subject, according to some embodiments of the present disclosure, the direction of change is the opposite direction to that presented in Table 1. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0010] In certain embodiments, the method of determining a subject's risk of developing one or more cardiovascular events further comprises, after the determining step and / or after the step of assaying at least one additional Cer and / or at least one additional PC, a step of diagnosing the subject, e.g., a human subject, as having a risk of developing one or more cardiovascular events, and a step of administering a treatment to the subject diagnosed in the previous step. In certain embodiments, the method for determining a subject's risk of developing one or more cardiovascular events further comprises, after the determining step and / or after the assaying step of at least one additional Cer and / or at least one additional PC, a step of diagnosing the subject, e.g., a human subject, as having a risk of developing one or more cardiovascular events, and a step of administering a drug, and / or a step of providing a treatment, behavioral and / or lifestyle modification to the subject diagnosed in the previous step.

[0011] In one aspect, the present disclosure is directed to a method of treating cardiovascular disease (CVD) or preventing one or more cardiovascular (CV) events in a subject identified as having a risk of developing one or more CV events, comprising administering to the subject a treatment described herein, where prior to administering the treatment, the subject is identified as having a risk of developing one or more CV events by a method of determining risk described herein.

[0012] In another aspect, the present disclosure is directed to a method of treating CVD or preventing one or more CV events in a subject, comprising determining a subject's risk of developing one or more CV events according to the methods for determining risk described herein, and administering treatment to the subject if the subject is identified as at risk of developing one or more CV events.

[0013] In accordance with the present disclosure, treatment may include, for example, administering a drug to the subject and / or providing a therapeutic, behavioral and / or lifestyle modification. The drug may be, for example, a statin, another lipid-lowering drug, and / or a lipid concentration modulator as described elsewhere in this disclosure. Behavioral and / or lifestyle modification may include, for example, lifestyle counseling, including, but not limited to, instructions and / or encouragement regarding a healthy diet, physical activity / exercise, and / or smoking cessation. Treatment may also be a surgical procedure as described herein. In certain embodiments, a method of treating CVD or preventing one or more CV events in a subject further includes identifying a subject as in need of treatment or prevention, e.g., by requesting a test or receiving test results from a commercial testing laboratory that provides assay results useful for determining the concentration of at least one Cer of formula I and at least one PC of formula II, and administering a treatment to the subject, e.g., administering a therapeutically effective amount of a drug, if the subject has an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control.

[0014] In another aspect of the present disclosure, a method is provided for detecting at least two lipids in a sample from a subject, the method comprising detecting the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II.

[0015] In certain embodiments of the method comprising detecting as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II. In certain embodiments, the method further comprises detecting the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1 in the sample. In certain embodiments, the method further comprises comparing the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II, and / or at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1, with a control.

[0016] In certain embodiments, the method includes detecting an altered concentration of at least one Cer of formula I and / or an altered concentration of at least one PC of formula II in a sample from a subject, e.g., compared to a control. In certain embodiments, the method includes detecting an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in a sample from a subject, e.g., compared to a control. In certain embodiments, the method for detecting the concentration of at least one Cer of formula I and at least one PC of formula II in a sample from a subject comprises using a standard combination comprising at least one Cer of formula I and / or at least one PC of formula II. In certain embodiments, the standard combination comprises at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II. In certain embodiments, the isotopes of at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II are deuterium, 13 C or 15 It's N. In certain embodiments, the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1. In certain embodiments, at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 is isotopically labeled. In certain embodiments, the isotope of at least one additional isotopically labeled Cer and / or at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1 is deuterium, 13 C or 15 It's N. In certain embodiments, the method further comprises comparing the concentration of at least one Cer of formula I and the concentration of at least one PC of formula II with a control, and determining that the subject is at risk of developing one or more CV events if the concentration of at least one Cer of formula I is increased and / or the concentration of at least one PC of formula II is decreased compared to the control.

[0017] In another aspect of the present disclosure, there is provided a method for obtaining data for determining a subject's risk of developing one or more CV events, comprising assaying a sample from a subject to determine a concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject is at risk of developing one or more cardiovascular events. In certain embodiments of the methods of obtaining data as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II. In certain embodiments, the method further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, where an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to a control indicates that the subject is at risk for developing one or more cardiovascular events.

[0018] In another aspect of the disclosure, a method is provided for generating quantitative data regarding a subject, the method comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II. In certain embodiments of the methods of generating quantitative data as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the method further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1.

[0019] In another aspect of the present disclosure, a method is provided for assessing the effectiveness of a CVD treatment in a subject, comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein a decreased concentration of at least one Cer of formula I and / or an increased concentration of at least one PC of formula II in the sample compared to a control indicates the effectiveness of said treatment.

[0020] In certain embodiments of the methods of assessing the effectiveness of a CVD treatment as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the method further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC compared to a control indicates the effectiveness of said treatment. In certain embodiments, the method for assessing the effectiveness of a CVD treatment in a subject according to the present disclosure may further comprise, after the determining step and / or after the assaying step of at least one additional Cer and / or at least one additional PC biomarker, modifying, supplementing, or maintaining a treatment already administered to the subject based on the Cer concentration and / or PC concentration obtained in the determining step and / or the assaying step of at least one additional Cer and / or at least one additional PC biomarker. In certain embodiments, the method of assessing the effectiveness of a CVD treatment in a subject further comprises determining that the treatment is not effective in the subject and increasing or changing the subject's treatment to a different treatment. In certain embodiments, escalation of the treatment regimen is based on unfavorable changes in the concentration of at least one Cer of formula I and / or at least one PC of formula II obtained in the determining step, as defined in the relevant guidelines, to move the subject to the next recommended step. In certain embodiments, the treatment regimen determined to be ineffective comprises administration of a drug, e.g., a lipid-lowering drug (e.g., a statin) as described herein. In certain embodiments, the associated guidelines comprise increasing the dose or type of drug administered to the subject compared to the dose or type of drug administered prior to the determining step.

[0021] In another aspect, the present disclosure is directed to a method of treating CVD or preventing one or more CV events in a subject undergoing treatment for CVD, e.g., coronary artery disease or atherosclerosis, comprising administering to the subject a drug, e.g., a lipid-lowering drug as described herein, and / or providing a therapeutic, behavioral and / or lifestyle modification, wherein prior to administration of the drug, the subject is identified as being treated ineffectively for CVD by a method for assessing the effectiveness of treatment as described herein.

[0022] In another aspect, the present disclosure is directed to a method of treating CVD or preventing one or more CV events in a subject receiving treatment for CVD, e.g., coronary artery disease or atherosclerosis, comprising determining whether the subject is being treated ineffectively for CVD according to a method for assessing the effectiveness of treatment as described herein, and if the subject is identified as being treated ineffectively for CVD, administering a drug, e.g., a lipid-lowering drug, and / or providing a therapeutic, behavioral and / or lifestyle modification to the subject.

[0023] In another embodiment of the present disclosure, a method is provided for selecting an appropriate CVD treatment for a subject, comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject requires treatment, or a modification or supplementation of an already administered treatment.

[0024] In certain embodiments of the method of selecting an appropriate CVD treatment for a subject as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the method further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, where an increased or decreased concentration of at least one additional Cer and / or at least one additional PC as set forth in Table 1 compared to a control indicates that the subject requires treatment, or a change or supplementation of an already administered treatment. In certain embodiments, the methods provided for selecting an appropriate CVD treatment for a subject further comprise, after the determining step and / or after the step of assaying at least one additional Cer and / or at least one additional PC biomarker, treating the subject and / or providing the subject with a therapeutic, behavioral and / or lifestyle modification based on the concentration of Cer and / or the concentration of PC obtained in the determining step. In certain embodiments, the method of selecting an appropriate CVD treatment for a subject further comprises, after the determining step and / or after the assaying step of at least one additional Cer and / or at least one additional PC biomarker, determining that the subject requires treatment, or a modification or supplementation of an already administered treatment, and administering to the subject the treatment determined to be required by the subject, and / or providing the subject with a therapeutic, behavioral and / or lifestyle modification. In certain embodiments, the CVD treatment whose effectiveness may be evaluated or optionally selected according to the methods described and claimed herein is a lipid-modifying therapy (e.g., a statin or other lipid-lowering drug described elsewhere in this application).

[0025] In another aspect, the present disclosure is directed to a method of treating CVD or preventing one or more CV events in a subject, comprising administering to the subject a drug, e.g., a lipid-lowering drug as described herein, and / or providing a therapeutic, behavioral and / or lifestyle modification, wherein prior to administering the drug and / or providing the therapeutic, behavioral and / or lifestyle modification, the subject is identified as needing a modification of, or supplementation of, an already administered treatment by a method of selecting an appropriate CVD treatment as described herein.

[0026] In another aspect, the present disclosure is directed to a method of treating CVD or preventing one or more CV events in a subject, comprising determining whether the subject requires modification or supplementation of a treatment already being administered according to a method for selecting an appropriate treatment as described herein, and administering to the subject a drug, e.g., a lipid-lowering drug as described herein, if the subject is identified as requiring modification or supplementation of a treatment already being administered.

[0027] Another aspect of the present disclosure relates to a method of treating CVD or preventing one or more CV events in a subject, comprising administering to the subject a therapeutically effective amount of a drug, wherein the drug is a lipid-lowering drug selected from a statin; an HMG-CoA reductase inhibitor other than a statin, niacin (nicotinic acid), a cholesterol absorption inhibitor (e.g., ezetimibe), a cholesteryl ester transfer protein (CETP), a bile acid sequestrant, a fibrate, a phytosterol, an anti-inflammatory agent (e.g., methotrexate, IL-1 mAb, TNF-alpha mAb), acetylsalicylic acid (aspirin), and a PCSK9 inhibitor; or a modulator of lipid levels selected from a small molecule, an antibody, an antisense RNA, a small interfering RNA (siRNA), and a natural or modified lipid, and wherein prior to administering the drug, the subject is identified as being at risk for developing one or more CV events based on the methods disclosed herein. According to any of the aspects and embodiments of the present disclosure, the subject can be or has been treated with a statin, another lipid-lowering drug, and / or a modulator of lipid concentrations. Alternatively, the subject may be naive to statin therapy, treatment with another lipid-lowering drug, and / or treatment with a modulator of lipid concentrations.

[0028] In some embodiments, the treatment or prevention method further includes identifying a subject as in need of treatment or prevention by, for example, requesting testing by a commercial laboratory, providing the results of an assay useful for determining the concentration of at least one Cer of formula I and / or at least one PC of formula II, and administering a treatment to the subject, for example, administering a therapeutically effective amount of a drug and / or administering a therapeutic, behavioral and / or lifestyle modification, if the subject has an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II compared to a control sample.

[0029] In yet another aspect of the present disclosure, there is provided a method for collecting data for determining a subject's risk of developing one or more cardiovascular events, comprising assaying a sample from the subject to determine a concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

[0030] In certain embodiments of the methods of collecting data as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 phosphatidylcholines of formula II. In certain embodiments, the method further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, where an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to a control indicates that the subject is at risk for developing one or more cardiovascular events.

[0031] In another aspect of the present disclosure, a method is provided for collecting data to detect the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample from a subject. In certain embodiments of the method of collecting data for detection as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the method further comprises detecting the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1 in the sample.

[0032] In another aspect of the present disclosure, a method is provided for collecting data for assessing the effectiveness of a CVD treatment in a subject, comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein a decreased concentration of at least one Cer of formula I and / or an increased concentration of at least one PC of formula II in the sample compared to a control indicates the effectiveness of said treatment.

[0033] In certain embodiments of the methods of collecting data for evaluating the effectiveness of a CVD treatment as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 ceramides of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the method further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC compared to a control indicates the effectiveness of said treatment.

[0034] In another embodiment of the present disclosure, a method is provided for collecting data to select an appropriate CVD treatment for a subject, comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject requires treatment, or a modification or supplementation of an already administered treatment.

[0035] In certain embodiments of the method of collecting data to select an appropriate CVD treatment as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the method further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, where an increased or decreased concentration of at least one additional Cer and / or at least one additional PC as set forth in Table 1 compared to a control indicates that the subject requires treatment, or a change or supplementation of an already administered treatment.

[0036] In another aspect of the present disclosure, there is provided a cardiovascular disease (CVD) marker combination for determining a subject's risk of developing one or more cardiovascular events, the CVD marker combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II. In certain embodiments of the CVD marker combinations as described herein, at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the CVD marker combination further comprises at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1.

[0037] In another aspect of the present disclosure, there is provided a use of a CVD marker combination for determining a subject's risk of developing one or more cardiovascular events, the CVD marker combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising assaying a sample from the subject to determine a concentration of at least one Cer of formula I and at least one PC of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject is at risk of developing one or more cardiovascular events. In certain embodiments of the use of the CVD marker combination to determine a subject's risk of developing one or more cardiovascular events as described herein, at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II, and the CVD marker combination comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use of the CVD marker combination further comprises assaying the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, where an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to a control indicates that the subject is at risk of developing one or more cardiovascular events, and the CVD marker combination further comprises at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1.

[0038] In another aspect of the present disclosure, there is provided a use of a CVD marker combination for detecting the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample from a subject.

[0039] In certain embodiments of the use of a CVD marker combination for detection as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use further comprises detecting the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1 in the sample. In certain embodiments, the use includes detecting an altered concentration of at least one Cer of formula I and / or an altered concentration of at least one PC of formula II in a sample from a subject, e.g., compared to a control. In certain embodiments, the use comprises detecting an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in a sample from a subject.

[0040] In another aspect of the present disclosure, there is provided a use of a CVD marker combination to obtain data for determining a subject's risk of developing one or more CV events, the CVD marker combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising assaying a sample from the subject to determine a concentration of at least one Cer of formula I and at least one PC of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

[0041] In certain embodiments of the use of a CVD marker combination to obtain data as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II, and the CVD marker combination includes at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to a control indicates that the subject is at risk of developing one or more cardiovascular events, and the CVD marker combination further comprises at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1.

[0042] In another aspect of the present disclosure, there is provided a use of a CVD marker combination for assessing the effectiveness of a CVD treatment in a subject, the CVD marker combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising assaying a sample from the subject to determine the concentration of the at least one ceramide (Cer) of formula I and the concentration of the at least one phosphatidylcholine (PC) of formula II, wherein a decreased concentration of at least one Cer of formula I and / or an increased concentration of at least one PC of formula II in the sample compared to a control indicates the effectiveness of said treatment.

[0043] In certain embodiments of the use of a CVD marker combination to assess the effectiveness of a CVD treatment as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II, and the CVD marker combination comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC compared to a control indicates efficacy of said treatment, and the CVD marker combination further comprises at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1.

[0044] In another aspect of the present disclosure, there is provided a use of a combination of CVD markers for selecting an appropriate CVD treatment for a subject, the CVD marker combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising assaying a sample from the subject to determine the concentration of at least one Cer of formula I and at least one PC of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject requires treatment, or a modification or supplementation of an already administered treatment.

[0045] In certain embodiments of the use of a CVD marker combination to select an appropriate CVD treatment as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II, and the CVD marker combination comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC as set forth in Table 1 compared to a control indicates that the subject requires treatment, or a change or supplementation of an already administered treatment, and the CVD marker combination further comprises at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1.

[0046] In another aspect of the present disclosure, there is provided a standard combination for determining the risk of a subject developing one or more cardiovascular events, the standard combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II. In certain embodiments, the standard combination comprises at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II. In certain embodiments, the isotopes of at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II are deuterium, 13 C or 15 It's N.

[0047] In certain embodiments of the standard combination as described herein, at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

[0048] In certain embodiments of the standard combination as described herein, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cer of formula I are isotopically labeled Cer of formula I, and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PC of formula II are isotopically labeled PC of formula II. In certain embodiments, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotope(s) of isotopically labeled Cer of formula I and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotope(s) of isotopically labeled PC of formula II are deuterium, 13 C or 15 It's N. In certain embodiments, the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1. In certain embodiments, at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 is isotopically labeled. In certain embodiments, the isotope of at least one additional isotopically labeled Cer and / or at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1 is deuterium, 13 C or 15 It's N.

[0049] In another aspect of the present disclosure, in the preparation of a reagent, kit or composition, there is provided a standard combination for assaying the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample from a subject to determine the risk of the subject developing one or more cardiovascular events, the standard combination comprising at least one Cer of formula I and at least one PC of formula II.

[0050] In certain embodiments of a standard combination for assaying as described herein, at least one Cer of formula I and / or at least one PC of formula II is isotopically labeled. In certain embodiments, the isotopes of at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II are deuterium, 13 C or 15 It's N. In certain embodiments of a standard combination for assaying as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

[0051] In certain embodiments of a standard combination for an assay as described herein, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cer of formula I are isotopically labeled and / or at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PC of formula II are isotopically labeled. In certain embodiments, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotope-labeled isotopes of Cer of formula I and / or at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotope-labeled isotopes of PC of formula II are deuterium, 13 C or 15 It's N. In certain embodiments, the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1. In certain embodiments, at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 is isotopically labeled. In certain embodiments, the isotope of at least one additional isotopically labeled Cer and / or the isotope of at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1 is deuterium, 13 C or 15 It's N.

[0052] In another aspect of the present disclosure, there is provided a standard combination for assaying the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample from a subject in the preparation of a reagent, kit or composition for evaluating the effectiveness of a CVD treatment in a subject, the standard combination comprising at least one Cer of formula I and at least one PC of formula II.

[0053] In certain embodiments of a standard combination for assessing efficacy as described herein, at least one Cer of formula I and / or at least one PC of formula II is isotopically labeled. In certain embodiments, the isotopes of at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II are deuterium, 13 C or 15 It's N. In certain embodiments of a standard combination for assessing efficacy as assayed as described herein, at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or at least one PC of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II.

[0054] In certain embodiments of a standard combination for assaying and assessing efficacy as described herein, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are isotopically labeled and / or at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are isotopically labeled. In certain embodiments, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotope(s) of isotopically labeled Cer of formula I and / or at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotope(s) of isotopically labeled PC of formula II are deuterium, 13 C or 15 It's N. In certain embodiments, the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1. In certain embodiments, at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 is isotopically labeled. In certain embodiments, the isotope of at least one additional isotopically labeled Cer and / or at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1 is deuterium, 13 C or 15 It's N.

[0055] In another embodiment of the present disclosure, there is provided a standard combination for assaying the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample from a subject in the preparation of a reagent, kit or composition for selecting an appropriate CVD treatment for the subject, the standard combination comprising at least one Cer of formula I and at least one PC of formula II. In certain embodiments of a standard combination for assaying as described herein and selecting an appropriate CVD treatment, at least one Cer of formula I and / or at least one PC of formula II is isotopically labeled. In certain embodiments, the isotope of at least one isotopically labeled Cer of formula I and / or the isotope of at least one isotopically labeled PC of formula II is deuterium, 13 C or 15 It's N.

[0056] In certain embodiments of the standard combination for selecting an appropriate CVD treatment, assayed as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II.

[0057] In certain embodiments of a standard combination for assaying as described herein and selecting an appropriate CVD treatment, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are isotopically labeled and / or at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are isotopically labeled. In certain embodiments, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotope(s) of isotopically labeled Cer of formula I and / or at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotope(s) of isotopically labeled PC of formula II are deuterium, 13 C or 15 It's N. In certain embodiments, the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1. In certain embodiments, at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 is isotopically labeled. In certain embodiments, the isotope of at least one additional isotopically labeled Cer and / or at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1 is deuterium, 13 C or 15 It's N.

[0058] In another aspect of the present disclosure, there is provided a use of a standard combination for determining a subject's risk of developing one or more cardiovascular events, the standard combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising assaying a sample from the subject to determine the concentration of at least one Cer of formula I and at least one PC of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject has a risk of developing one or more cardiovascular events.

[0059] In certain embodiments of the use of the standard combination for determining risk as described herein, at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II, and the standard combination comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use of the standard combination further comprises assaying the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, where an increased or decreased concentration of at least one additional Cer and / or at least one additional PC shown in Table 1 compared to a control indicates that the subject is at risk of developing one or more cardiovascular events, and the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

[0060] In another aspect of the present disclosure, there is provided a use of a standard combination for detecting the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample from a subject, the standard combination comprising at least one Cer of formula I and at least one PC of formula II.

[0061] In certain embodiments of the use of the standard combination for detection as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II, and the standard combination comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use further comprises detecting the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 in the sample, and the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1. In certain embodiments, the use comprises detecting an altered concentration of at least one Cer of formula I and / or an altered concentration of at least one PC of formula II in a sample from the subject. In certain embodiments, the use comprises detecting an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in a sample from a subject.

[0062] In another aspect of the present disclosure, there is provided a use of a standard combination for obtaining data for determining a subject's risk of developing one or more CV events, the standard combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising assaying a sample from the subject to determine the concentration of at least one Cer of formula I and at least one PC of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject has a risk of developing one or more cardiovascular events.

[0063] In certain embodiments of the use of the standard combination to obtain data as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II, and the standard combination comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to a control indicates that the subject is at risk of developing one or more cardiovascular events, and the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

[0064] In another aspect of the present disclosure, there is provided a use of a standard combination for assessing the effectiveness of a CVD treatment in a subject, the standard combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising assaying a sample from the subject to determine the concentration of the at least one Cer of formula I and the concentration of the at least one PC of formula II, wherein a decreased concentration of the at least one Cer of formula I and / or an increased concentration of the at least one PC of formula II in the sample compared to a control indicates the effectiveness of said treatment.

[0065] In certain embodiments of the use of the standard combination to evaluate the effectiveness of a CVD treatment as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II, wherein the standard combination comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC compared to the control indicates efficacy of said treatment, and the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

[0066] In another embodiment of the present disclosure, there is provided a use of a standard combination for selecting an appropriate CVD treatment for a subject, the standard combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising assaying a sample from the subject to determine the concentration of the at least one ceramide (Cer) of formula I and the concentration of the at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject requires treatment, or a modification or supplementation of an already administered treatment.

[0067] In certain embodiments of the use of the standard combination to select a suitable CVD treatment as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II, wherein the standard combination comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC as set forth in Table 1 compared to the control indicates that the subject requires treatment, or a change or supplementation of an already administered treatment, and the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

[0068] In certain embodiments of all aspects of the present disclosure, the standard combination comprises at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II. In certain embodiments of all aspects of the disclosure, the isotopes of the at least one isotopically labeled Cer of formula I and / or the at least one isotopically labeled PC of formula II are deuterium, 13C or 15 It's N. In certain embodiments of all aspects of the present disclosure, the standard combination comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 isotopically labeled Cer of formula I and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 isotopically labeled PC of formula II. In certain embodiments of all aspects of the disclosure, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotope(s) of Cer of formula I and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotope(s) of PC of formula II are deuterium, 13 C or 15 It's N.

[0069] In certain embodiments of all aspects of the present disclosure, at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 is isotopically labeled. In certain embodiments of all aspects of the present disclosure, the isotope of the at least one additional isotopically labeled Cer and / or the at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1 is deuterium, 13 C or 15 It's N.

[0070] In certain embodiments of all aspects of the present disclosure, the standard combination comprises at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled Cer and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled PC selected from the following isotopically labeled lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:0), Cer(d18:1 / 26:1), Cer(d16:1 / 1 6:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(d18: 2 / 24:1), Cer(d18:2 / 24:0), Cer(d20:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6, PC16:0 / 16:0, PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:8. In certain embodiments of all aspects of the present disclosure, at least one, at least two, at least three, at least four, at least five, at least six, or at least seven isotopically labeled lipids of the standard combination are selected from the following isotopically labeled lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6, and PC16:0 / 16:0. In certain embodiments of all aspects of the present disclosure, the standard combinations of isotopically labeled lipids are Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and PC16:0 / 16:0. In certain embodiments of all aspects of the present disclosure, the standard combination of isotopically labeled lipids Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and / or PC16:0 / 16:0 are deuterium labeled. In certain embodiments of all aspects of the present disclosure, the standard combination of isotopically labeled lipids is d7-Cer(d18:1 / 16:0), d7-Cer(d18:1 / 18:0), d7-Cer(d18:1 / 24:0), d7-Cer(d18:1 / 24:1), d9-PC16:0 / 22:5, d9-PC14:0 / 22:6 and / or d9-PC16:0 / 16:0.

[0071] In another aspect of the present disclosure, there is provided a use of one or more reagents in the preparation of a reagent, kit or composition for determining a subject's risk of developing one or more cardiovascular events, comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

[0072] In certain embodiments of the use of one or more reagents for determining risk as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II. In certain embodiments, the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

[0073] In another aspect of the present disclosure, there is provided the use of one or more reagents in the preparation of a reagent, kit or composition for detecting the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample from a subject.

[0074] In certain embodiments of the use of one or more reagents for detection as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II. In certain embodiments, the use further comprises detecting the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1 in the sample. In certain embodiments, the use of one or more reagents for detection as described herein comprises detecting an altered concentration of at least one Cer of formula I and / or an altered concentration of at least one PC of formula II in a sample from a subject. In certain embodiments, the use comprises detecting an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in a sample from a subject. In certain embodiments, the use of one or more reagents in the preparation of a reagent, kit or composition for detecting the concentration of at least one ceramide of formula I and at least one phosphatidylcholine of formula II in a sample from a subject comprises using a standard combination comprising at least one Cer of formula I and / or at least one PC of formula II.

[0075] In another aspect of the present disclosure, there is provided a use of one or more reagents in the preparation of a reagent, kit or composition for obtaining data for determining a subject's risk of developing one or more CV events, the use comprising assaying a sample from the subject to determine a concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

[0076] In certain embodiments of the use of one or more reagents to obtain data as described herein, at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I, and / or at least one PC of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II. In certain embodiments, the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

[0077] In another aspect of the present disclosure, there is provided a use of one or more reagents in the preparation of a reagent, kit or composition for assessing the effectiveness of a CVD treatment in a subject, the use comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein a decreased concentration of at least one Cer of formula I and / or an increased concentration of at least one PC of formula II in the sample compared to a control indicates the effectiveness of said treatment.

[0078] In certain embodiments of the use of one or more reagents for assessing the effectiveness of a CVD treatment as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC compared to a control indicates the effectiveness of said treatment.

[0079] In another aspect of the present disclosure, there is provided the use of one or more reagents in the preparation of a reagent, kit or composition for selecting an appropriate CVD treatment for a subject, the method comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject requires treatment, or a modification or supplementation of an already administered treatment.

[0080] In certain embodiments of the use of one or more reagents for selecting a suitable CVD treatment as described herein, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC as set forth in Table 1 compared to a control indicates that the subject requires treatment, or a modification or supplementation of an already administered treatment. In certain embodiments, the reagent comprises any standard, control, substance, compound, solution, solvent, agent, component, preparation, or any combination thereof used for the disclosed methods and uses. In certain embodiments, the reagent is a combination or mixture of any standard, control, substance, compound, solution, solvent, agent, component, and preparation used for the disclosed methods and uses.

[0081] In some embodiments, one or more of any of the components of the reagent is isotopically labeled. In some embodiments, the isotope of the isotope of the isotopically labeled component is deuterium, 13 C or 15 It's N.

[0082] In certain embodiments of all aspects of the present disclosure, the reagent comprises at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled Cer and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled PC selected from the following isotopically labeled lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:0), Cer(d18:1 / 26:1), Cer(d16:1 / 16: 0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(d18:2 / 24:1), Cer(d18:2 / 24:0), Cer(d20:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6, PC16:0 / 16:0, PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:8.

[0083] In certain embodiments of all aspects of the present disclosure, at least one, at least two, at least three, at least four, at least five, at least six, or at least seven isotopically labeled lipids of the reagents are selected from the following isotopically labeled lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6, and PC16:0 / 16:0.

[0084] In certain embodiments of all aspects of the present disclosure, the isotopically labeled lipids of the reagent are Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and PC16:0 / 16:0. In certain embodiments of all aspects of the disclosure, the isotopically labeled lipids of the reagent Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and / or PC16:0 / 16:0 are deuterium labeled. In certain embodiments of all aspects of the present disclosure, the isotopically labeled lipid of the reagent is d7-Cer(d18:1 / 16:0), d7-Cer(d18:1 / 18:0), d7-Cer(d18:1 / 24:0), d7-Cer(d18:1 / 24:1), d9-PC16:0 / 22:5, d9-PC14:0 / 22:6 and / or d9-PC16:0 / 16:0. In certain embodiments, the reagent is used to assay a sample to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II. In some embodiments, the reagent is further used to assay a sample to determine the concentration of at least one additional Cer and / or PC selected from any of the Cer and PC species mentioned in Table 1 of the present disclosure. In certain embodiments, the reagents are used in the preparation of reagents, kits, or compositions used to practice the methods and uses of the present disclosure.

[0085] In another aspect of the present disclosure, there is provided a composition or kit for diagnosing, predicting or detecting one or more cardiovascular events in a subject or for carrying out any of the methods or uses according to the present disclosure, comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II.

[0086] In certain embodiments of the composition or kit, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II. In certain embodiments, the composition or kit further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1. In certain embodiments, the composition or kit comprises at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II. In certain embodiments, the isotopes of at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II are deuterium, 13 C or 15 It's N. In certain embodiments of the composition or kit, the at least one isotopically labeled Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 isotopically labeled Cer of formula I and / or the at least one isotopically labeled PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 isotopically labeled PC of formula II. In certain embodiments, the isotopes of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled Cer of formula I and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled PC of formula II are deuterium, 13 C or 15 It's N. In certain embodiments, the composition or kit comprises at least one additional isotopically labeled Cer and / or at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1. In certain embodiments, the isotope of at least one additional isotopically labeled Cer and / or at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1 is deuterium, 13 C or 15 It's N.

[0087] In certain embodiments of all aspects of the present disclosure, the composition or kit comprises at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled Cer and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled PC selected from the following isotopically labeled lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:0), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(d18 :2 / 24:1), Cer(d18:2 / 24:0), Cer(d20:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6, PC16:0 / 16:0, PC18:0 / 20:5, PC16: 0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:8.

[0088] In certain embodiments of all aspects of the present disclosure, at least one, at least two, at least three, at least four, at least five, at least six, or at least seven isotopically labeled lipids of the composition or kit are selected from the following isotopically labeled lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6, and PC16:0 / 16:0. In certain embodiments of all aspects of the present disclosure, the isotopically labeled lipids of the composition or kit are Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and PC16:0 / 16:0.

[0089] In certain embodiments of all aspects of the present disclosure, the isotopically labeled lipids Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and / or PC16:0 / 16:0 of ​​the composition or kit are deuterium labeled. In certain embodiments of all aspects of the present disclosure, the isotopically labeled lipid of the composition or kit is d7-Cer(d18:1 / 16:0), d7-Cer(d18:1 / 18:0), d7-Cer(d18:1 / 24:0), d7-Cer(d18:1 / 24:1), d9-PC16:0 / 22:5, d9-PC14:0 / 22:6 and / or d9-PC16:0 / 16:0. The composition or kit may further comprise standards, controls, reagents, solutions, solvents, containers, instructions for use and / or other elements for carrying out the methods or uses disclosed herein.

[0090] In some embodiments, the composition or kit includes elements for collecting a blood sample, for example, a dried blood spot on a filter. The composition or kit may be a test kit for laboratory use or a home test kit (commercial test). The composition or kit may be any combination of standards, controls, reagents, solutions or solvents purchased from commercial manufacturers or prepared in-house laboratories, used to diagnose, predict or detect one or more cardiovascular events in a subject, or to carry out any of the methods or uses according to the present disclosure. In certain embodiments, the composition or kit is a combination of any standard, control, reagent, solution or solvent, purchased from a commercial manufacturer or prepared in an in-house laboratory, used to assay the concentration of at least one Cer of formula I and at least one PC of formula II.

[0091] In certain embodiments of a composition or kit having any combination of standards, controls, reagents, solutions or solvents purchased from a commercial manufacturer or prepared in an in-house laboratory, the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II. In certain embodiments, the composition or kit is a combination of any standard, control, reagent, solution or solvent, purchased from a commercial manufacturer or prepared in an in-house laboratory, used to assay the concentration of at least one Cer and / or at least one PC selected from any of the Cer and PC species mentioned in Table 1. The composition or kit can be used for assays carried out using various chemical and high-resolution analytical techniques as required. Suitable analytical techniques according to the method and use of the present invention include, but are not limited to, mass spectrometry (MS) and nuclear magnetic resonance (NMR). Any high-resolution technique capable of resolving individual biomarkers can be used to collect information about the concentration of the biomarker in question, for example, a biomarker profile from a biological sample. Typically, the information is collected using mass spectrometry. MS analysis can be coupled with another high-performance separation method, for example, gas chromatography (GC), two-dimensional gas chromatography (GCxGC), liquid chromatography (LC), two-dimensional liquid chromatography (LCxLC), high performance liquid chromatography (HPLC) or ultra-high performance liquid chromatography (UHPLC).

[0092] In another aspect of the disclosure, there is provided a method for monitoring the progression of a subject's risk of a CVD and / or CV event, comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, and comparing these concentrations with the concentrations in a sample previously obtained from the same subject. In certain embodiments of the methods for monitoring the progression of a subject's risk of a CVD and / or CV event as described herein, at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, and / or at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II.

[0093] In certain embodiments, the method further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1. In certain embodiments of all aspects of the present disclosure, the methods and uses further comprise the step of adding at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II to the sample prior to assaying the concentration of at least one Cer of formula I and / or at least one PC of formula II. The at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II may be, but is not limited to, a deuterium-labeled Cer of formula I and / or a deuterium-labeled PC of formula II. The at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II may also be 13 C label or 15 In certain embodiments of all aspects of the present disclosure, the methods and uses further comprise adding at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled Cer of formula I and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled PC of formula II to the sample. The at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled Cer of formula I and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten isotopically labeled PC of formula II may be, but is not limited to, deuterium labeled, 13 C-labeled or 15It may be N-labeled Cer of formula I and / or deuterium-labeled PC of formula II.

[0094] In certain embodiments of all aspects of the present disclosure, the methods and uses further comprise the step of adding to the sample at least one additional isotopically labeled Cer and / or at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1. The at least one additional isotopically labeled Cer and / or at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1 includes, but is not limited to, deuterium-labeled Cer and / or at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1. 13 C-labeled or 15 N-labeled Cer, and / or deuterium-labeled, 13 C-labeled or 15 N-labeled PC.

[0095] In certain embodiments of all aspects of the present disclosure, the isotopically labeled Cer and / or PC is selected from the following isotopically labeled lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:0), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(d18 :2 / 24:1), Cer(d18:2 / 24:0), Cer(d20:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6, PC16: 0 / 16:0, PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16 :1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:8.

[0096] In certain embodiments of all aspects of the present disclosure, the isotopically labeled Cer and / or PC is selected from the following isotopically labeled lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and PC16:0 / 16:0. In certain embodiments of all aspects of the present disclosure, the isotopically labeled Cer and PC are Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and PC16:0 / 16:0. In certain embodiments of all aspects of the disclosure, the isotopically labeled Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and / or PC16:0 / 16:0 are deuterium labeled. In certain embodiments of all aspects of the present disclosure, the isotopically labeled lipid is d7-Cer(d18:1 / 16:0), d7-Cer(d18:1 / 18:0), d7-Cer(d18:1 / 24:0), d7-Cer(d18:1 / 24:1), d9-PC16:0 / 22:5, d9-PC14:0 / 22:6 and / or d9-PC16:0 / 16:0.

[0097] In certain embodiments of all aspects of the present disclosure, the concentration of at least one Cer of formula I and the concentration of at least one PC of formula II are determined in one assay. In some embodiments, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are determined in the same assay. In some embodiments, the concentrations of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 are determined in the same assay. In some embodiments, one assay is performed by mass spectrometry (MS). In certain embodiments of all aspects of the present disclosure, the at least one additional Cer is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional Cers selected from any of the Cer species mentioned in Table 1. In some embodiments, the at least one additional Cer is a combination including, but not limited to, a Cer / Cer ratio of at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional Cers. The combination may also be another combination described herein.

[0098] In certain embodiments of all aspects of the present disclosure, the at least one additional PC is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional PCs selected from any of the PC species mentioned in Table 1. In some embodiments, the at least one additional PC is a combination including, but not limited to, a PC / PC ratio of at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional PCs. The combination may also be another combination described herein.

[0099] In certain embodiments of all aspects of the disclosure, the method or use further comprises determining the level of total cholesterol (TC), low density lipoprotein (LDL), low density lipoprotein cholesterol (LDL-C), high density lipoprotein (HDL), high density lipoprotein cholesterol (HDL-C), low density lipoprotein particle (LDL-P), high density lipoprotein particle (HDL-P), apolipoprotein B (ApoB), apolipoprotein AI (ApoAI), apolipoprotein AII (ApoAII), and / or apolipoprotein C (ApoC) in a sample from the subject. In certain embodiments, the subject does not have an elevated level of one or more of total cholesterol, low density lipoprotein cholesterol (LDL-C), apolipoprotein C (ApoC) or apolipoprotein B (ApoB), or a decreased level of HDL-cholesterol (HDL-C). In certain embodiments of all aspects of the disclosure, the method or use further comprises determining other cardiovascular biomarkers, including, but not limited to, triglycerides (TG), C-reactive protein (CRP), troponin T (TNT or TnT), troponin I (TNI or TnI), B-type natriuretic peptide (BNP), N-terminal pro-B-type natriuretic peptide (NT-proBNP), cystatin C, glycated hemoglobin A1c (HbA1c), glucose, tumor suppressor 2 (St2), galectin, trimethylamine-N-oxide (TMAO), lipoprotein-associated phospholipase A2 (Lp-PLA2), growth differentiation factor 15 (GDF15), lipoprotein(a) (Lp(a)), any other lipoprotein subgroup composition or particle number, or any combination thereof.

[0100] In certain embodiments of all aspects of the present disclosure, the method or use may further comprise determining any other cardiovascular or metabolic biomarker, including but not limited to creatine kinase (CK). In certain embodiments of all aspects of the present disclosure, the method or use further comprises using other personal or health data, such as gender, age, blood pressure, BMI, smoking status, diabetes, lipid-lowering treatment or other medications, history of cardiovascular events, ethnic minority background, geographic location, medical imaging data, such as angiographic or computed tomography (CT) medical imaging data, or any combination thereof.

[0101] In certain embodiments of all aspects of the present disclosure, the method or use may further include using any personal information or health data including, but not limited to, medical history of CVD and / or CV events, family history of CVD and / or CV events, family history of diabetes, family history of other diseases. In certain embodiments of all aspects of the present disclosure, cardiovascular (CV) events include stable angina, unstable angina, myocardial infarction (MI), acute myocardial infarction (AMI), acute coronary syndrome (ACS), stroke, transient ischemic attack, deep vein thrombosis, heart failure, hospitalization for heart failure, and cardiovascular death. In certain embodiments of all aspects of the present disclosure, the cardiovascular (CV) event comprises peripheral arterial disease (also called peripheral arterial disease) and / or arrhythmia, e.g., atrial fibrillation.

[0102] In certain embodiments of all aspects of the present disclosure, the subject is any mammalian subject for which prediction, diagnosis or treatment is desired. In certain embodiments, the subject is a human. In some embodiments, the subject is a healthy individual with no prior signs or symptoms of CVD. In other embodiments, the subject has previously suffered or is suffering from CVD or CAD. The subject may also have previously suffered from a cardiovascular event, such as angina, myocardial infarction or stroke. In some embodiments, the subject suffers from a metabolic disease, such as diabetes. In some embodiments, the subject is suspected of suffering from CVD and / or is at high risk of developing a CV event. In some embodiments, the subject has normal levels of traditional risk markers, such as total cholesterol, LDL or HDL, and / or does not have traditional risk factors, such as diabetes, hypertension or smoking. In certain embodiments, the subject is undergoing treatment, such as statin treatment.

[0103] In certain embodiments of all aspects of the present disclosure, the sample is a biological sample obtained from a subject or a group or population of subjects.In certain embodiments, the sample is a blood sample, a serum sample, a plasma sample, a saliva sample, a urine sample, a tissue sample or a fraction thereof, such as a lipoprotein fraction.In certain embodiments, the sample is a serum or plasma sample.The sample can be prepared using techniques well known in the art.In certain embodiments, the blood sample is a dried blood spot.In certain embodiments, the blood sample is dried on a filter.

[0104] In certain embodiments of all aspects of the present disclosure, the control is a control sample or control value obtained from a single healthy individual or a general population of healthy individuals.In some embodiments, the control is a control value or a set of data for biomarkers in a sample that has been previously determined.In some embodiments, the control sample is a control value obtained from the same subject previously, or from a sample previously collected from the same subject, for example, for use in controlling the progression of CVD. In certain embodiments, the control is a CVD patient who has not yet experienced any major CV event, or is from a group of CVD patients who have not yet experienced any major CV event. In certain embodiments, the control is a control value, e.g., a concentration score or combined value of at least one Cer of formula I and at least one PC of formula II, or a combined value of at least one Cer of formula I and at least one PC of formula II with at least one additional Cer or PC selected from any of the Cer and PC species mentioned in Table 1.

[0105] In certain embodiments of all aspects of the present disclosure, the treatment includes any therapeutic treatment typically given to subjects at risk of developing or suffering from CVD, including, but not limited to, administering a drug and / or providing the subject with treatment, behavioral and / or lifestyle modifications. In some embodiments, the behavioral and / or lifestyle modifications include, for example, lifestyle interventions and / or counseling, including instructions, encouragement and / or follow-up regarding healthy eating, weight management, physical activity / exercise and / or smoking cessation. In certain embodiments, the treatment includes surgery, such as angioplasty, stent placement or diversion surgery. In certain embodiments, the treatment includes controlling the progression of CVD and / or its complications.

[0106] In certain embodiments of all aspects of the present disclosure, the drug comprises any pharmaceutical agent typically given to subjects with CVD, such as a statin, another lipid-lowering / modifying drug, and / or a lipid concentration modulator. In some embodiments, the drug is an HMG-CoA reductase inhibitor (a statin or other HMG-CoA reductase inhibitor), niacin (nicotinic acid), a cholesterol absorption inhibitor (ezetimibe), a cholesteryl ester transfer protein (CETP) inhibitor, a bile acid sequestrant, a fibrate, a phytosterol, an anti-inflammatory agent (e.g., methotrexate, IL-1 mAb, TNF-alpha mAb), acetylsalicylic acid (aspirin) or a PCSK9 inhibitor, or a lipid concentration modulator selected from a small molecule, an antibody, an antisense RNA, a small interfering RNA (siRNA), and a natural or modified lipid, or any combination thereof. In certain embodiments, the statin comprises atorvastatin, cerivastatin, fluvastatin, fluvastatin XL, lovastatin, pitavastatin, pravastatin, rosuvastatin, simvastatin, or any combination thereof.

[0107] In some embodiments, the cholesterol absorption inhibitor is ezetimibe or SCH-48461, the cholesteryl ester transfer protein (CETP) inhibitor is evacetrapib, anacetrapib or dalcetrapib, the bile acid sequestrant is colesevelam, cholestyramine or colestipol, the fibrate is fenofibrate, gemfibrozil, clofibrate or bezafibrate, and the PCSK9 inhibitor is a PCSK9-specific antibody, siRNA, or peptidomimetic.

[0108] In certain embodiments of all aspects of the present disclosure, the determination of the concentration of Cer and PC is carried out using a typical assay.In certain embodiments, such assay is or includes mass spectrometry, nuclear magnetic resonance spectroscopy, fluorescence spectroscopy or dual polarization interferometry, high performance separation methods, such as liquid chromatography (LC), gas chromatography (GC), two-dimensional liquid chromatography (LCxLC), two-dimensional gas chromatography (GCxGC), high performance liquid chromatography (HPLC), ultra-high performance liquid chromatography (UHPLC) or ultra-high performance liquid chromatography (UPLC), immunoassays, such as ELISA, and / or assays using binding moieties that can specifically bind to analytes.In certain embodiments, the assay is mass spectrometry (MS).The MS device can be coupled with direct sample injection, such as robotic nanoflow ion source device, or high performance separation methods, such as HPLC, UHPLC, or UPLC.

[0109] In certain embodiments of all aspects of the disclosure, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed. In another embodiment, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed. In yet another embodiment, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed.

[0110] In certain embodiments of all aspects of the disclosure, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten additional Cers selected from any of the Cer species mentioned in Table 1 are assayed. In another embodiment, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten additional PCs selected from any of the PC species mentioned in Table 1 are assayed. In yet another embodiment, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, or at least eight, at least nine, or at least ten additional Cers of formula I selected from any of the Cer species mentioned in Table 1 and / or at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten additional PCs of formula II selected from any of the PC species mentioned in Table 1 are assayed.

[0111] In certain embodiments of all aspects of the present disclosure, R of formula I 1 is a saturated, monounsaturated or diunsaturated alkyl chain having 11 to 17 carbon atoms, R 2 is a saturated alkyl chain having 13 to 21 carbon atoms, a monounsaturated or diunsaturated alkyl chain having 13 to 25 carbon atoms. In certain embodiments of all aspects of the present disclosure, R of formula I 1 is a monounsaturated or diunsaturated alkyl chain having 13 to 15 carbon atoms, and R 2 is a saturated, monounsaturated or diunsaturated alkyl chain having 15 to 25 carbon atoms. In certain embodiments of all aspects of the present disclosure, R of formula I 1 is a monounsaturated or diunsaturated alkyl chain having 13 to 15 carbon atoms, and R2 is a saturated alkyl chain having 15 to 21 carbon atoms, a monounsaturated or diunsaturated alkyl chain having 15 to 25 carbon atoms.

[0112] In all aspects of the present disclosure, the PC may be a brutto PC or a molecular PC. In certain embodiments of all aspects of the present disclosure, R of formula II 3 and R 4 is a saturated, monounsaturated, or polyunsaturated alkyl chain having 11 to 26 carbon atoms. In certain embodiments of all aspects of the present disclosure, R 3 and R 4 is a saturated, monounsaturated or polyunsaturated alkyl chain having up to six double bonds.

[0113] In certain embodiments of all aspects of the disclosure, the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cer of formula I is assayed, including, but not limited to, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), ... :0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(d18:2 / 24:1) and / or Cer(d20:1 / 24:1). In some embodiments, the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including, but not limited to, PC16:0 / 22:5, PC14:0 / 22:6, PC18:0 / 20:5, PC16:0 / 20:4, PC 18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:8.In other embodiments, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including, but not limited to, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(d18:2 / 24:1), and / or Cer(d20:1 / 24:1). The concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including: PC16:0 / 22:5, PC14:0 / 22:6, PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, P C18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:8.

[0114] In certain embodiments of all aspects of the disclosure, the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1). In some embodiments, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including, but not limited to, PC16:0 / 22:5, PC14:0 / 22:6, PC16:0 / 22:6, PC16:0 / 20:4, PC18:0 / 20:4, PC17:0 / 20:3, PC36:8, PC36:6, PC36:4, PC38:4, PC38:7, PC34:4, and / or PC40:8. In other embodiments, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including, but not limited to, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1), including, but not limited to, at least one, at least two, at least one The concentrations of at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten PCs of formula II are assayed: PC16:0 / 22:5, PC14:0 / 22:6, PC16:0 / 22:6, PC16:0 / 20:4, PC18:0 / 20:4, PC17:0 / 20:3, PC36:8, PC36:6, PC36:4, PC38:4, PC38:7, PC34:4 and / or PC40:8.

[0115] In certain embodiments of all aspects of the disclosure, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1). In some embodiments, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including but not limited to: PC16:0 / 22:5 and / or PC14:0 / 22:6. In other embodiments, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0) and / or Cer(d18:1 / 24:1); and at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including but not limited to: PC16:0 / 22:5 and / or PC14:0 / 22:6. In certain embodiments of all aspects of the present disclosure, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0) and / or Cer(d18:1 / 24:1), at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 are assayed.

[0116] In certain embodiments of all aspects of the present disclosure, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including but not limited to: PC16:0 / 22:5 and / or PC14:0 / 22:6, the concentrations of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 are assayed.

[0117] In certain embodiments of all aspects of the present disclosure, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0) and / or Cer(d18:1 / 24:1); at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including but not limited to: PC16:0 / 22:5 and / or PC14:0 / 22:6; and at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 are assayed.

[0118] In certain embodiments of all aspects of the disclosure, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including, but not limited to, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1), and the concentration of at least one additional Cer is assayed, including, but not limited to, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1). Will be: Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 26:0), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), C er(d16:1 / 18:0), Cer(d16:1 / 24:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(18:2 / 24:0), Cer(d18:2 / 24:1), Cer(d20 :1 / 24:0) and / or Cer (d20:1 / 24:1. In some embodiments, the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including, but not limited to, PC16:0 / 22:5 and / or PC14:0 / 22:6, and the concentration of at least one additional PC is assayed, including, but not limited to, PC16: :0 / 16:0, 16:0 / 22:5, PC14:0 / 22:6, PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:8.In other embodiments, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including, but not limited to, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1), including, but not limited to, at least one, at least two, at least three, at least four, The concentrations of at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed: PC16:0 / 22:5 and / or PC14:0 / 22:6, and the concentration of at least one additional Cer is assayed, including, but not limited to, Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 26:0), Cer(d18 :1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(18:2 / 24:0), Cer(d18:2 / 24:1), Cer(d20:1 / 24:0) and / or Cer(d20:1 / 24:1), and the concentration of at least one additional PC is assayed, including, but not limited to, PC16:0 / 16:0 , 16:0 / 22:5, PC14:0 / 22:6, PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:8.

[0119] In certain embodiments of all aspects of the disclosure, the concentration of Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1) and / or Cer(d18:1 / 24:0) is assayed. In some embodiments, the concentration of PC16:0 / 22:5 and / or PC14:0 / 22:6 is assayed.

[0120] In certain embodiments of all aspects of the disclosure, the concentrations of at least two, at least three, at least four, at least five, or at least six of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, and PC14:0 / 22:6.

[0121] In other embodiments, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5 and PC14:0 / 22:6 are assayed.

[0122] In certain embodiments of all aspects of the present disclosure, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1), and the concentrations of at least one additional Cer are assayed, including but not limited to: Cer(d18:1 / 24:0). In some embodiments, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including but not limited to: PC16:0 / 22:5 and / or PC16:0 / 22:6. In other embodiments, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1); at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including but not limited to: PC16:0 / 22:5 and / or PC16:0 / 22:6; and at least one additional Cers are assayed, including but not limited to: Cer(d18:1 / 24:0).

[0123] In certain embodiments of all aspects of the disclosure, the concentrations of Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1) and / or Cer(d18:1 / 24:0) are assayed. In some embodiments, the concentrations of PC16:0 / 22:5, PC14:0 / 22:6 and / or PC16:0 / 16:0 are assayed. In certain embodiments of all aspects of the disclosure, the concentrations of at least two, at least three, at least four, at least five, at least six, or at least seven of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC14:0 / 22:6, and PC16:0 / 16:0. In other embodiments, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC14:0 / 22:6 and PC16:0 / 16:0 are assayed.

[0124] In certain embodiments of all aspects of the present disclosure, the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1), and the concentration of at least one additional Cers is assayed, including but not limited to: Cer(d18:1 / 24:0). In some embodiments, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including but not limited to: PC16:0 / 22:5 and / or PC16:0 / 22:6, and the concentration of at least one additional PC is assayed, including but not limited to: PC16:0 / 16:0. In other embodiments, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0) and / or Cer(d18:1 / 24:1); at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including but not limited to: PC16:0 / 22:5 and / or PC16:0 / 22:6; at least one additional Ce is assayed, including but not limited to: Cer(d18:1 / 24:0); at least one additional PC is assayed, including but not limited to: PC16:0 / 16:0.

[0125] In certain embodiments of all aspects of the disclosure, the concentrations of Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1) and / or Cer(d18:1 / 24:0) are assayed. In some embodiments, the concentrations of PC16:0 / 22:5, PC16:0 / 22:6 and / or PC16:0 / 16:0 are assayed.

[0126] In certain embodiments of all aspects of the disclosure, the concentrations of at least two, at least three, at least four, at least five, at least six, or at least seven of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC16:0 / 22:6, and PC16:0 / 16:0. In other embodiments, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC16:0 / 22:6 and PC16:0 / 16:0 are assayed.

[0127] In certain embodiments of all aspects of the present disclosure, the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1), and the concentration of at least one additional Cers is assayed, including but not limited to: Cer(d18:1 / 24:0). In some embodiments, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including but not limited to: PC16:0 / 22:5 and / or PC40:8, and the concentration of at least one additional PC is assayed, including but not limited to: PC16:0 / 18:3. In other embodiments, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0) and / or Cer(d18:1 / 24:1); at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including but not limited to: PC16:0 / 22:5 and / or PC40:8; at least one additional Ce is assayed, including but not limited to: Cer(d18:1 / 24:0); at least one additional PC is assayed, including but not limited to: PC16:0 / 18:3.

[0128] In certain embodiments of all aspects of the disclosure, the concentrations of at least Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1) and Cer(d18:1 / 24:0) are assayed. In some embodiments, the concentrations of at least PC16:0 / 22:5, PC40:8 and PC16:0 / 18:3 are assayed.

[0129] In certain embodiments of all aspects of the disclosure, the concentrations of at least two, at least three, at least four, at least five, at least six, or at least seven of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC16:0 / 22:6, and PC16:0 / 18:3. In other embodiments, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC16:0 / 22:6 and PC16:0 / 18:3 are assayed.

[0130] In certain embodiments of all aspects of the disclosure, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including, but not limited to, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1). In some embodiments, the concentrations of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including, but not limited to, PC16:0 / 20:4, PC18:0 / 20:4, and / or PC38:7. In other embodiments, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cers of formula I are assayed, including but not limited to: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), and / or Cer(d18:1 / 24:1); and at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PCs of formula II are assayed, including but not limited to: PC16:0 / 20:4, PC18:0 / 20:4, and / or PC38:7. In certain embodiments of all aspects of the disclosure, the concentrations of at least Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1) are assayed. In some embodiments, the concentrations of at least PC16:0 / 20:4, PC18:0 / 20:4, and / or PC38:7 are assayed.

[0131] In certain embodiments of all aspects of the disclosure, the concentrations of at least two, at least three, at least four, at least five, or at least six of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), PC16:0 / 20:4, PC18:0 / 20:4, and PC38:7. In other embodiments, Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), PC16:0 / 20:4, PC18:0 / 20:4 and PC38:7 are assayed.

[0132] In certain embodiments of all aspects of the present disclosure, the concentration ratio is calculated from the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cer of formula I and / or the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PC of formula II.

[0133] In certain embodiments of all aspects of the present disclosure, the concentration ratio is calculated from the concentration of at least one Cer of formula I and / or the concentration of at least one PC of formula II. In some embodiments, the concentration ratio is calculated from the concentration of Cer of formula I and the concentration of PC of formula II. In certain embodiments, the concentration ratio is Cer / Cer, Cer / PC, PC / Cer and / or PC / PC ratio, such as Cer(d18:1 / 16:0) / PC16:0 / 22:5, Cer(d18:1 / 18:0) / PC14:0 / 22:6, PC16:0 / 22:5 / Cer(d18:1 / 16:0) and / or PC14:0 / 22:6 / Cer(d18:1 / 18:0). In certain embodiments of all aspects of the present disclosure, the concentration ratio is calculated from the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1. In certain embodiments, the concentration ratio is a Cer / Cer, Cer / PC, PC / Cer and / or PC / PC ratio, e.g., Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0) and / or Cer(d18:1 / 24:0) / Cer(d18:1 / 24:1).

[0134] In certain embodiments of all aspects of the present disclosure, an increased concentration ratio of at least one Cer of formula I and at least one PC of formula II in a sample from a subject compared to a control indicates that the subject is at risk of developing one or more cardiovascular events. In certain embodiments of all aspects of the present disclosure, an increased or decreased concentration ratio of at least one additional Cer of formula I and / or at least one additional PC of formula II selected from any of the Cer and PC species mentioned in Table 1 in a sample from a subject compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

[0135] In certain embodiments of all aspects of the present disclosure, the concentration ratio is Cer / Cer, Cer / PC, PC / Cer and / or PC / PC ratio, and an increased or decreased concentration ratio in a sample from a subject compared to a control indicates that the subject is at risk for developing one or more cardiovascular events. In a typical embodiment of all aspects of the present disclosure, the following concentration ratios are determined from Cer of formula I and PC of formula II: Cer(d18:1 / 16:0) / PC16:0 / 22:5 and / or Cer(d18:1 / 18:0) / PC14:0 / 22:6. In this typical embodiment, the following concentration ratios from additional Cer species from Table 1 are also determined: Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0). In this typical embodiment of all aspects of the present disclosure, the following concentration ratios from additional PC species from Table 1 are also determined: PC16:0 / 16:0.

[0136] In another exemplary embodiment of all aspects of the present disclosure, the following concentration ratios are determined from Cer of formula I and PC of formula II: Cer(d18:1 / 16:0) / PC16:0 / 22:5 and / or Cer(d18:1 / 18:0) / PC14:0 / 22:6. In this exemplary embodiment, the following concentration ratios from additional Cer species of Table 1 are also determined: Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0).

[0137] In certain embodiments of all aspects of the present disclosure, the concentration ratio is calculated from the concentration of at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional Cers and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional PCs selected from any of the Cer and PC species mentioned in Table 1. In some embodiments, the concentration ratio comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional Cers selected from any of the Cer species mentioned in Table 1. In some embodiments, the concentration ratio comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional PCs selected from any of the PC species mentioned in Table 1. In some embodiments, the concentration ratio comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 additional Cer selected from any of the Cer species mentioned in Table 1 and at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 additional PC selected from any of the PC species mentioned in Table 1.

[0138] In certain embodiments of all aspects of the present disclosure, the concentration ratio is calculated from the concentration of at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 Cers of formula I, the concentration of at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional Cers selected from any of the Cer species mentioned in Table 1, and / or at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional PCs selected from any of the PC species mentioned in Table 1. In certain embodiments of all aspects of the present disclosure, the concentration ratio is selected from any of the lipid ratios mentioned in Tables 2, 3, 4, 6, 7, 8, 9, 10, 11, or 12.

[0139] In certain embodiments of all aspects of the present disclosure, the concentration ratio is calculated from at least two of the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:0), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0, Cer(d18:2 / 24:1). ), Cer(d18:2 / 24:0), Cer(d20:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6, PC16:0 / 16:0, PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:8.

[0140] In certain embodiments of all aspects of the present disclosure, the concentration ratio is calculated from two of the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and / or PC16:0 / 16:0. In certain embodiments of all aspects of the present disclosure, the concentration ratio is calculated from two of the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), PC16:0 / 22:5 and / or PC14:0 / 22:6. In certain embodiments of all aspects of the present disclosure, the concentration ratio may be, but is not limited to, Cer(d18:1 / 16:0) / PC16:0 / 22:5, Cer(d18:1 / 18:0) / PC14:0 / 22:6, and / or Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0). In certain embodiments of all aspects of the disclosure, the concentration ratios Cer(d18:1 / 16:0) / PC16:0 / 22:5, Cer(d18:1 / 18:0) / PC14:0 / 22:6 and Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0) are assayed. In certain embodiments of all aspects of the disclosure, the concentration ratios Cer(d18:1 / 16:0) / PC16:0 / 22:5, Cer(d18:1 / 18:0) / PC14:0 / 22:6 and Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0) are assayed and the concentration of PC16:0 / 16:0 is assayed.

[0141] In certain embodiments of all aspects of the present disclosure, the lipid combination is selected from any of the lipid combinations referenced in Tables 2-18. In certain embodiments of all aspects of the disclosure, the concentrations of at least two, at least three, at least four, at least five, or at least six of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC36:6, and PC38:5. In certain embodiments of all aspects of the disclosure, the concentrations of Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC36:6, and 38:5 are assayed. In certain embodiments of all aspects of the disclosure, the concentration ratios Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0), Cer(d18:1 / 16:0) / PC38:5 and Cer(d18:1 / 18:0) / PC36:6 are assayed, and the concentration of PC32:0 is assayed.

[0142] In certain embodiments of all aspects of the present disclosure, marker combinations can be constructed by linear combinations, regression models, other unsupervised or supervised frequentist or Bayesian statistical classification or machine learning methods, such as support vector machines, kernel estimation, decision trees, or neural networks. In certain embodiments of all aspects of the present disclosure, the combination of markers is a continuous or discrete scoring system based on Cer, PC, Cer / PC, Cer / Cer, PC / Cer, PC / PC or other cardiovascular biomarkers. For discrete scoring, points may be awarded based on intervals, quantiles or other cutoff values ​​with non-equal probabilities of the individual score components determined from the same or other populations. In certain embodiments of all aspects of the present disclosure, the combination of markers is scored in a scoring system (such as, but not limited to, points 0-12, 0-15, or 0-9) with points awarded based on the quartiles (Q1-Q4) of the overall study population. For example, if a person's lipid biomarkers fall in the highest quartile, the person receives 3 points. The same evaluation is performed for the other lipid biomarkers and summed.

[0143] In certain embodiments of all aspects of the disclosure, the score is based on at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten lipids selected from the following: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), PC16:0 / 22:5, PC14:0 / 22:6, Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 16:0, Cer(d18:1 / 22:0), Cer(d18:1 / 26:0), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0, Cer(d16:1 / 18:0, Cer(d1 6:1 / 24:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(d18:2 / 24:1), Ce r(d18:2 / 24:0), Cer(d20:1 / 24:1), PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3 , PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and PC40:8. In certain embodiments of all aspects of the present disclosure, the score is based on at least two, at least three, at least four, at least five, or at least six of the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, and / or PC14:0 / 22:6.

[0144] In certain embodiments of all aspects of the present disclosure, the score is based on the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5 and / or PC14:0 / 22:6. In certain embodiments of all aspects of the present disclosure, the score is based on the following lipid ratios: Cer(d18:1 / 16:0) / PC16:0 / 22:5, Cer(d18:1 / 18:0) / PC14:0 / 22:6 and / or Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0).

[0145] In certain embodiments of all aspects of the present disclosure, the score is based on at least two, at least three, at least four, at least five, at least six, or at least seven of the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6, and / or PC16:0 / 16:0. In certain embodiments of all aspects of the present disclosure, the score is based on the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and / or PC16:0 / 16:0. In certain embodiments of all aspects of the present disclosure, the score is based on the following lipid ratios and individual lipids: Cer(d18:1 / 16:0) / PC16:0 / 22:5, Cer(d18:1 / 18:0) / PC14:0 / 22:6, Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0) and / or PC16:0 / 16:0.

[0146] In certain embodiments of all aspects of the present disclosure, the score is based on the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC38:5 and / or PC36:6. In certain embodiments of all aspects of the present disclosure, the score is based on the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC38:5, PC36:6 and / or PC32:0. In certain embodiments of all aspects of the present disclosure, the score is based on the following lipid ratios: Cer(d18:1 / 16:0) / PC38:5, Cer(d18:1 / 18:0) / PC36:6 and / or Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0).

[0147] In certain embodiments of all aspects of the disclosure, the score is constructed according to Tables 8, 9, 10 or 11. In certain embodiments of all aspects of the present disclosure, the score further comprises determining other cardiovascular biomarkers, including, but not limited to, triglycerides (TG), C-reactive protein (CRP), troponin T (TNT or TnT), troponin I (TNI or TnI), B-type natriuretic peptide (BNP), N-terminal pro-B-type natriuretic peptide (NT-proBNP), cystatin C, glycated hemoglobin A1c (HbA1c), glucose, tumor suppressor 2 (St2), galectin, trimethylamine-N-oxide (TMAO), lipoprotein-associated phospholipase A2 (Lp-PLA2), growth differentiation factor 15 (GDF15), lipoprotein(a) (Lp(a)), any other lipoprotein subgroup composition or particle number, or any combination thereof.

[0148] In certain embodiments of all aspects of the present disclosure, the score may further include determining any other cardiovascular or metabolic biomarkers, including but not limited to creatine kinase (CK). In certain embodiments of all aspects of the present disclosure, the score further comprises TNT. In certain embodiments of all aspects of the present disclosure, the score further includes using other personal information or health data, such as gender, age, blood pressure, BMI, smoking status, diabetes, lipid-lowering treatments or other medications, history of cardiovascular events, ethnic minority background, geographic location, medical imaging data, such as angiographic or computed tomography (CT) medical imaging data, or any combination thereof. In certain embodiments of all aspects of the present disclosure, the score may further include any personal information or health data, including, but not limited to, medical history of CVD and / or CV events, family history of CVD and / or CV events, family history of diabetes, family history of other diseases. In certain embodiments of all aspects of the present disclosure, the score further includes medical image data. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0149] The present disclosure is based on the discovery that certain molecules are expressed at higher levels and certain molecules are expressed at lower levels in biological samples, for example, in the blood of subjects at risk of developing one or more future cardiovascular (CV) events, compared to controls. The combination derived from the concentrations of these molecules has surprisingly been found to be a good predictive biomarker of cardiovascular events, as described herein.

[0150] According to the present disclosure, the combination of at least one ceramide (Cer) and at least one phosphatidylcholine (PC) concentration provides a specific and sensitive diagnostic test that can be used to identify subjects at risk of developing a cardiovascular event. The biomarker combination of the present disclosure can be used to predict CV events when the levels of traditional markers, such as LDL-C (low density lipoprotein cholesterol), HDL-C (high density lipoprotein cholesterol) and total cholesterol, are normal, increased or decreased. The biomarker combination of the present invention can also be used for subjects with or without traditional risk factors, such as diabetes, hypertension or smoking. In addition, the predictive test according to the present disclosure can be advantageously used for patients undergoing statin treatment to assess whether they remain at risk of developing one or more cardiovascular events. The test can also be used for cardiovascular disease patients who have experienced a CV event to determine whether they have a high risk of developing one or more subsequent CV events.

[0151] A method for predicting one or more coronary artery disease events according to the present disclosure includes measuring the concentration of at least one ceramide and at least one phosphatidylcholine molecule, and optionally at least one additional ceramide and / or phosphatidylcholine, from a sample, e.g., a blood sample, e.g., a serum and / or plasma sample. A novel combination derived from the concentration of specific Cer and PC molecules is used as a marker for future cardiovascular events.

[0152] According to the method of the present disclosure, lipid biomarkers, such as lipid concentration, can be collected from a sample of interest through various chemical and high-resolution analytical techniques. Particularly suitable analytical techniques include, but are not limited to, mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy. In fact, any high-resolution technique that can resolve individual lipid species and provide their structural information can be used to determine lipid markers according to the present disclosure. For the purpose of the method of the present disclosure, lipid concentration is typically determined by using mass spectrometry. MS equipment can be coupled with direct sample injection, such as robotic nanoflow ion source equipment, or high-performance separation, such as LC, GC, HPLC, UHPLC, or UPLC. In addition, the use of nuclear magnetic resonance spectroscopy, fluorescence spectroscopy or dual polarization interferometry, high-performance separation, such as HPLC, UHPLC, or UPLC, immunoassays, such as ELISA, and / or binding moieties that can specifically bind to lipid analytes are useful in this context.

[0153] In the present disclosure, a novel combination of lipid biomarkers determined from biological samples, such as plasma or serum samples, is identified. Surprisingly, the combination of ceramide and phosphatidylcholine concentrations showed superior performance as a predictive biomarker for cardiovascular events compared to existing and conventionally used biomarkers.

[0154] Abbreviation As used herein, "ACS" is acute coronary syndrome, "AMI" is acute myocardial infarction, "ApoA" is apolipoprotein A, "ApoAI" is apolipoprotein AI, "ApoAII" is apolipoprotein AII, "ApoB" is apolipoprotein B, "ApoC" is apolipoprotein C, "BMI" is body mass index, "BNP" is B-type natriuretic peptide, and "ApoA" is apolipoprotein B. 13 "C" is carbon-13 isotope, "CAC" is coronary artery calcification, "CAD" is coronary artery disease, "Cer" is ceramide, "CETP" is cholesteryl ester transfer protein, "CHD" is coronary heart disease, "CRP" is C-reactive protein, "CT" is computed tomography, "CV" is cardiovascular, "CVD" is cardiovascular disease / coronary vascular disease, "ELISA" is enzyme-linked immunosorbent assay, "FA" is fatty acid, "GC" is gas chromatography, "GCxGC" is two-dimensional gas chromatography, "GDF15" is growth differentiation factor 15, and " 2 "H" is deuterium, "HbA1c" is glycated hemoglobin A1c, "HDL" is high density lipoprotein, "HDL-C" is high density lipoprotein cholesterol, "HDL-P" is high density lipoprotein particle or high density lipoprotein particle number, "HMG-CoA" is 3-hydroxy-3-methyl-glutaryl-coenzyme A, "HPLC" is high performance liquid chromatography, "HR" is hazard ratio, "IHD" is ischemic heart disease, and "IL-1mAb" is interleukin-1 monoclonal antibody. "LC" is liquid chromatography, "LCxLC" is two-dimensional liquid chromatography, "LDL" is low density lipoprotein, "LDL-C" is low density lipoprotein cholesterol, "LDL-P" is low density lipoprotein particle or low density lipoprotein particle number, "Lp(a)" is lipoprotein(a), "Lp-PLA2" is lipoprotein-associated phospholipase A2, "MI" is myocardial infarction, "MRM" is multiple reaction monitoring, "MS" is mass spectrometry, and " 15"N" is a nitrogen-15 isotope, "NMR" is nuclear magnetic resonance, "NT-proBNP" is N-terminal pro-B-type natriuretic peptide, "PC" is phosphatidylcholine, "PCSK9" is proprotein convertase subtilisin / kexin type 9, "RNA" is ribonucleic acid, "SB" is sphingoid base, "SCH-48461" is (3R,4S)-1,4-bis(4-methoxyphenyl)-3-(3-phenylpropyl)-2-azetidinone, "SD" is standard deviation, and "siRNA" is small interfering RNA (siRNA). NA, "St2" is tumor suppressor 2, "TC" is total cholesterol, "TG" is triglyceride, "TMAO" is trimethylamine-N-oxide, "TNF-alpha mAb" is tumor necrosis factor alpha monoclonal antibody, "TNI" or "TnI" is troponin I or high-sensitivity troponin I, "TNT" or "TnT" is troponin T or high-sensitivity troponin T, "UHPLC" is ultra-high performance liquid chromatography, and "UPLC" is ultra-high performance liquid chromatography.

[0155] Additionally, the following abbreviations are used in this disclosure: "ACE" is angiotensin-converting enzyme, "ACL" is adenosine triphosphate citrate lyase, "ALA" is alpha-linolenic acid, "ARB" is angiotensin II receptor blocker, "ATP" is adenosine triphosphate (adenocine), "CCTA" is computed tomography angiography of the coronary arteries, "CK" is creatine kinase, "DHA" is docosahexaenoic acid, and "E "CG or EKG" is electrocardiography, "EPA" is eicosapentaenoic acid, "GLP1" is glucagon-like peptide 1, "MRI" is magnetic resonance imaging, "NM" is nuclear medicine, "PET" is positron emission tomography, "SPECT" is single photon emission computed tomography, "SGLT2" is sodium-glucose transporter 2, "omega-3 or n-3" is omega-3, and "omega-6 or n-6" is omega-6.

[0156] definition In order that the present invention may be more readily understood, certain terms are defined herein. Additional definitions are set forth throughout the detailed description. As used herein, "cardiovascular disease (CVD)" has its common meaning in the art and is used to classify many conditions that affect the heart, heart valves, blood, and vascular system of the body, including coronary artery disease (CAD). CVD may or may not include atherosclerosis. In this disclosure, the terms CVD and CAD may be used interchangeably. As used herein, "coronary artery disease (CAD)", also known as "coronary heart disease (CHD)" and "ischemic heart disease (IHD)", is characterized by atherosclerosis in the coronary arteries. Atherosclerotic lesions narrow the coronary artery lumen, reducing blood flow to the heart. When the flow of oxygen-rich blood to the heart is reduced or blocked, angina or myocardial infarction can result. In this disclosure, "cardiovascular (CV) events" include stable angina, unstable angina, myocardial infarction (MI), acute myocardial infarction (AMI), acute coronary syndrome (ACS), stroke, transient ischemic attack, deep vein thrombosis, heart failure, hospitalization for heart failure, and cardiovascular death. Such events may include atherosclerosis. In this disclosure, the terms CV event, CVD event, CAD event, CHD event, CV complication, CVD complication, CAD complication, CHD complication, CV outcome, CVD outcome, CAD outcome, and CHD outcome may be used interchangeably. In this disclosure, "cardiovascular (CV) events" may also include peripheral arterial disease (also called peripheral arterial disease) and different forms of arrhythmias, such as atrial fibrillation.

[0157] As used herein, the terms "subject", "patient" and "individual" are used interchangeably herein to refer to any mammalian subject, particularly a human, for whom prediction, diagnosis or treatment is desired. The subject may be a healthy individual with no prior signs or symptoms of CVD. Or the subject may have previously suffered or is suffering from CVD or CAD. The subject may also be suspected of suffering from CVD and / or may be at high risk of developing a CV event. The subject may or may not have previously suffered from a cardiovascular event, such as angina, myocardial infarction or stroke. The subject may or may not suffer from a metabolic disease, such as diabetes. In addition, the subject may or may not have normal levels of traditional lipid markers, such as total cholesterol, LDL or HDL, and / or may or may not have traditional risk factors, such as diabetes, hypertension or smoking. The subject may or may not have received treatment, such as, but not limited to, statin treatment, or may be naive to any prior treatment or medication.

[0158] As used herein, a "sample" is a biological sample obtained from a subject or a group or population of subjects. The sample may be a blood sample, a serum sample, a plasma sample, a saliva sample, a urine sample, a tissue sample or a fraction thereof, such as a lipoprotein fraction. Serum and plasma samples are typical. The sample may be prepared using techniques well known in the art. In certain embodiments, the blood sample is a dried blood spot. In certain embodiments, the blood sample is dried on a filter. In some embodiments, the blood sample may be collected as dried plasma or serum on a card, which separates the plasma or serum from a drop of blood and dries the separated plasma or serum sample onto the card.

[0159] As used herein, a "control" may be a control sample. A control may also be a concentration determined from a sample from a single healthy individual. A control may also be a sample representing a combination of samples from a general population of healthy individuals. Alternatively, a control may be a control value, score, or set of data regarding a biomarker in a sample that has been previously determined, calculated, or extrapolated, or may not yet be determined, calculated, or extrapolated, or may be taken from the literature. In some embodiments, the control sample may be one previously obtained from the same subject, or the control value may be one previously taken from the same subject, while in other embodiments, the control sample or value may be one obtained from a different subject. "Control", i.e., control value or control sample, as used herein, is typically representative of a group of subjects or a population of subjects. In this context, "representative" means that the lipid concentration reflected by the control value compared in the context of the present disclosure corresponds to the average concentration value of the lipid concentration in the corresponding individual samples from the subjects of the group or population. Similarly, in the case of a control sample, "representative" means that the lipid concentration in the control sample compared in the context of the present disclosure corresponds to the average concentration of the lipid concentration in the corresponding individual samples from the subjects of the group or population. Typically, the concentration that is all lipid concentration in the control sample corresponds to the average concentration of the lipid concentration in the corresponding individual samples from the subjects of the group or population. An individual having such a value can be considered a "healthy individual" for the purposes of the present disclosure. A control sample is particularly suitable for comparison with a subject sample if it is obtained in the same or essentially the same manner from the same type of biological tissue or source, for example, if the subject sample is a serum or plasma sample, the corresponding control sample is also a serum or plasma sample, respectively.

[0160] In some embodiments, a control sample from a group of subjects or a control sample from a population of subjects in the sense of the present disclosure is obtained by mixing equal amounts of samples obtained directly from the subjects of said group or population, or by mixing equal amounts of fractions, constituents or reaction products thereof. It will be appreciated that a control value useful for the purposes of this disclosure is typically a value obtained or obtainable using any one of the appropriate control samples described herein. In the context of this disclosure, a control sample may be a sample from a healthy individual, a general population of healthy individuals, a CVD patient who has not yet experienced any major CV event, or a group of CVD patients who have not yet experienced any major CV event. In some embodiments of the methods described herein, the control may be a control value, e.g., a concentration score or combined value of at least one ceramide of formula I and at least one phosphatidylcholine of formula II, or a combined value of at least one ceramide of formula I and at least one phosphatidylcholine of formula II with at least one additional ceramide or phosphatidylcholine selected from any of the Cer and PC species mentioned in Table 1.

[0161] For the purposes of this disclosure, lipids are named according to the following nomenclature: Cer is ceramide and PC is phosphatidylcholine. As used herein, the nomenclature of ceramide (Cer) is presented as the number of the first pair corresponding to the sphingoid base (SB) and the number of the second pair corresponding to the fatty acid (FA) chain of the molecule. In the SB and FA nomenclature, the first number of each pair refers to the number of carbon atoms in the SB or FA chain, and the second number refers to the number of carbon-carbon double bonds in the SB or FA chain.

[0162] As used herein, ceramide (Cer) of Formula I refers to a ceramide species having the following structure: [ka] (In the formula, R 1is a saturated, monounsaturated or diunsaturated alkyl chain having 11 to 17 carbon atoms, R 2 is a saturated, monounsaturated or diunsaturated alkyl chain having 13 to 25 carbon atoms.

[0163] As used herein, the nomenclature of phosphatidylcholine (PC) is presented as either brutto species or molecular species. In brutto nomenclature, the first number refers to the total number of carbon atoms in the molecule, and the second number refers to the total number of carbon-carbon double bonds of the molecule. In molecular nomenclature, the first pair of numbers corresponds to the first fatty acid (FA) chain, and the second pair of numbers corresponds to the second FA chain of the molecule. In FA nomenclature, the first number of each pair refers to the number of carbon atoms in the FA chain, and the second number refers to the number of carbon-carbon double bonds of the FA chain. In molecular nomenclature, the first FA chain is represented by R in formula II. 1 or R 2 Thus, the second FA chain can also be R 1 or R 2 It may be either of the following. As used herein, phosphatidylcholine of formula II (PC) refers to a phosphatidylcholine species having the following structure: [ka] (In the formula, R 3 and R 4 is a saturated, monounsaturated or polyunsaturated alkyl chain.

[0164] "Treatment" and "treatment" are used interchangeably in this disclosure and may include any therapeutic treatment typically given to subjects at risk of developing or suffering from CVD, such as, but not limited to, administering a drug and / or providing a subject with treatment, behavioral and / or lifestyle modification. Behavioral and / or lifestyle modification may include, for example, but not limited to, lifestyle intervention and / or counseling, including instruction, encouragement and / or follow-up regarding healthy diet, weight management, physical activity / exercise and / or smoking cessation. Treatment may also be surgery, such as angioplasty, stent placement or diversion surgery. Treatment may also include controlling the progression of CVD and / or its complications. As used herein, treatment may also be any cardiac and / or vascular surgery, such as, but not limited to, revascularization, e.g., angioplasty, stent placement or bypass surgery, and any lifestyle modification, such as, but not limited to, lifestyle intervention and / or counseling including instruction, encouragement and / or follow-up regarding healthy diet, weight management, physical activity / exercise, stress management and / or smoking cessation.

[0165] Treatment may further include any cardiac and / or vascular imaging and testing, such as, but not limited to, ultrasound, echocardiography, angiography, electrocardiography (ECG or EKG), cardiac stress testing, computed tomography (CT), coronary CT calcium scan, coronary computed tomography angiography (CCTA), magnetic resonance imaging (MRI), nuclear medicine (NM) imaging, positron emission tomography (PET), single photon emission computed tomography (SPECT), and / or myocardial perfusion imaging. Treatment may also include providing any medical and / or lifestyle management services to the subject. Treatment may be administered by any health care provider, including, but not limited to, a clinician, physician, medical doctor, therapist, nurse, psychologist, physical therapist, personal trainer, and / or nutritionist. Treatment may also include preventative measures, such as, but not limited to, administering a drug and / or providing the subject with therapy, behavioral and / or lifestyle modifications. Behavioral and / or lifestyle modifications may include, for example, lifestyle interventions and / or counseling, including, but not limited to, instructions, encouragement and / or follow-up regarding healthy eating, weight management, physical activity / exercise, stress management and / or smoking cessation. Treatment or prevention can further include, for example, requesting testing by a commercial laboratory, providing an assay result useful for determining whether a subject is in need of treatment or prevention.

[0166] "Drug," "pharmaceutical product," "medicine," "medication," and "drug" are used interchangeably in this disclosure and may include any pharmaceutical agent typically administered to subjects with CVD, e.g., a statin, another lipid-lowering / modifying drug, and / or a modulator of lipid concentrations. As used herein, a drug may be, for example, an HMG-CoA reductase inhibitor (a statin or other HMG-CoA reductase inhibitor), niacin (nicotinic acid), a cholesterol absorption inhibitor (ezetimibe), a cholesteryl ester transfer protein (CETP) inhibitor, a bile acid sequestrant, a fibrate, a phytosterol, an anti-inflammatory agent (e.g., methotrexate, IL-1 mAb, TNF-alpha mAb), acetylsalicylic acid (aspirin) or a PCSK9 inhibitor, or a modulator of lipid levels selected from small molecules, antibodies, antisense RNA, small interfering RNA (siRNA), and natural or modified lipids, or any combination thereof.

[0167] As used herein, a drug may also be any pharmaceutical agent for treating or preventing CVD, diabetes, metabolic syndrome or other metabolic disorders, including, but not limited to, anticoagulant and antithrombotic drugs (e.g., rivaroxaban), diabetes drugs (e.g., insulin, SGLT2 (sodium-glucose transporter 2) inhibitors, metformin, GLP1 (glucagon-like peptide 1) agonists), blood pressure drugs (e.g., diuretics, beta-receptor blockers, angiotensin-converting enzyme (ACE) inhibitors, etc.). In some embodiments, the therapeutic agents include, but are not limited to, anti-inflammatory drugs, angiotensin II receptor blockers (ARBs), calcium channel blockers, alpha blockers, beta receptor blockers, renin inhibitors, heart failure medications, omega-3 (ω-3 or n-3) and omega-6 (ω-6 or n-6) polyunsaturated fatty acids (e.g., eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), alpha-linolenic acid (ALA)), ATP (adenosine triphosphate) citrate lyase (ACL) inhibitors (e.g., bempedoic acid), antisense oligonucleotides, or any combination thereof.

[0168] As used herein, "statins" are a class of lipid-lowering drugs (HMG-CoA reductase inhibitors). Statins are selected from the group consisting of, but not limited to, atorvastatin, cerivastatin, fluvastatin, fluvastatin XL, lovastatin, pitavastatin, pravastatin, rosuvastatin and simvastatin, or any combination thereof. As used herein, the statin may also be mevastatin.

[0169] For purposes of this disclosure, the "cholesterol absorption inhibitor" may be ezetimibe or SCH-48461, the cholesteryl ester transfer protein (CETP) inhibitor may be evacetrapib, anacetrapib or dalcetrapib, the bile acid sequestrant may be colesevelam, cholestyramine or colestipol, the fibrate may be fenofibrate, gemfibrozil, clofibrate or bezafibrate, and the PCSK9 inhibitor is selected from, but is not limited to, a PCSK9-specific antibody, an siRNA, and a peptidomimetic. According to the present disclosure, a "modulator" may be a small molecule (<1500 Daltons molecular weight, typically <800 Daltons molecular weight), an antibody, an antisense RNA, a small interfering RNA (siRNA), or a natural or modified lipid. As used herein, "effectiveness of treatment" and "therapeutic effectiveness" are intended to mean the ability of a treatment to achieve the therapeutic objective for which it is administered.

[0170] As used herein, "CVD marker combination" refers to a particular combination of lipid biomarkers used for the methods and uses of the present disclosure. The CVD marker combination comprises at least one ceramide of formula I and at least one phosphatidylcholine of formula II. In some embodiments, the CVD marker combination further comprises at least one additional ceramide and / or at least one additional phosphatidylcholine selected from any of the Cer and PC species mentioned in Table 1 of the present disclosure. As used herein, the combination can be, for example, a ratio, sum, difference, product, remainder, value, score, calculation, formula, equation, algorithm, or any combination thereof of at least one ceramide of formula I and at least one phosphatidylcholine of formula II. In some embodiments, the combination can be calculated from the concentration of at least one ceramide of formula I and at least one phosphatidylcholine of formula II. In some embodiments, the CVD marker combination can be the concentration ratio of at least one ceramide of formula I and at least one phosphatidylcholine of formula II. In some embodiments, the combination further comprises at least one additional ceramide and / or at least one additional phosphatidylcholine selected from any of the Cer and PC species mentioned in Table 1 of the present disclosure.

[0171] In some embodiments, the at least one additional Cer is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional Cers selected from any of the Cer species mentioned in Table 1. In some embodiments, the at least one additional Cer is a combination including, but not limited to, a Cer / Cer ratio of at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional Cers. The combination may also be another combination as described herein.

[0172] In some embodiments, the at least one additional PC is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional PCs selected from any of the PC species mentioned in Table 1. In some embodiments, the at least one additional PC is a combination including, but not limited to, a PC / PC ratio of at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 additional PCs. The combination may also be another combination as described herein.

[0173] In some embodiments, the CVD marker combination further comprises other cardiovascular biomarkers, including, but not limited to, LDL, LDL-C, LDL-P, HDL, HDL-C, HDL-P, any other lipoprotein subgroup composition or particle number, TC, TG, CRP, troponin T, troponin I, BNP, NT-proBNP, cystatin C, HbA1c, glucose, St2, galectin, TMAO, Lp-PLA2, GDF15, Lp(a), ApoB, ApoAI, ApoAII, ApoC, or any combination thereof. In some embodiments, the CVD marker combination further comprises other personal or health data, such as gender, age, blood pressure, smoking status, diabetes, lipid-lowering treatment or other medications, history of cardiovascular events, ethnic minority background, geographic location, medical imaging data, such as from angiogram or computed tomography (CT), or any combination thereof. In some embodiments, marker combinations can be constructed by linear combinations, regression models, other unsupervised or supervised frequentist or Bayesian statistical classification or machine learning methods, such as support vector machines, kernel estimation, decision trees, or neural networks. In some embodiments, the CVD marker combination may further include any other cardiovascular or metabolic biomarker, including but not limited to creatine kinase (CK). In some embodiments, the combination of markers is a continuous or discrete scoring system based on Cer, PC, Cer / PC, Cer / Cer, PC / Cer, PC / PC or other cardiovascular biomarkers. For discrete scoring, points can be awarded based on intervals, quantiles or other cutoff values ​​with non-equal probabilities of the individual score components determined from the same or other populations. In this disclosure, the terms CVD marker combinations, CVD biomarker combinations, CV marker combinations, CV biomarker combinations, marker combinations, biomarker combinations, lipid marker combinations and combinations can be used interchangeably.

[0174] As used herein, "standard combination" refers to a particular combination of lipid standards used for the methods and uses of the present disclosure. The standard combination comprises at least one ceramide of formula I and at least one phosphatidylcholine of formula II. In some embodiments, the standard combination further comprises at least one additional ceramide and / or phosphatidylcholine selected from any of the Cer and PC species mentioned in Table 1 of the present disclosure. In some embodiments, one or more components of the standard combination are isotopically labeled. In some embodiments, the isotope of the isotope-labeled component is deuterium ( 2 In some embodiments, the isotope of the isotopically labeled component is 13 C or 15 It's N.

[0175] As used herein, a "composition" or a "kit" may be any combination of any standard, control, reagent, solution or solvent used for the methods and uses of the present disclosure. The composition or kit may include at least one ceramide of formula I and at least one phosphatidylcholine of formula II. In some embodiments, the composition or kit further includes at least one additional ceramide and / or phosphatidylcholine selected from any of the Cer and PC species mentioned in Table 1 of the present disclosure. In some embodiments, one or more components of the composition or kit are isotopically labeled. In some embodiments, the isotope of the isotopically labeled component is deuterium. In some embodiments, the isotope of the isotopically labeled component is hydrogen. 13 C or 15 It's N. As used herein, a composition or kit may further comprise a container, instructions for use for a method or use disclosed herein, and / or other elements for carrying out a method or use disclosed in this disclosure. In some embodiments, the composition or kit comprises elements for collecting a blood sample, e.g., a dried blood spot on a filter. The composition or kit may be purchased from a commercial manufacturer or prepared in-house in a laboratory. The composition or kit may be a test kit for laboratory use or a home test kit (commercial test). In certain embodiments, the composition or kit is a combination of any standard, control, reagent, solution or solvent used to assay the concentration of at least one ceramide of formula I and at least one phosphatidylcholine of formula II, as well as any at least one additional ceramide and / or phosphatidylcholine selected from any of the Cer and PC species mentioned in Table 1 of the present disclosure. The composition or kit can be used for assays carried out using various chemical and high resolution analytical techniques, including but not limited to mass spectrometry (MS) and nuclear magnetic resonance (NMR). MS analysis can be coupled to another high performance separation method, such as gas chromatography (GC), two-dimensional gas chromatography (GCxGC), liquid chromatography (LC), two-dimensional liquid chromatography (LCxLC), high performance liquid chromatography (HPLC) or ultra-high performance liquid chromatography (UHPLC).

[0176] As used herein, a "reagent" may be any standard, control, substance, compound, solution, solvent, agent, component, preparation, or any combination thereof used for the disclosed methods and uses. A reagent may also be a combination or mixture of any standard, control, substance, compound, solution, solvent, agent, component, and preparation used in the disclosed methods and uses. In some embodiments, one or more of any of the components of the reagent is isotopically labeled. In some embodiments, the isotope of the isotopically labeled component is deuterium. In some embodiments, the isotope of the isotopically labeled component is 13 C or 15 It's N. In some embodiments, the reagent is used to assay the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample. In some embodiments, the reagent is further used to assay the concentration of at least one additional ceramide and / or phosphatidylcholine selected from any of the Cer and PC species mentioned in Table 1 of the present disclosure. In some embodiments, the reagents are used in the preparation of reagents, kits, or compositions to practice the methods and uses of the present disclosure.

[0177] For purposes of this disclosure, the terms "obtaining data," "collecting data," "obtaining information," and "collecting information" may be used interchangeably. As used herein, "determining" in the context of the biomarkers disclosed herein refers to quantitatively or relatively determining the amount of the biomarker in a sample. For quantitative determination, either the absolute amount or the exact amount of the biomarker in the sample is determined. The relative amount or level of the biomarker in the sample may alternatively be determined, e.g., the amount of the biomarker in the sample is determined to be increased or decreased relative to a control as described herein.

[0178] As used herein, "assaying," "detecting," "analyzing," or "generating quantitative data" refers to measuring the amount, content, abundance, level, or concentration of a biomarker of the present disclosure in a sample using a testing apparatus, instrument, or device, such as, but not limited to, mass spectrometry. As used herein, "biomarker," "marker," "CVD marker," "CVD biomarker," "CV marker," "CV biomarker," "lipid marker," and "lipid biomarker" can be used interchangeably.

[0179] As used herein, the terms "combination," "group," "set," "composition," and "mixture" can be used interchangeably. As used herein, and in accordance with Table 1, "direction of change" refers to a change in the amount, content, abundance, level or concentration of a biomarker in a sample from a subject when the subject is at risk of developing one or more cardiovascular events, compared to a control. When assessing the effectiveness of a CVD treatment in a subject, according to some embodiments of the present disclosure, the direction of change is opposite to that presented in Table 1. As used herein, "risk" refers to low, moderate or high risk of developing one or more cardiovascular events.

[0180] The terms "the disclosure, description or invention," "in accordance with the disclosure, description or invention," "according to the disclosure, description or invention," and "the present disclosure, description or invention" as used herein are intended to refer to all aspects and embodiments of the disclosure described and / or claimed herein. As used herein, the term "comprising" may be interpreted as encompassing both "including" and "consisting of." EXAMPLES

[0181] Example 1 material and method Description of study cohort and samples Plasma samples were obtained from 3700 adult subjects undergoing coronary angiography for suspected stable angina who were followed for a mean of 10 years for cardiovascular (CV) mortality or events. In addition to the first study cohort, samples from another study cohort, the Population Study Cohort, were also analyzed. 7,300 serum samples from adult subjects in this cohort were analyzed. The subjects were followed for an average of 16 years for several cardiovascular outcomes, including myocardial infarction, heart failure, and CV mortality.

[0182] Analysis method For lipid quantification, serum samples were extracted using a modified Folch lipid extraction method performed on a Hamilton Microlab Star robot as described in Jung HR et al, High throughput quantitative molecular lipidomics. Biochim Biophys Acta. 2011 Nov; 1811 (11): 925-34, the entirety of which is incorporated herein by reference. Samples were spiked with known amounts of non-endogenous synthetic internal standards. After lipid extraction, samples were reconstituted in chloroform:methanol (1:2, v / v) and synthetic external standards were added to the extracts after extraction. Extracts were stored at -20°C prior to MS analysis. Ceramides and phospholipids (e.g., phosphatidylcholine) were analyzed on a hybrid triple quadrupole / linear ion trap mass spectrometer (5500 QTRAP) equipped with an ultra-high performance liquid chromatography (UHPLC) system (Eksigent ultraLC100 system) using multiple reaction monitoring (MRM). Ceramides were analyzed based on the negative ion mode method as described by Sullards MC et al., Structure-specific, quantitative methods for analysis of sphingolipids by liquid chromatography-tandem mass spectrometry: "inside-out" sphingolipidomics. Methods Enzymol.2007;432:83-115. Phospholipids were analyzed based on a previously published method, Niemi et al., Ovarian tumours of different histologic type and clinical stage induce similar changes in lipid metabolism. British Journal of Cancer. 2018;119:847-854.

[0183] statistical analysis All statistical analyses were performed using R software. Cox proportional hazards models (constructed using the R package survival) were used to determine the association of variables or combinations of these with the outcomes of interest. Hazard ratios (HRs) from Cox regression models were expressed per standard deviation (SD). Binary logistic regression models were used to construct linear combinations of variables, and C statistics were calculated for Cox regression models using the Hmisc package.

[0184] Example 2 result Tables 2-12 present the results for the first study cohort, and Tables 13-18 present the results for the second study cohort. Table 2 shows that the ratio of ceramide to phosphatidylcholine or ceramide to ceramide predicts CV events more accurately than the corresponding ceramide or phosphatidylcholine alone. In this example, the Cox model was adjusted for age and stratified for sex. As shown in Table 2, lipid ratios have lower p-values ​​and higher C-statistics compared to the individual lipid components.

[0185] Table 2. p-values ​​for lipid ratios are lower than for the corresponding lipids alone. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9] [Table 2-10]

[0186] In the following examples (Tables 3-12), logistic regression was used to construct combinations of variables, and unadjusted Cox regression models were used to evaluate the predictive value of individual lipids or lipid ratios or combinations of these. Results are presented as hazard ratios and C statistics for both total CV events and CV deaths.

[0187] Table 3 provides an example where the combination of two ceramide / phosphatidylcholine ratios results in a lower p-value and a higher C-statistic compared to the individual ceramide / phosphatidylcholine ratios alone. Furthermore, additional ceramide / ceramide ratios and individual phosphatidylcholines increase the predictive value. In addition, high sensitivity troponin (TNT) also leads to more accurate prediction. [Table 3]

[0188] Table 4 provides examples where the combination of ceramide / phosphatidylcholine and ceramide / ceramide ratios results in lower p-values ​​and higher C-statistics compared to the individual lipid ratios alone. Furthermore, additional ceramide / phosphatidylcholine ratios and individual phosphatidylcholines increase the predictive value. [Table 4]

[0189] Table 5 provides examples where combinations of individual lipid concentrations of ceramide and phosphatidylcholine result in lower p-values ​​and higher C-statistics when more lipids are incorporated into the model. [Table 5]

[0190] Table 6 provides examples where the combination of ceramide / ceramide and ceramide / phosphatidylcholine ratios results in lower p-values ​​and higher C-statistics compared to the individual lipid ratios alone. Furthermore, additional ceramide / phosphatidylcholine ratios and individual phosphatidylcholines increase the predictive value. [Table 6]

[0191] Table 7 provides examples where combinations of ceramide and phosphatidylcholine concentrations result in lower p-values ​​and higher C-statistics compared to the individual lipids alone. Furthermore, additional ceramide / phosphatidylcholine ratios increase the predictive value. [Table 7]

[0192] Tables 8 and 9 present a simplified scoring system (points 0-12 and 0-9, respectively), where points are awarded based on the quartiles (Q1-Q4) of the overall study population. For example, if a person's Cer(d18:1 / 16:0) / PC16:0 / 22:5 ratio belongs to the highest quartile, he or she receives 3 points. The same evaluation is performed for the other three lipid biomarkers and summed. [Table 8] [Table 9]

[0193] Tables 10 and 11 provide additional examples where TNT, also a non-lipid cardiovascular biomarker, has been added to the scoring system, in this case with a scoring system scale of 0-15 and 0-12, respectively. [Table 10] [Table 11]

[0194] Table 12 provides examples of predictive values ​​of the scoring systems. In this example, CV score (3 components) refers to the scoring system presented in Table 9, CV score (4 components) refers to the scoring system presented in Table 8, and CV score (4 components + TNT) refers to the scoring system presented in Table 10. As shown in this example, the scores give lower p-values ​​and higher C-statistics than, for example, individual ceramide / phosphatidylcholine ratios. [Table 12]

[0195] Tables 13-18 show the results of the second study cohort. Table 13 provides examples of combinations of ceramide and phosphatidylcholine predicting acute coronary syndrome in population study cohort subjects. Table 13. Hazard ratios (HR) and p-values ​​for the combination of ceramide and phosphatidylcholine in subjects who developed acute coronary syndrome during follow-up. CV score (four components) refers to the scoring system presented in Table 8 and CV score (three components) refers to the scoring system presented in Table 9. 95%CI = 95% confidence interval. [Table 13-1] [Table 13-2] [Table 13-3]

[0196] Table 14 provides examples of combinations of ceramide and phosphatidylcholine predicting cardiovascular mortality in population study cohort subjects. Table 14. Hazard ratios (HR) and p-values ​​for the combination of ceramide and phosphatidylcholine in subjects who died for cardiovascular reasons during follow-up. CV score (four components) refers to the scoring system presented in Table 8 and CV score (three components) refers to the scoring system presented in Table 9. 95%CI = 95% confidence interval. [Table 14-1] [Table 14-2] [Table 14-3] [Table 14-4]

[0197] Table 15 provides examples of ceramide and phosphatidylcholine combinations that predict heart failure in population study cohort subjects. Table 15. Hazard ratios (HRs) and p-values ​​for the combination of ceramide and phosphatidylcholine in subjects who developed heart failure during follow-up. CV score (four components) refers to the scoring system presented in Table 8, and CV score (three components) refers to the scoring system presented in Table 9. 95%CI = 95% confidence interval. [Table 15-1] [Table 15-2] [Table 15-3]

[0198] Table 16 provides examples of combinations of ceramide and phosphatidylcholine that predict myocardial infarction in population study cohort subjects. Table 16. Hazard ratios (HRs) and p-values ​​for the combination of ceramide and phosphatidylcholine in subjects who developed a myocardial infarction during follow-up. CV score (four components) refers to the scoring system presented in Table 8 and CV score (three components) refers to the scoring system presented in Table 9. 95%CI = 95% confidence interval. [Table 16-1] [Table 16-2] [Table 16-3]

[0199] Table 17 provides examples of ceramide and phosphatidylcholine combinations predicting peripheral arterial disease in population study cohort subjects. Table 17. Hazard ratios (HRs) and p-values ​​for the combination of ceramide and phosphatidylcholine in subjects who developed peripheral arterial disease during follow-up. CV score (four components) refers to the scoring system presented in Table 8 and CV score (three components) refers to the scoring system presented in Table 9. 95%CI = 95% confidence interval. [Table 17-1] [Table 17-2]

[0200] Table 18 provides examples of combinations of ceramide and phosphatidylcholine that predict stroke in population study cohort subjects. Table 18. Hazard ratios (HR) and p-values ​​for the combination of ceramide and phosphatidylcholine in subjects who developed a stroke during follow-up. CV score (four components) refers to the scoring system presented in Table 8 and CV score (three components) refers to the scoring system presented in Table 9. 95%CI = 95% confidence interval. [Table 18-1] [Table 18-2]

[0201] Those skilled in the art will recognize, or be able to determine using no more than routine experimentation, many equivalents to the specific embodiments described herein, both in the examples and throughout the text of the specification, which equivalents are considered to be within the scope of this disclosure and are intended to be encompassed by the following claims or the above-recited items.

Claims

1. 1. A method for determining a subject's risk of developing one or more cardiovascular (CV) events, comprising: (a) assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I; 【Chemistry 1】 (In the formula, R 1 is a saturated, monounsaturated or diunsaturated alkyl chain having 11 to 17 carbon atoms; R 2 is a saturated, monounsaturated or diunsaturated alkyl chain having 13 to 25 carbon atoms; (b) assaying a sample from the subject to determine the concentration of at least one phosphatidylcholine (PC) of formula II; 【Chemistry 2】 (In the formula, R 3 and R 4 is a saturated, monounsaturated or polyunsaturated alkyl chain, and (c) determining that the subject is at risk for developing one or more CV events if the sample contains an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II compared to the control. The method includes:

2. at least one Cer of formula I is at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten Cers of formula I; and / or 2. The method of claim 1, wherein the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II.

3. (i) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (ii) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1; and (iii) determining that the subject is at risk for developing one or more CV events if the sample contains an increased or decreased concentration of at least one additional Cer and / or additional PC set forth in Table 1 compared to the control. The method of claim 1 or 2, further comprising:

4. After determining steps (c) and / or (iii), (d) administering a treatment to the subject determined to be at risk for developing one or more CV events. The method of any one of claims 1 to 3, further comprising:

5. A method of treating cardiovascular disease (CVD) or preventing one or more cardiovascular (CV) events in a subject, comprising administering a treatment to the subject if the subject is identified as being at risk of developing one or more CV events by the method of any one of claims 1 to 3.

6. A method for detecting at least two lipids in a sample from a subject, the method comprising detecting the concentration of at least one Cer of formula I and at least one PC of formula II.

7. 7. The method of claim 6, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

8. The method according to claim 6 or 7, further comprising detecting the concentration of at least one additional Cer and / or additional PC selected from any of the Cer and PC species mentioned in Table 1 in the sample.

9. 9. The method of any one of claims 6 to 8, comprising comparing the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten Cers of formula I, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten PCs of formula II, and / or the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 with a control.

10. The method according to any one of claims 6 to 9, further comprising using a standard combination comprising at least one Cer of formula I and / or at least one PC of formula II.

11. The method of claim 10, wherein the standard combination further comprises at least one additional Cer and / or PC selected from any of the Cer and PC species mentioned in Table 1.

12. (a) comparing the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten Cer of formula I, the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten PC of formula II, and / or the concentration of at least one additional Cer and / or PC selected from any of the Cer and PC species mentioned in Table 1 with a control; and (b) determining the subject as at risk for developing one or more CV events if the concentration of at least one Cer of formula I, at least one PC of formula II and / or at least one additional Cer and / or PC is altered compared to a control. The method of any one of claims 6 to 11, further comprising:

13. 1. A method of obtaining data for determining a subject's risk of developing one or more CV events, comprising: (a) assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I; (b) assaying a sample from the subject to determine the concentration of at least one phosphatidylcholine (PC) of formula II; and (c) determining that the subject is at risk for developing one or more CV events if the sample contains an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II compared to the control. The method includes:

14. 14. The method of claim 13, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

15. (a) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (b) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1; and (c) determining that the subject is at risk for developing one or more CV events if the sample contains an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to the control.

15. The method of claim 13 or 14, further comprising:

16. The method of any one of claims 13 to 15, further comprising obtaining a sample from a subject.

17. 1. A method for generating quantitative data regarding a subject, comprising: (a) assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I; and (b) assaying a sample from the subject to determine the concentration of at least one phosphatidylcholine (PC) of formula II. The method includes:

18. 18. The method of claim 17, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

19. (a) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (b) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1.

19. The method of claim 17 or 18, further comprising:

20. The method of any one of claims 17 to 19, wherein the assay is carried out by mass spectrometry (MS).

21. 1. A method for assessing the effectiveness of a CVD treatment in a subject, comprising: (a) assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I; (b) assaying a sample from the subject to determine the concentration of at least one phosphatidylcholine (PC) of formula II; and (c) determining that the treatment is effective if the sample contains a decreased concentration of at least one Cer of formula I and / or an increased concentration of at least one PC of formula II compared to the control. The method includes:

22. 22. The method of claim 21, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

23. (i) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (ii) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1; and (iii) determining that the treatment is effective if the sample contains an increased or decreased concentration of at least one additional Cer and / or at least one additional PC compared to the control.

23. The method of claim 21 or 22, further comprising:

24. The method of any one of claims 21 to 23, further comprising, after determining step (c) or (iii), (d) modifying, supplementing or maintaining a treatment already administered to the subject based on the concentration of the at least one Cer of formula I, the concentration of additional Cer, the concentration of the at least one PC of formula II and / or the concentration of additional PC obtained in said determining step (c) or (iii).

25. 24. The method of any one of claims 21 to 23, further comprising determining that the treatment is ineffective in the subject and increasing or changing the subject's treatment to a different treatment.

26. A method of treating CVD or preventing one or more CV events in a subject undergoing treatment for CVD, comprising administering a drug and / or providing a therapeutic, behavioral and / or lifestyle modification to the subject if the subject is identified as being ineffectively treated for CVD by the method of any one of claims 21 to 23.

27. 1. A method for selecting an appropriate CVD treatment for a subject, comprising: (a) assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I; (b) assaying a sample from the subject to determine the concentration of at least one phosphatidylcholine (PC) of formula II; and (c) determining that the subject requires treatment, or modification or supplementation of an already administered treatment, if the sample contains an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II compared to the control; The method includes:

28. 28. The method of claim 27, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

29. (i) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (ii) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1; (iii) determining that the subject requires treatment, or modification or supplementation of an already administered treatment, if the sample contains an increased or decreased concentration of at least one additional Cer and / or at least one additional PC as set forth in Table 1 compared to the control; 29. The method of claim 27 or 28, further comprising:

30. The method according to any one of claims 27 to 29, further comprising, after determining step (c) or (iii), a step of: (d) treating the subject based on the concentration of the at least one Cer of formula I, the concentration of the additional Cer, the concentration of the at least one PC of formula II and / or the concentration of the additional PC obtained in said determining step (c) or step (iii).

31. 30. The method of any one of claims 27 to 29, further comprising determining that the subject is in need of a treatment, or an alteration or supplementation of an already administered treatment, and administering to the subject the treatment determined to be in need thereof.

32. A method of treating CVD or preventing one or more CV events in a subject, comprising administering a drug to the subject and / or providing therapeutic, behavioral and / or lifestyle modifications if the subject is identified as needing a change in, or a supplement to, an already administered treatment according to the method of any one of claims 27 to 29.

33. 1. A method of collecting data for determining a subject's risk of developing one or more cardiovascular (CV) events, comprising: (a) assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I; (b) assaying a sample from the subject to determine the concentration of at least one phosphatidylcholine (PC) of formula II; and (c) determining that the subject is at risk for developing one or more CV events if the sample contains an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II compared to the control. The method includes:

34. at least one Cer of formula I is at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten Cers of formula I; and / or 34. The method of claim 33, wherein the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

35. (i) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (ii) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1; and (iii) determining that the subject is at risk for developing one or more CV events if the sample contains an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to the control.

35. The method of claim 33 or 34, further comprising:

36. A method of collecting data to detect the concentration of at least one Cer of formula I and at least one PC of formula II in a sample from a subject.

37. 37. The method of claim 36, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

38. 38. The method of claim 36 or 37, further comprising detecting the concentration of at least one additional Cer and / or PC selected from any of the Cer and PC species mentioned in Table 1 in the sample.

39. 1. A method of collecting data for assessing the effectiveness of a CVD treatment in a subject, comprising: (a) assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I; (b) assaying a sample from the subject to determine the concentration of at least one phosphatidylcholine (PC) of formula II; and (c) determining that the treatment is effective if the sample contains a decreased concentration of at least one Cer of formula I and / or an increased concentration of at least one PC of formula II compared to the control. The method includes:

40. at least one Cer of formula I is at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten Cers of formula I; and / or 40. The method of claim 39, wherein the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 PCs of formula II.

41. (i) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (ii) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1; and (iii) determining that the treatment is effective if the sample contains an increased or decreased concentration of at least one additional Cer and / or at least one additional PC compared to the control.

41. The method of claim 39 or 40, further comprising:

42. 1. A method of collecting data for selecting an appropriate CVD treatment for a subject, comprising: (a) assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I; (b) assaying a sample from the subject to determine the concentration of at least one phosphatidylcholine (PC) of formula II; and (c) determining that the subject requires treatment, or modification or supplementation of an already administered treatment, if the sample contains an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II compared to the control; The method includes:

43. 43. The method of claim 42, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula 2 is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

44. (i) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (ii) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1; and (iii) determining that the subject requires treatment, or modification or supplementation of an already administered treatment, if the sample contains an increased or decreased concentration of at least one additional Cer and / or at least one additional PC as set forth in Table 1 compared to the control; 44. The method of claim 42 or 43, further comprising:

45. A cardiovascular disease (CVD) marker combination for determining a subject's risk of developing one or more cardiovascular events, comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II.

46. 46. ​​The CVD marker combination of claim 45, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

47. 47. The CVD marker combination of claim 45 or 46, further comprising at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1.

48. 1. Use of a cardiovascular disease (CVD) marker combination for determining a subject's risk of developing one or more cardiovascular (CV) events, the CVD marker combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising: (a) assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I; (b) assaying a sample from the subject to determine the concentration of at least one phosphatidylcholine (PC) of formula II; and (c) determining that the subject is at risk for developing one or more CV events if the sample contains an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II compared to the control. Including, use.

49. 49. Use according to claim 48, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

50. The CVD marker combination further comprises at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, and the use comprises: (i) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (ii) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1; and (iii) determining that the subject is at risk for developing one or more CV events if the sample contains an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to the control.

50. The use according to claim 48 or 49, comprising:

51. Use of a cardiovascular disease (CVD) marker combination to detect the concentration of at least one Cer of formula I and at least one PC of formula II in a sample from a subject.

52. 52. Use according to claim 51 , wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

53. 53. The use according to claim 51 or 52, further comprising detecting the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 in the sample.

54. 54. Use according to any one of claims 51 to 53, comprising comparing the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten Cers of formula I, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten PCs of formula II and / or the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 with a control.

55. 1. Use of a cardiovascular disease (CVD) marker combination to obtain data for determining a subject's risk of developing one or more cardiovascular (CV) events, the combination of CVD markers comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising: (a) assaying a sample from a subject to determine the concentration of at least one Cer of formula I; (b) assaying a sample from the subject to determine the concentration of at least one PC of formula II; and (c) determining that the subject is at risk for developing one or more CV events if the sample contains an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II compared to the control. Including, use.

56. 56. Use according to claim 55, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

57. The CVD marker combination further comprises at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, and the use comprises: (i) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (ii) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1; and (iii) determining that the subject is at risk for developing one or more CV events if the sample contains an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to the control.

57. The use according to claim 55 or 56, comprising:

58. 1. Use of a cardiovascular disease (CVD) marker combination to assess the effectiveness of a CVD treatment in a subject, comprising: (a) assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I; (b) assaying a sample from the subject to determine the concentration of at least one phosphatidylcholine (PC) of formula II; and (c) determining that the treatment is effective if the sample contains a decreased concentration of at least one Cer of formula I and / or an increased concentration of at least one PC of formula II compared to the control. Use including.

59. at least one Cer of formula I is at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten Cers of formula I; and / or 59. The use according to claim 58, wherein the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

60. (i) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (ii) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1; and (iii) determining that the treatment is efficacious if the sample contains an increased or decreased concentration of at least one additional Cer and / or at least one additional PC compared to the control.

60. The use of claim 58 or 59, further comprising:

61. 1. Use of a cardiovascular disease (CVD) marker combination to select an appropriate CVD treatment for a subject, comprising: (a) assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I; (b) assaying a sample from the subject to determine the concentration of at least one phosphatidylcholine (PC) of formula II; and (c) determining that the subject requires treatment, or modification or supplementation of an already administered treatment, if the sample contains an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II compared to the control; Use including.

62. 62. Use according to claim 61, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

63. (i) assaying the sample to determine the concentration of at least one additional Cer selected from any of the Cer species mentioned in Table 1; and / or (ii) assaying the sample to determine the concentration of at least one additional PC selected from any of the PC species mentioned in Table 1; and (iii) determining that the subject requires treatment, or modification or supplementation of an already administered treatment, if the sample contains an increased or decreased concentration of at least one additional Cer and / or at least one additional PC as set forth in Table 1 compared to the control; 63. The use of claim 61 or 62, further comprising:

64. A standard combination for determining a subject's risk of developing one or more cardiovascular (CV) events, comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II.

65. 65. The standard combination of claim 64, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

66. 66. The standard combination of claim 64 or 65, further comprising at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

67. A standard combination for assaying the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample from a subject in the preparation of a reagent, kit or composition for determining the risk of a subject to develop one or more cardiovascular (CV) events, the standard combination comprising at least one Cer of formula I and at least one PC of formula II.

68. 68. The standard combination of claim 67, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

69. 69. The standard combination of claim 67 or 68, further comprising at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

70. A standard combination for assaying the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample from a subject in the preparation of a reagent, kit or composition for evaluating the effectiveness of a CVD treatment in a subject, the standard combination comprising at least one Cer of formula I and at least one PC of formula II.

71. 71. The standard combination of claim 70, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

72. 72. The standard combination of claim 70 or 71, further comprising at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

73. A standard combination for assaying the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample from a subject in the preparation of a reagent, kit or composition for selecting an appropriate cardiovascular disease (CVD) treatment for the subject, the standard combination comprising at least one Cer of formula I and at least one PC of formula II.

74. 74. The standard combination of claim 73, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

75. 75. The standard combination of claim 73 or 74, further comprising at least one additional Cer and / or at least one additional PC selected from any of the additional Cer and PC species mentioned in Table 1.

76. 76. The standard combination according to any one of claims 64 to 75, wherein at least one Cer of formula I is isotopically labeled Cer, at least one PC of formula II is isotopically labeled PC and / or at least one additional Cer and / or at least one additional PC species mentioned in Table 1 is isotopically labeled Cer and / or isotopically labeled PC.

77. The isotopes of the at least one isotopically labeled Cer of formula I, the at least one isotopically labeled PC of formula II and / or the at least one additional isotopically labeled Cer and / or the at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1 are deuterium, 13 C or 15 The standard combination of claim 76, wherein N is N.

78. 65. Use of the standard combination according to claim 64, comprising assaying a sample from a subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject is at risk of developing one or more cardiovascular (CV) events.

79. 79. Use of a standard combination according to claim 78, wherein at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

80. 80. The use of the standard combination of claim 78 or 79, wherein the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC shown in Table 1 compared to a control indicates that the subject is at risk of developing one or more cardiovascular (CV) events.

81. Use of a standard combination for detecting the concentration of at least one Cer of formula I and at least one PC of formula II in a sample from a subject, wherein the standard combination comprises at least one Cer of formula I and at least one PC of formula II.

82. 82. Use according to claim 81, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

83. The use of claim 81 or 82, further comprising detecting the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 in the sample, and the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

84. 1. Use of a standard combination for assessing the effectiveness of a cardiovascular disease (CVD) treatment in a subject, the standard combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising assaying a sample from the subject to determine the concentration of the at least one ceramide (Cer) of formula I and the concentration of the at least one phosphatidylcholine (PC) of formula II, wherein a decreased concentration of at least one Cer of formula I and / or an increased concentration of at least one PC of formula II in the sample compared to a control indicates the effectiveness of said treatment.

85. 85. Use according to claim 84, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

86. The use of claim 84 or 85, wherein the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1, and the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC compared to the control indicates the effectiveness of the treatment.

87. Use of a standard combination to select an appropriate CVD treatment for a subject, the standard combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising assaying a sample from the subject to determine the concentration of the at least one ceramide (Cer) of formula I and the concentration of the at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject requires treatment, or a modification or supplementation of an already administered treatment.

88. 88. Use according to claim 87, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

89. The use of claim 87 or 88, wherein the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1, and the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC shown in Table 1 compared to the control indicates that the subject requires treatment, or a change or supplementation of an already administered treatment.

90. Use of a standard combination to obtain data for determining a subject's risk of developing one or more cardiovascular (CV) events, the standard combination comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, the use comprising assaying a sample from the subject to determine the concentration of the at least one ceramide (Cer) of formula I and the concentration of the at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject has a risk of developing one or more cardiovascular events.

91. 91. Use according to claim 90, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

92. The use of claim 90 or 91, wherein the standard combination further comprises at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1, and the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC shown in Table 1 compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

93. 93. Use according to any one of claims 78 to 92, wherein at least one Cer of formula I is isotopically labelled Cer, at least one PC of formula II is isotopically labelled PC and / or at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 is isotopically labelled Cer and / or isotopically labelled PC.

94. At least one isotopically labeled isotope of Cer of formula I, at least one isotopically labeled isotope of PC of formula II and / or at least one additional isotopically labeled Cer and / or at least one additional isotopically labeled isotope of PC mentioned in Table 1 is deuterium, 13 C or 15 The use of claim 93, wherein said amino acid is N.

95. Use of one or more reagents in the preparation of a reagent, kit or composition for determining a subject's risk of developing one or more cardiovascular (CV) events, the use comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

96. 96. Use according to claim 95, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

97. The use of claim 95 or 96, wherein the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC set forth in Table 1 compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

98. Use of one or more reagents in the preparation of a reagent, kit or composition for detecting the concentration of at least one ceramide of formula I and at least one phosphatidylcholine of formula II in a sample from a subject.

99. 99. Use according to claim 98, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

100. 100. The use of claim 98 or 99, further comprising detecting the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1 in the sample.

101. Use according to any one of claims 98 to 100, comprising detecting an altered concentration of at least one Cer of formula I and / or an altered concentration of at least one PC of formula II in a sample from a subject.

102. Use according to any one of claims 98 to 101, comprising detecting an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in a sample from a subject.

103. Use according to any one of claims 98 to 102, comprising using a standard combination comprising at least one Cer of formula I and / or at least one PC of formula II.

104. Use of one or more reagents in the preparation of a reagent, kit or composition for obtaining data for determining a subject's risk of developing one or more cardiovascular (CV) events, the use comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

105. Use according to claim 104, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

106. The use of claim 104 or 105, wherein the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC shown in Table 1 compared to a control indicates that the subject is at risk of developing one or more cardiovascular events.

107. Use of one or more reagents in the preparation of a reagent, kit or composition for assessing the effectiveness of a cardiovascular disease (CVD) treatment in a subject, comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and the concentration of at least one phosphatidylcholine (PC) of formula II, wherein a decreased concentration of at least one Cer of formula I and / or an increased concentration of at least one PC of formula II in the sample compared to a control indicates the effectiveness of said treatment.

108. Use according to claim 107, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

109. The use of claim 107 or 108, wherein the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC compared to a control indicates the effectiveness of the treatment.

110. Use of one or more reagents in the preparation of a reagent, kit or composition for selecting an appropriate cardiovascular disease (CVD) treatment for a subject, the method comprising assaying a sample from the subject to determine the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II, wherein an increased concentration of at least one Cer of formula I and / or a decreased concentration of at least one PC of formula II in the sample compared to a control indicates that the subject requires treatment, or a modification or supplementation of an already administered treatment.

111. Use according to claim 110, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

112. The use of claim 110 or 111, wherein the use further comprises assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC biomarker selected from any of the Cer and PC species mentioned in Table 1, wherein an increased or decreased concentration of at least one additional Cer and / or at least one additional PC shown in Table 1 compared to a control indicates that the subject requires treatment, or a change or supplementation of an already administered treatment.

113. A composition or kit for diagnosing, predicting or detecting one or more cardiovascular (CV) events in a subject, or for carrying out any of the methods or uses according to any one of claims 1 to 112, comprising at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II.

114. The composition or kit of claim 113, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

115. 115. The composition or kit of claim 113 or 114, further comprising at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

116. 116. The composition or kit of any one of claims 113 to 115, wherein at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten Cer of formula I are isotopically labeled Cer and at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten PC of formula II are isotopically labeled PC and / or at least one additional Cer of formula I and / or at least one additional PC of formula II selected from any of the Cer and PC species mentioned in Table 1 is isotopically labeled Cer and / or isotopically labeled PC.

117. at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten isotopically labeled Cer of formula I, at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten isotopically labeled PC of formula II and / or at least one additional isotopically labeled Cer and / or at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1, wherein the isotope of 13 C or 15 The composition or kit of claim 116, wherein N is N.

118. A composition or kit according to any one of claims 113 to 117, further comprising standards, controls, reagents, solutions, solvents, containers, instructions for use and / or other elements for carrying out the method or use according to any one of claims 1 to 117.

119. A composition or kit according to any one of claims 113 to 118, comprising elements for collecting a blood sample.

120. 120. A composition or kit according to any one of claims 113 to 119, comprising an element for collecting dried blood spots on a filter.

121. 121. The composition or kit of any one of claims 113 to 120 which is a test kit for use in a laboratory or a home test kit.

122. A composition or kit according to any one of claims 113 to 121 which is any standard, control, reagent, solution or solvent combination used for diagnosing, predicting or detecting one or more cardiovascular (CV) events in a subject or for carrying out any of the methods or uses according to any one of claims 1 to 121.

123. 123. The composition or kit of any one of claims 113 to 122, which is any combination of standards, controls, reagents, solutions or solvents used to assay a sample to determine the concentration of at least one Cer of formula I and at least one PC of formula II.

124. 124. A composition or kit according to any one of claims 113 to 123, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

125. The composition or kit of claim 123 or 124, further used to assay the sample to determine the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

126. 126. The composition or kit of any one of claims 122 to 125, wherein any standard, control, reagent, solution or solvent combination is purchased from a commercial manufacturer or prepared in-house in-house laboratory.

127. 128. A composition or kit according to any one of claims 113 to 127 for use in a mass spectrometry (MS) or nuclear magnetic resonance (NMR) assay.

128. The composition or kit of claim 127, wherein mass spectrometry (MS) is coupled with gas chromatography (GC), two-dimensional gas chromatography (GCxGC), liquid chromatography (LC), two-dimensional liquid chromatography (LCxLC), high performance liquid chromatography (HPLC) or ultra-high performance liquid chromatography (UHPLC).

129. 33. The method of any one of claims 4, 5, 24-26 or 30-32, further comprising requesting testing by a laboratory that provides assay results useful for determining the concentration of at least one Cer of formula I and the concentration of at least one PC of formula II, and optionally administering a treatment to the subject if the subject has an increased concentration of at least one Cer of formula I and a decreased concentration of at least one PC of formula II compared to the control.

130. 130. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 129, wherein the cardiovascular (CV) events include stable angina, unstable angina, myocardial infarction (MI), acute myocardial infarction (AMI), acute coronary syndrome (ACS), stroke, transient ischemic attack, deep vein thrombosis, heart failure, hospitalization for heart failure and cardiovascular death.

131. 131. The method, use, CVD marker combination, standard combination, composition or kit of claim 130, wherein the cardiovascular (CV) event is induced by atherosclerosis.

132. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 131, wherein the subject is a healthy individual with no prior signs or symptoms of CVD and is suffering from, has previously suffered from, is suspected of suffering from, or has previously suffered from a CVD event.

133. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 132, wherein the subject suffers from diabetes.

134. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 133, wherein the subject is undergoing treatment.

135. 135. The method, use, CVD marker combination, standard combination, composition or kit of claim 134, wherein the subject is undergoing statin treatment.

136. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 135, wherein the subject has normal levels of traditional risk markers, such as total cholesterol, LDL and / or HDL, and / or is free of traditional risk factors, such as diabetes, hypertension or smoking.

137. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 136, wherein the sample is a biological sample.

138. 138. The method, use, CVD marker combination, standard combination, composition or kit of claim 137, wherein the sample is a blood sample, a serum sample, a plasma sample, a saliva sample, a urine sample, a tissue sample or a fraction thereof, such as a lipoprotein fraction.

139. 138. The method, use, CVD marker combination, standard combination, composition or kit of claim 137, wherein the sample is a dried blood spot.

140. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 139, wherein the control is a control sample or a control value, such as a concentration or score, obtained from an individual or a population of individuals.

141. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 140, wherein the control is obtained from a previously obtained sample from the same subject.

142. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 141, wherein the control is from a healthy individual, a general population of healthy individuals, a CVD patient who has not yet experienced any major CV event, or a group of CVD patients who have not yet experienced any major CV event.

143. 143. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 142, wherein treatment comprises administering a drug and / or providing a therapeutic, behavioral and / or lifestyle modification.

144. 144. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 143, wherein treatment comprises controlling the progression of CVD and / or its complications.

145. 144. The method, use, CVD marker combination, standard combination, composition or kit of claim 143, wherein the drug comprises an HMG-CoA reductase inhibitor (a statin or other HMG-CoA reductase inhibitor), niacin (nicotinic acid), a cholesterol absorption inhibitor (ezetimibe), a cholesteryl ester transfer protein (CETP) inhibitor, a bile acid sequestrant, a fibrate, a phytosterol, an anti-inflammatory agent (e.g. methotrexate, IL-1 mAb, TNF-alpha mAb), acetylsalicylic acid (aspirin) or a PCSK9 inhibitor, or a modulator of lipid levels selected from small molecules, antibodies, antisense RNA, small interfering RNA (siRNA), and natural or modified lipids, or any combination thereof.

146. 146. The method, use, CVD marker combination, standard combination, composition or kit of claim 145, wherein the statin comprises atorvastatin, cerivastatin, fluvastatin, fluvastatin XL, lovastatin, pitavastatin, pravastatin, rosuvastatin or simvastatin, or any combination thereof.

147. The method, use, CVD marker combination, standard combination, composition or kit of claim 145, wherein the cholesterol absorption inhibitor is ezetimibe or SCH-48461, the cholesteryl ester transfer protein (CETP) inhibitor is evacetrapib, anacetrapib or dalcetrapib, the bile acid sequestrant is colesevelam, cholestyramine or colestipol, the fibrate is fenofibrate, gemfibrozil, clofibrate or bezafibrate, and the PCSK9 inhibitor is a PCSK9 specific antibody, siRNA, or peptidomimetic.

148. 148. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 147, wherein the CVD marker combination comprises a ratio, sum, difference, product, remainder, value, score, calculation, formula, equation, algorithm, or any combination thereof of at least one Cer of formula I and at least one PC of formula II.

149. 149. The method, use, CVD marker combination, standard combination, composition or kit according to claim 148, wherein the CVD marker combination comprises a concentration ratio of at least one Cer of formula I and at least one PC of formula II.

150. The method, use, CVD marker combination, standard combination, composition or kit of claim 149, wherein the CVD marker combination further comprises at least one additional Cer and / or one additional PC selected from any of the Cer and PC species mentioned in Table 1.

151. 13. The use according to any one of claims 95 to 112, wherein the reagent comprises any standard, control, substance, compound, solution, solvent, agent, component, preparation, or any combination thereof, used for the methods and uses according to any one of claims 1 to 150.

152. 13. The use according to any one of claims 95 to 112, wherein the reagent comprises any standard, control, substance, compound, solution, solvent, agent, component or combination of preparations used for the methods and uses according to any one of claims 1 to 151.

153. Use according to any one of claims 95 to 112, wherein the reagent is used for assaying the concentration of at least one ceramide (Cer) of formula I and at least one phosphatidylcholine (PC) of formula II in a sample.

154. 154. The use of claim 153, further comprising assaying the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

155. 155. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 154, further comprising adding to the sample at least one isotopically labeled Cer of formula I and / or at least one isotopically labeled PC of formula II.

156. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 155, further comprising adding to the sample at least one additional isotopically labeled Cer and / or at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1.

157. The isotopes of the at least one isotopically labeled Cer of formula I, the at least one isotopically labeled PC of formula II, and / or the at least one additional isotopically labeled Cer and / or the at least one additional isotopically labeled PC are deuterium, 13 C or 15 The method or use of claim 155 or 156, wherein N is N.

158. 158. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 157, wherein the concentration of at least one Cer of formula I and the concentration of at least one PC of formula II are determined in one assay.

159. 159. The method, use, CVD marker combination, standard combination, composition or kit of claim 158, wherein one assay comprises determining the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten Cer of formula I and the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten PC of formula II.

160. The method, use, CVD marker combination, standard combination, composition or kit of claim 158 or 159, wherein one assay further comprises determining the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

161. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 158 to 160, wherein one assay is performed using mass spectrometry (MS).

162. 162. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 161, wherein the concentration is determined by using mass spectrometry, nuclear magnetic resonance spectroscopy, fluorescence spectroscopy or dual polarisation interferometry, high performance separation methods such as LC, GC, HPLC, UHPLC or UPLC, immunoassays such as ELISA and / or assays using binding moieties capable of specifically binding to the analyte.

163. (a) the method, use, CVD marker combination, standard combination, composition or kit further comprises determining the level of total cholesterol (TC), low density lipoprotein (LDL), low density lipoprotein cholesterol (LDL-C), high density lipoprotein (HDL), high density lipoprotein cholesterol (HDL-C), low density lipoprotein particle (LDL-P), high density lipoprotein particle (HDL-P), apolipoprotein B (ApoB), apolipoprotein AI (ApoAI), apolipoprotein AII (ApoAII), and / or apolipoprotein C (ApoC) in a sample from the subject; and / or (b) the subject does not have elevated levels of one or more of TC, LDL-C, ApoC or ApoB, or decreased levels of HDL-C; 163. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 162.

164. A method for monitoring the progression of a subject's risk of CVD and / or CV events, comprising assaying a sample from the subject to determine the concentration of at least one Cer of formula I and at least one PC of formula II, and comparing these concentrations with the concentrations in a sample previously obtained from the same subject.

165. The method of claim 164, wherein the at least one Cer of formula I is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 Cers of formula I and / or the at least one PC of formula II is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 PCs of formula II.

166. The method of claim 164 or 165, further comprising assaying the sample to determine the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

167. R of Formula II 3 and / or R 4 The method of claim 1, wherein is an alkyl chain containing 11 to 26 carbons.

168. R of Formula II 3 and / or R 4 The method of claim 1 , wherein is an alkyl chain containing up to six double bonds.

169. 169. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 168, wherein at least one Cer of formula I is Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(d18:2 / 24:1) and / or Cer(d20:1 / 24:1).

170. 170. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 169, wherein at least one PC of formula II is PC16:0 / 22:5, PC14:0 / 22:6, PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:

8.

171. 171. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 170, wherein at least one PC of formula II is PC16:0 / 22:5, PC14:0 / 22:6, PC16:0 / 22:6, PC16:0 / 20:4, PC18:0 / 20:4, PC17:0 / 20:3, PC36:8, PC36:6, PC36:4, PC38:4, PC38:7, PC34:4 and / or PC40:

8.

172. At least one Cer of formula I is Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(d18:2 / 24:1) and / or Cer(d20:1 / 24:1), and at least one PC of formula II is PC16:0 / 22:5, PC14:0 / 22:6, PC15:0 / 22:7, PC16:0 / 22:8, PC17:0 / 22:9, PC18:1 / 24:1, PC19:1 / 26:1, PC20:1 / 26:1, PC21:1 / 26:0, PC22:1 / 24:1, PC23:1 / 26:0, PC24:1 / 24:1, PC25:1 / 26:0, PC26:1 / 26:0, PC27:1 / 26:0, PC28:1 / 26:0, PC29:1 / 26:0, PC30:1 / 26:0, PC31:1 / 26:0, PC32:1 / 26:0, PC33:1 / 26:0, PC40:1 / 26:0, PC41:1 / 26:0, PC50:1 / 26:0, PC60:1 / 26:0, PC70:1 / 26:0, PC80:1 / 26:0, PC90:1 / 26:0, PC10:1 / 26:0, PC11 172. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 171, wherein the marker combination is C18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:

8.

173. At least one Cer of formula I is selected from the group consisting of Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(d18:2 / 24:1) and / or Cer(d20:1).

173. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 172, wherein the PC of formula II is PC16:0 / 22:5, PC14:0 / 22:6, PC16:0 / 22:6, PC16:0 / 20:4, PC18:0 / 20:4, PC17:0 / 20:3, PC36:8, PC36:6, PC36:4, PC38:4, PC38:7, PC34:4 and / or PC40:

8.

174. 174. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 173, wherein at least one Cer of formula I is Cer(d18:1 / 16:0), Cer(d18:1 / 18:0) and / or Cer(d18:1 / 24:1) and at least one PC of formula II is PC16:0 / 22:5 and / or PC14:0 / 22:

6.

175. At least one additional Cer is selected from the group consisting of Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 26:0), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:0), Cer(d1 175. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 174, wherein the markers are Cer(d18:2 / 16:0), Cer(d18:2 / 18:0), Cer(18:2 / 24:0), Cer(d18:2 / 24:1), Cer(d20:1 / 24:0) and / or Cer(d20:1 / 24:1).

176. 176. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 175, wherein the at least one additional PC is PC16:0 / 16:0, 16:0 / 22:5, PC14:0 / 22:6, PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:

8.

177. 177. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 176, wherein the concentrations of at least two, at least three, at least four, at least five, at least six or at least seven of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC14:0 / 22:6 and PC16:0 / 16:

0.

178. 178. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 177, wherein the concentrations of Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC14:0 / 22:6 and PC16:0 / 16:0 are assayed.

179. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 178, wherein the concentrations of at least two, at least three, at least four, at least five, at least six or at least seven of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC16:0 / 22:6 and PC16:0 / 16:

0.

180. 180. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 179, wherein the concentrations of Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC16:0 / 22:6 and PC16:0 / 16:0 are assayed.

181. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 180, wherein the concentrations of at least two, at least three, at least four, at least five, at least six or at least seven of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC16:0 / 22:6 and PC16:0 / 18:

3.

182. 182. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 181, wherein the concentrations of Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5, PC16:0 / 22:6 and PC16:0 / 18:3 are assayed.

183. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 182, wherein the concentrations of at least two, at least three, at least four, at least five or at least six of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), PC16:0 / 20:4, PC18:0 / 20:4 and PC38:

7.

184. 184. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 183, wherein the concentrations of Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), PC16:0 / 20:4, PC18:0 / 20:4 and PC38:7 are assayed.

185. 185. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 184, wherein the concentration ratio is calculated from the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten Cer of formula I, and / or the concentration of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine or at least ten PC of formula II.

186. 186. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 185, wherein the concentration ratio is calculated from the concentration of at least one additional Cer and / or at least one additional PC selected from any of the Cer and PC species mentioned in Table 1.

187. 187. The method, use, CVD marker combination, standard combination, composition or kit of claim 186, wherein the concentration ratio is Cer / Cer, Cer / PC, PC / Cer and / or PC / PC ratio.

188. 188. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 185 to 187, wherein the concentration ratio is selected from any of the lipid ratios mentioned in Tables 2, 3, 4, 6, 7, 8, 9, 10, 11 or 12.

189. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 185 to 188, wherein the concentration ratio is calculated from at least two of the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:0), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18:2 / 18:0 ... r(d18:2 / 24:1), Cer(d16:1 / 24:0) and / or Cer(d20:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6, PC16:0 / 16:0, PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22: 6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and / or PC40:

8.

190. 190. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 185 to 189, wherein the concentration ratio is calculated from two of the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and / or PC16:0 / 16:

0.

191. 191. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 185 to 190, wherein the concentration ratios are Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0), Cer(d18:1 / 16:0) / PC16:0 / 22:5 and / or Cer(d18:1 / 18:0) / PC14:0 / 22:

6.

192. 192. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 185 to 191, wherein the concentration ratios Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0), Cer(d18:1 / 16:0) / PC16:0 / 22:5 and Cer(d18:1 / 18:0) / PC14:0 / 22:6 are assayed and the concentration of PC16:0 / 16:0 is assayed.

193. 200. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 192, further comprising determining other cardiovascular biomarkers, such as triglycerides (TG), C-reactive protein (CRP), troponin T (TnT), troponin I (TnI), B-type natriuretic peptide (BNP), N-terminal pro-B-type natriuretic peptide (NT-proBNP), cystatin C, glycated hemoglobin A1c (HbA1c), glucose, tumor suppressor 2 (St2), galectin, trimethylamine-N-oxide (TMAO), lipoprotein-associated phospholipase A2 (Lp-PLA2), growth differentiation factor 15 (GDF15), lipoprotein(a) (Lp(a)), any other lipoprotein subgroup composition or particle number, or any combination thereof.

194. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 193, further comprising using other personal information or health data, such as gender, age, blood pressure, BMI, smoking status, diabetes, lipid lowering treatment or other medications, history of cardiovascular events, ethnic minority background, geographic location, medical imaging data, such as data from angiograms or computed tomography (CT), or any combination thereof.

195. 195. The method or use of any one of claims 1 to 194, wherein the treatment comprises surgery, such as angioplasty, stent placement or diversion surgery.

196. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 195, wherein the marker combination is constructed by linear combination, regression model, other unsupervised or supervised frequentist or Bayesian statistical classification or machine learning techniques, such as support vector machines, kernel estimation, decision trees or neural networks.

197. 200. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 196, wherein the marker combination is a continuous or discrete scoring system based on Cer, PC, Cer / PC, Cer / Cer, PC / Cer, PC / PC and / or other cardiovascular biomarkers.

198. The method, use, CVD marker combination, standard combination, composition or kit of claim 197, wherein the discrete scoring system is based on points being assigned based on intervals, quantiles or other cut-off values ​​having non-identical probabilities of the individual score components determined from the same or other populations.

199. 200. The method, use, CVD marker combination, standard combination, composition or kit of claim 197 or 198, wherein the marker combination is a scoring system in which points are assigned based on quartiles (Q1-Q4) of the overall study population.

200. 200. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 199, wherein the score is based on at least two, at least three, at least four, at least five, at least six or at least seven of the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and / or PC16:0 / 16:

0.

201. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 200, wherein the score is based on the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and / or PC16:0 / 16:

0.

202. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 201, wherein the score is based on the following lipid ratios and individual lipids: Cer(d18:1 / 16:0) / PC16:0 / 22:5, Cer(d18:1 / 18:0) / PC14:0 / 22:6, Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0) and / or PC16:0 / 16:

0.

203. 203. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 202, wherein the score is constructed according to Table 8, 9, 10 or 11.

204. 204. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 203, wherein the score further comprises determining other cardiovascular biomarkers, such as triglycerides (TG), C-reactive protein (CRP), troponin T (TNT or TnT), troponin I (TNI or TnI), B-type natriuretic peptide (BNP), N-terminal pro-B-type natriuretic peptide (NT-proBNP), cystatin C, glycated hemoglobin A1c (HbA1c), glucose, tumor suppressor 2 (St2), galectin, trimethylamine-N-oxide (TMAO), lipoprotein-associated phospholipase A2 (Lp-PLA2), growth differentiation factor 15 (GDF15), lipoprotein(a) (Lp(a)), any other lipoprotein subgroup composition or particle number, or any combination thereof.

205. 205. The method, use, CVD marker combination, standard combination, composition or kit of claim 204, wherein the score further comprises TNT.

206. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 205, wherein the score further comprises using other personal information or health data, such as gender, age, blood pressure, BMI, smoking status, diabetes, lipid lowering treatment or other medications, history of cardiovascular events, ethnic minority background, geographic location, medical imaging data, such as angiographic or computed tomography (CT) medical imaging data, or any combination thereof.

207. 207. The method, use, CVD marker combination, standard combination, composition or kit of claim 206, wherein the score further comprises medical image data.

208. Use according to any one of claims 95 to 112, wherein at least one Cer of formula I is isotopically labelled Cer, at least one PC of formula II is isotopically labelled PC and / or at least one additional Cer and / or at least one additional PC species mentioned in Table 1 is isotopically labelled Cer and / or isotopically labelled PC.

209. The isotopes of the at least one isotopically labeled Cer of formula I, the at least one isotopically labeled PC of formula II and / or the at least one additional isotopically labeled Cer and / or the at least one additional isotopically labeled PC selected from any of the Cer and PC species mentioned in Table 1 are deuterium, 13 C or 15 The use according to any one of claims 95 to 112, wherein N is N.

210. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 209, wherein the isotopically labeled Cer and / or PC is selected from the following isotopically labeled lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), Cer(d18:1 / 26:0), Cer(d18:1 / 26:1), Cer(d16:1 / 16:0), Cer(d16:1 / 18:0), Cer(d16:1 / 24:0), Cer(d16:1 / 24:1), Cer(d18:2 / 16:0), Cer(d18 :2 / 18:0), Cer(d18:2 / 24:1), Cer(d18:2 / 24:0), Cer(d20:1 / 24:1), PC16:0 / 22:5, PC14 :0 / 22:6, PC16:0 / 16:0, PC18:0 / 20:5, PC16:0 / 20:4, PC18:0 / 20:4, PC18:0 / 20:3, PC16:0 / 22:6, PC16:1 / 18:2, PC16:0 / 18:3, PC17:0 / 20:3, PC17:0 / 20:4, PC38:5, PC36:6, PC38:5, PC36:4, PC38:4, PC38:3, PC38:6, PC38:7, PC34:3, PC37:3, PC37:4, PC34:4 and PC40:

8.

211. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 210, wherein the isotopically labeled Cer and / or PC is selected from the following isotopically labeled lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and PC16:0 / 16:

0.

212. 212. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 211, wherein the isotopically labelled Cer and PC are Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and PC16:0 / 16:

0.

213. 213. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 212, wherein the isotopically labeled Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5, PC14:0 / 22:6 and / or PC16:0 / 16:0 is deuterium labeled.

214. 214. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 213, wherein the isotopically labelled Cer and / or PC is d7-Cer(d18:1 / 16:0), d7-Cer(d18:1 / 18:0), d7-Cer(d18:1 / 24:0), d7-Cer(d18:1 / 24:1), d9-PC16:0 / 22:5, d9-PC14:0 / 22:6 and / or d9-PC16:0 / 16:

0.

215. R of Formula I 1 is a saturated, monounsaturated or diunsaturated alkyl chain having 11 to 17 carbon atoms; R 2 is a saturated alkyl chain having from 13 to 21 carbon atoms, a monounsaturated or diunsaturated alkyl chain having from 13 to 25 carbon atoms.

216. R of Formula I 1 is a monounsaturated or diunsaturated alkyl chain having 13 to 15 carbon atoms, R 2 The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 215, wherein is a saturated, monounsaturated or diunsaturated alkyl chain having from 15 to 25 carbon atoms.

217. R of Formula I 1 is a monounsaturated or diunsaturated alkyl chain having 13 to 15 carbon atoms, R 2 is a saturated alkyl chain having from 15 to 21 carbon atoms, a monounsaturated or diunsaturated alkyl chain having from 15 to 25 carbon atoms.

218. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 217, wherein the concentrations of at least two, at least three, at least four, at least five or at least six of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5 and PC14:0 / 22:

6.

219. 219. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 218, wherein the concentrations of Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC16:0 / 22:5 and PC14:0 / 22:6 are assayed.

220. 192. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 185 to 191, wherein the concentration ratios Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0), Cer(d18:1 / 16:0) / PC16:0 / 22:5 and Cer(d18:1 / 18:0) / PC14:0 / 22:6 are assayed.

221. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 220, wherein the score is based on the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC16:0 / 22:5 and / or PC14:0 / 22:

6.

222. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 221, wherein the score is based on the following lipid ratios: Cer(d18:1 / 16:0) / PC16:0 / 22:5, Cer(d18:1 / 18:0) / PC14:0 / 22:6 and / or Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0).

223. 223. The method, use, CVD marker combination, standard combination, composition or kit according to any one of claims 1 to 222, wherein the lipid combination is selected from any of the lipid combinations mentioned in Tables 2 to 18.

224. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 223, wherein the concentrations of at least two, at least three, at least four, at least five or at least six of the following lipids are assayed: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC36:6 and PC38:

5.

225. 225. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 224, wherein the concentrations of Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1), Cer(d18:1 / 24:0), PC36:6 and 38:5 are assayed.

226. 226. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 225, wherein the concentration ratios Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0), Cer(d18:1 / 16:0) / PC38:5 and Cer(d18:1 / 18:0) / PC36:6 are assayed.

227. 227. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 226, wherein the concentration ratios Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0), Cer(d18:1 / 16:0) / PC38:5 and Cer(d18:1 / 18:0) / PC36:6 are assayed and the concentration of PC32:0 is assayed.

228. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 227, wherein the score is based on the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC38:5 and / or PC36:

6.

229. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 228, wherein the score is based on the following lipids: Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1), PC38:5, PC36:6 and / or PC32:

0.

230. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 229, wherein the score is based on the following lipid ratios: Cer(d18:1 / 16:0) / PC38:5, Cer(d18:1 / 18:0) / PC36:6 and / or Cer(d18:1 / 24:1) / Cer(d18:1 / 24:0).

231. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 230, wherein the PC may be a brutto PC or a molecular PC.

232. 232. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 231, wherein the cardiovascular (CV) event comprises stable angina, unstable angina, myocardial infarction (MI), acute myocardial infarction (AMI), acute coronary syndrome (ACS), stroke, transient ischemic attack, deep vein thrombosis, heart failure, hospitalization for heart failure, cardiovascular death, peripheral artery disease and / or arrhythmia.

233. 233. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 232, wherein the sample is dried plasma or serum collected on a card.

234. 33. The method of any one of claims 4, 5, 24-26 or 30-32, wherein treatment comprises providing any medical and / or lifestyle management services to the subject.

235. 144. The method, use, CVD marker combination, standard combination, composition or kit of claim 143, wherein the drug comprises any pharmaceutical agent for treating or preventing CVD, diabetes, metabolic syndrome or other metabolic disorder.

236. 236. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 235, further comprising determining other cardiovascular biomarkers, such as triglycerides (TG), C-reactive protein (CRP), troponin T (TnT), troponin I (TnI), B-type natriuretic peptide (BNP), N-terminal pro-B-type natriuretic peptide (NT-proBNP), cystatin C, glycated hemoglobin A1c (HbA1c), glucose, tumor suppressor 2 (St2), galectin, trimethylamine-N-oxide (TMAO), lipoprotein-associated phospholipase A2 (Lp-PLA2), growth differentiation factor 15 (GDF15), lipoprotein(a) (Lp(a)), any other lipoprotein subgroup composition or particle number, creatine kinase (CK), or any combination thereof.

237. The method, use, CVD marker combination, standard combination, composition or kit of any one of claims 1 to 236, further comprising using other personal information or health data, such as gender, age, blood pressure, BMI, smoking status, diabetes, lipid lowering treatment or other medications, medical history of CVD and / or CV events, ethnic minority background, geographic location, family history of CVD, CV events and / or diabetes, medical imaging data, such as data from angiograms or computed tomography (CT), or any combination thereof.

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