Application of biomarker cyclic dinucleotide cGAMP in diagnosis and risk prediction of myocardial infarction

Abstract

The invention provides application of a biomarker cyclic dinucleotide cGAMP in diagnosis and risk prediction of myocardial infarction. The application specifically relates to application of the biomarker cyclic dinucleotide cGAMP in preparation of a myocardial infarction diagnosis tool, application of the biomarker cyclic dinucleotide cGAMP in preparation of a myocardial infarction risk prediction product and application of the biomarker cyclic dinucleotide cGAMP in construction of a myocardial infarction risk prediction model. Experimental results show that the concentration of cGAMP in plasma of a patient suffering from coronary heart disease and the risk of suffering from myocardial infarction are in a remarkable U-shaped relationship, the risk of myocardial infarction can be increased compared with the cGAMP with too high or too low critical concentration, and the critical concentration is 1352 ng / L. Whether the plasma concentration is greater than or less than the critical concentration, the cGAMP has medium diagnostic ability to myocardial infarction.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the application of the biomarker cyclic dinucleotide cGAMP in the diagnosis and risk prediction of myocardial infarction. Background Art

[0002] Myocardial infarction (MI) is a myocardial cell necrosis caused by persistent myocardial ischemia and hypoxia due to acute occlusion or severe stenosis of the coronary artery. It is one of the most serious clinical types of coronary heart disease. According to the pathological mechanism and electrocardiographic manifestations, myocardial infarction can be divided into ST-segment elevation myocardial infarction (STEMI) and non-ST-segment elevation myocardial infarction (NSTEMI). Typical symptoms include severe chest pain, chest tightness, radiating pain (such as left shoulder, mandible or back), and accompanying dyspnea, nausea, cold sweat, etc., but its clinical manifestations are significantly heterogeneous. Some patients (especially the elderly, diabetic patients or women) may only show atypical symptoms such as fatigue, syncope or upper abdominal discomfort, or even be completely asymptomatic (i.e., "silent myocardial infarction"), making early identification difficult. In addition, chest pain symptoms are easily confused with gastroesophageal reflux, pulmonary embolism or musculoskeletal diseases, further increasing the risk of misdiagnosis.

[0003] At present, the diagnosis of myocardial infarction mainly relies on the combined analysis of clinical manifestations, dynamic changes of electrocardiograms, and myocardial injury markers (such as troponin and creatine kinase isoenzymes). Although electrocardiogram is the preferred tool for rapid screening in the emergency department, it may lack specific changes in the hyperacute phase, and individual anatomical variations (such as left bundle branch block) may interfere with interpretation. Although myocardial injury markers (such as high-sensitivity troponin) have significantly improved the diagnostic sensitivity, their levels usually increase significantly only a few hours after the onset of symptoms, which may lead to delays in early diagnosis. Imaging techniques (such as echocardiography, coronary artery CTA or angiography) can assist in assessing the extent of myocardial ischemia and vascular lesions, but they have limitations such as radiation exposure, invasive procedures (such as angiography), or equipment dependence, making them difficult to be widely used for rapid screening in pre-hospital or resource-poor areas.

[0004] Traditional risk assessment models (such as GRACE score and TIMI score) are mainly based on age, medical history, electrocardiogram and laboratory indicators, but their predictive effectiveness is limited by the insufficient coverage of new biomarkers (such as inflammatory factors, genetic markers) and individualized pathological mechanisms. In addition, people at high risk of myocardial infarction (such as patients with chronic kidney disease and diabetes) often have multi-system diseases, which further interferes with the accuracy of risk assessment. Since myocardial infarction is closely related to fatal complications such as malignant arrhythmias and cardiogenic shock, and the time window for reperfusion therapy directly affects the prognosis of patients, early accurate identification and risk stratification are of great significance to improving clinical outcomes and reducing medical costs. Therefore, exploring non-invasive and efficient new biomarkers or artificial intelligence-assisted diagnosis models has become a research hotspot in the current cardiovascular field. Summary of the invention

[0005] The purpose of the present invention is to provide a new biomarker cyclic dinucleotide cGAMP for risk prediction and disease diagnosis of myocardial infarction in patients with coronary heart disease, and to provide a new approach for diagnosis and risk prediction of myocardial infarction.

[0006] In order to achieve the above object, the present invention provides the use of a reagent for detecting the biomarker cyclic dinucleotide cGAMP in the preparation of a diagnostic tool for myocardial infarction.

[0007] Preferably, the reagent is used to detect the concentration of cyclic dinucleotide cGAMP in the plasma of patients with coronary heart disease.

[0008] Preferably, the reagent is an ELISA kit.

[0009] More preferably, the reagent uses a double antibody sandwich method to detect the concentration of cGAMP in the plasma of patients with coronary heart disease.

[0010] The double antibody sandwich method is as follows: the microplate is coated with purified cGAMP antibody to make a solid phase antibody, cGAMP is added to the microwells coated with the monoclonal antibody in sequence, and then combined with the HRP-labeled cGAMP antibody to form an antibody-antigen-enzyme-labeled antibody complex. After thorough washing, the substrate TMB is added for color development. TMB is converted into blue under the catalysis of HRP enzyme, and converted into the final yellow under the action of acid. The depth of color is positively correlated with the cGAMP in the sample. The absorbance is measured at a wavelength of 450nm using an enzyme reader, and the cGAMP concentration in the sample is calculated using the standard curve.

[0011] The present invention also provides the use of the biomarker cyclic dinucleotide cGAMP in the preparation of a myocardial infarction risk prediction product.

[0012] Preferably, the risk of suffering from myocardial infarction is predicted by detecting the concentration of cyclic dinucleotide cGAMP in the plasma of patients with coronary heart disease.

[0013] Preferably, the product for detecting the concentration of cyclic dinucleotide cGAMP in the plasma of patients with coronary heart disease is an ELISA kit.

[0014] Preferably, the myocardial infarction includes ST-segment elevation myocardial infarction and non-ST-segment elevation myocardial infarction.

[0015] Preferably, the concentration of the cyclic dinucleotide cGAMP in the plasma of patients with coronary heart disease has a significant U-shaped relationship with the risk of myocardial infarction. A cGAMP concentration that is too high or too low relative to the critical concentration will increase the risk of myocardial infarction. The critical concentration is 1352 ng / L.

[0016] Preferably, when the concentration of cyclic dinucleotide cGAMP = 1352 ng / L, the risk of myocardial infarction in patients with coronary heart disease is predicted to be low; when the concentration of cyclic dinucleotide cGAMP is <1352 ng / L, the concentration of cyclic dinucleotide cGAMP is inversely correlated with the risk of myocardial infarction; when the concentration of cyclic dinucleotide cGAMP is >1352 ng / L, the concentration of cyclic dinucleotide cGAMP is positively correlated with the risk of myocardial infarction.

[0017] The present invention also provides the use of the biomarker cyclic dinucleotide cGAMP in constructing a myocardial infarction risk prediction model.

[0018] Preferably, the input variables of the risk prediction model include plasma concentration values ​​of the cyclic dinucleotide cGAMP.

[0019] Preferably, the input variables of the risk prediction model also include age, gender, hypertension, diabetes, smoking, drinking, body mass index, systolic blood pressure, left ventricular ejection fraction, serum creatinine, cardiac troponin T, N-terminal pro-brain natriuretic peptide, high-density lipoprotein, apolipoprotein A1 and free fatty acids.

[0020] In the present invention, the biomarker cyclic dinucleotide cGAMP for diagnosing myocardial infarction and predicting the risk of myocardial infarction includes the following steps: step 1), using an anticoagulation tube to collect venous blood from fasting patients; step 2), separating plasma from the venous blood and performing cGAMP quantitative detection; step 3), determining the risk of myocardial infarction in patients with coronary heart disease based on the cGAMP quantitative detection results combined with a U-shaped curve, wherein the U-shaped curve is a curve describing the concentration of the cyclic dinucleotide cGAMP and the risk of myocardial infarction.

[0021] The beneficial effects of the present invention include at least:

[0022] 1. The present invention discovers for the first time that the concentration of cGAMP in the plasma of patients with coronary heart disease has a significant U-shaped relationship with the risk of suffering myocardial infarction. Thus, cGAMP can be used as a biomarker for myocardial infarction to assist in the diagnosis or risk prediction of myocardial infarction, providing a new approach for the diagnosis and risk prediction of myocardial infarction, and also providing a convenient and low-cost diagnostic tool for clinicians.

[0023] 2. cGAMP has a high diagnostic value as a single biomarker for diagnosing myocardial infarction. Specifically, when the cGAMP level is <1352 ng / L, the AUC is 0.66, the sensitivity is 56%, and the specificity is 68%; when the cGAMP level is ≥1352 ng / L, the AUC is 0.62, the sensitivity is 47%, and the specificity is 71%.

[0024] 3. The detection of the concentration of the biomarker cGAMP in the present invention can be completed through blood testing, can be implemented in routine clinical examinations, is easy to operate, and has high repeatability and reliability. It is suitable for large-scale promotion and application. Compared with traditional imaging examinations, it has the advantages of low cost and non-invasiveness, and improves patient acceptance and comfort. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 The U-shaped curve of cyclic dinucleotide cGAMP as a biomarker and the risk of myocardial infarction in patients with coronary heart disease;

[0026] Figure 2 The ROC curve constructed when cGAMP is used as a single biomarker to diagnose myocardial infarction when the cGAMP concentration of the coronary heart disease patient is ≥1352 ng / L in the embodiment;

[0027] Figure 3 This is the ROC curve constructed when cGAMP is used as a single biomarker to diagnose myocardial infarction when the cGAMP concentration of the coronary heart disease patient is less than 1352 ng / L in the example. DETAILED DESCRIPTION

[0028] The present invention is described in detail below with reference to the accompanying drawings and embodiments, but the present invention can be implemented in many different ways as limited and covered by the claims.

[0029] The experimental methods without specific conditions in the examples are usually carried out under conventional conditions or conditions recommended by the manufacturers. The materials and reagents used in the following examples are all commercially available unless otherwise specified.

[0030] Example

[0031] Validation of the risk prediction and diagnostic performance of a single biomarker, cGAMP, in myocardial infarction

[0032] 1.1 Case selection

[0033] Patients with coronary heart disease who were admitted to the Second Xiangya Hospital of Central South University for chest pain were selected. All patients were adults aged between 35 and 75 years old and had undergone coronary angiography to confirm whether they had myocardial infarction. In this embodiment, a total of 270 patients with coronary heart disease were selected.

[0034] 1.2 Case data

[0035] The relevant clinical background information of the patients was collected, including but not limited to the general condition of the patients (such as age, gender, weight, height, etc.), medical history (smoking history, drinking history, hypertension, diabetes, atrial fibrillation, etc.), laboratory tests (including blood routine, liver function, kidney function, electrolytes, blood lipids and other indicators), echocardiogram, medication use, etc. The specific statistical information of the case data is shown in Table 1.

[0036] 1.3 Detection of cyclic dinucleotide cGAMP value

[0037] Sample collection: Patients were given venous blood samples for subsequent laboratory testing. Blood samples were centrifuged at room temperature and plasma was collected for cGAMP concentration testing.

[0038] In the present invention, the plasma is separated by centrifugation at 2500 rpm for 5 minutes.

[0039] cGAMP detection: The cGAMP concentration in plasma was quantitatively analyzed using the human cyclic guanosine monophosphate adenosine monophosphate (cGAMP) enzyme-linked immunosorbent assay kit (provided by Fancovi, catalog number: F11293-A). The experimental operation was carried out according to the instructions of the kit, and the standard curve was established using a cGAMP standard solution of known concentration. The operation process ensured strict quality control and good experimental repeatability.

[0040] 1.4 Statistics

[0041] According to the cGAMP concentration in the plasma of patients with CHD, they were divided into three groups, namely Group 1, Group 2 and Group 3, and the baseline clinical data were compared. See Table 1 for details.

[0042] Table 1 Clinical data of cases

[0043]

[0044]

[0045]

[0046] 1.5 Data Analysis

[0047] In this example, among 270 patients, 212 (78.52%) were male, the median age was (61.83±10.78) years old, and the average plasma cGAMP concentration was (1352.58±106.02 ng / L).

[0048] As can be seen from Table 1, compared with patients in Group 2, more patients in Group 1 and Group 3 were diagnosed with myocardial infarction, and there were no significant differences in other variables. Specifically, 19 patients in Group 2 were diagnosed with myocardial infarction, 36 patients in Group 1 were diagnosed with myocardial infarction, and 34 patients in Group 3 were diagnosed with myocardial infarction. Among them, the myocardial infarction includes ST-segment elevation myocardial infarction (corresponding to ST-segment elevation myocardial infarction in Table 1) and non-ST-segment elevation myocardial infarction (corresponding to non-ST-segment elevation myocardial infarction in Table 1).

[0049] Of the 270 patients, 89 patients (32.96%) were diagnosed with myocardial infarction. The results of the univariate logistic regression model are shown in Table 2. Increased levels of HDL, apoA1, and LVEF were negatively correlated with the occurrence of MI, suggesting a protective effect. On the other hand, higher levels of free fatty acids, cTNT, NT-pro BNP, creatinine, and a history of hypertension were significantly positively correlated with an increased risk of myocardial infarction in people with coronary heart disease.

[0050] Table 2 Screening variables for univariate logistic regression model

[0051]

[0052] Please refer to Table 3. Taking Group 2 as the reference group, the risk of myocardial infarction in patients in Group 1 was 167% higher than that in Group 2 (adjusted OR: 2.67, 95% CI: 1.23-5.78, P=0.013); the risk of myocardial infarction in patients in Group 3 was 155% higher than that in Group 2 (adjusted OR: 2.55, 95% CI: 1.17-5.55, P=0.018).

[0053] Table 3 Relationship between cGAMP and myocardial infarction

[0054]

[0055] This trend is intuitively shown by the RCS curve, such as Figure 1 As shown, from Figure 1It can be seen that the concentration of the cyclic dinucleotide cGAMP in the plasma of patients with coronary heart disease shows a significant U-shaped relationship with the risk of suffering from myocardial infarction. When the concentration of cyclic dinucleotide cGAMP is around 1352 ng / L, it is predicted that the risk of myocardial infarction in patients with coronary heart disease is low; cGAMP concentrations that are too high or too low relative to the critical concentration (1352 ng / L) will increase the risk of myocardial infarction. When the concentration of cyclic dinucleotide cGAMP is extremely high (greater than 1450 ng / L) or extremely low (less than 1250 ng / L), it is predicted that the risk of myocardial infarction in patients with coronary heart disease is low or high.

[0056] That is, when the concentration of cyclic dinucleotide cGAMP = 1352 ng / L, the risk of myocardial infarction in patients with coronary heart disease is predicted to be low; when the concentration of cGAMP is <1352 ng / L, as the concentration of cyclic dinucleotide cGAMP in the plasma of patients with coronary heart disease increases, the risk of myocardial infarction in patients with coronary heart disease decreases (inverse correlation); when the concentration of cGAMP is ≥1352 ng / L, as the concentration of cyclic dinucleotide cGAMP in the plasma of patients with coronary heart disease increases, the risk of myocardial infarction in patients with coronary heart disease increases (positive correlation).

[0057] Cyclic dinucleotide cGAMP is used as a single biomarker to diagnose whether patients with coronary heart disease have myocardial infarction. The ROC curve analysis results are detailed in Figure 2 and Figure 3 ,from Figure 2 and Figure 3 It can be seen that regardless of whether the concentration of cGAMP in the plasma of patients with coronary heart disease is greater than or less than the critical concentration of 1352 ng / L, cGAMP has a moderate diagnostic ability for myocardial infarction. Specifically, when the cGAMP level is <1352 ng / L, the AUC is 0.66, the sensitivity is 56%, and the specificity is 68%; when the cGAMP level is ≥1352 ng / L, the AUC is 0.62, the sensitivity is 47%, and the specificity is 71%.

[0058] The above contents are further detailed descriptions of the present invention in combination with specific preferred embodiments, and it cannot be determined that the specific implementation of the present invention is limited to these descriptions. For ordinary technicians in the technical field to which the present invention belongs, several simple deductions and substitutions can be made without departing from the concept of the present invention, which should be regarded as belonging to the protection scope of the present invention.

Claims

1. Application of reagents for detecting the biomarker cyclic dinucleotide cGAMP in the preparation of diagnostic tools for myocardial infarction.

2. Application of biomarker cyclic dinucleotide cGAMP in the preparation of myocardial infarction risk prediction products.

3. The use according to claim 2, characterized in that: The risk of myocardial infarction can be predicted by detecting the concentration of cyclic dinucleotide cGAMP in the plasma of patients with coronary heart disease.

4. The use according to claim 3, characterized in that: The product for detecting the concentration of cyclic dinucleotide cGAMP in the plasma of patients with coronary heart disease is an ELISA kit.

5. The use according to claim 2, characterized in that: The myocardial infarction includes ST-segment elevation myocardial infarction and non-ST-segment elevation myocardial infarction.

6. The use according to any one of claims 2 to 5, characterized in that: The concentration of the cyclic dinucleotide cGAMP in the plasma of patients with coronary heart disease has a U-shaped relationship with the risk of myocardial infarction. A cGAMP concentration that is too high or too low relative to the critical concentration will increase the risk of myocardial infarction. The critical concentration is 1352 ng / L.

7. The use according to claim 6, characterized in that: When the concentration of cyclic dinucleotide cGAMP = 1352 ng / L, the risk of myocardial infarction in patients with coronary heart disease is predicted to be low; when the concentration of cyclic dinucleotide cGAMP is <1352 ng / L, the concentration of cyclic dinucleotide cGAMP is inversely correlated with the risk of myocardial infarction; when the concentration of cyclic dinucleotide cGAMP is >1352 ng / L, the concentration of cyclic dinucleotide cGAMP is positively correlated with the risk of myocardial infarction.

8. Application of the biomarker cyclic dinucleotide cGAMP in constructing a myocardial infarction risk prediction model.

9. The use according to claim 8, characterized in that: The input variables of the risk prediction model include the plasma concentration value of the cyclic dinucleotide cGAMP.

10. The use according to claim 9, characterized in that: The input variables of the risk prediction model also included age, sex, hypertension, diabetes, smoking, drinking, body mass index, systolic blood pressure, left ventricular ejection fraction, serum creatinine, cardiac troponin T, N-terminal pro-brain natriuretic peptide, high-density lipoprotein, apolipoprotein A1 and free fatty acids.

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