Application of biomarker cyclic dinucleotide cGAMP in diagnosis of coronary artery three-branch lesion

Abstract

The invention provides an application of a biomarker cyclic dinucleotide cGAMP in diagnosis of coronary artery three-branch lesions. The application specifically relates to an application of the biomarker cyclic dinucleotide cGAMP in preparation of a product for diagnosing the three coronary artery diseases and an application of the biomarker cyclic dinucleotide cGAMP in construction of a model for diagnosing the three coronary artery diseases. Experimental results show that cGAMP concentrations of three coronary artery disease patients and non-three coronary artery disease patients in coronary heart disease patients are differentially expressed, the cGAMP as a single biomarker for diagnosing the three coronary artery diseases has high diagnostic value, and specifically, AUC is 0.71, sensitivity is 73%, specificity is 62%, and accuracy is 68%.

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 of three-vessel coronary artery lesions. Background Art

[0002] Triple-vessel disease (TSD) is a serious type of coronary heart disease, usually manifested by varying degrees of stenosis or occlusion of the three main coronary arteries (right coronary artery, left anterior descending coronary artery, and left circumflex coronary artery). Clinical symptoms (such as chest pain and dyspnea) and physical examinations are common methods for early screening of coronary heart disease, but they have great limitations. Many patients with triple-vessel coronary artery disease may not have obvious clinical symptoms in the early stages of the disease, especially those in the asymptomatic high-risk group, which makes it difficult to evaluate clinical symptoms comprehensively and accurately. In addition, the clinical manifestations of coronary heart disease vary greatly from person to person, and many symptoms may be similar to those of other diseases (such as gastrointestinal diseases or respiratory diseases), which affects the early detection of the disease.

[0003] At present, the prediction of three-vessel coronary artery disease mainly relies on clinical symptom assessment, imaging examination and joint analysis of certain blood markers. Among them, the manifestation of clinical symptoms has large individual differences, and in the early stage of three-vessel coronary artery disease, patients may not show obvious symptoms. Therefore, relying on symptom assessment may lead to missed diagnosis or misdiagnosis, especially in asymptomatic high-risk patients. Imaging examination is currently a routine means of diagnosing three-vessel coronary artery disease, among which coronary angiography is widely regarded as the gold standard for diagnosis. Although imaging examination has high diagnostic accuracy, it is expensive and requires high-end equipment support. Some examinations (such as coronary angiography) are invasive operations, which may pose certain risks to patients. In addition, the diagnostic results of imaging examinations may be affected by factors such as equipment accuracy and examination timing, and cannot provide early warning of the occurrence of the disease, so it cannot meet the needs of early, convenient and efficient diagnosis. Traditional blood markers (such as high-sensitivity C-reactive protein, blood lipids, etc.) can help reflect systemic inflammation or metabolic abnormalities of coronary heart disease in some cases, but their specificity is poor, it is difficult to distinguish different types of coronary artery lesions, and they cannot provide sufficient accuracy to diagnose the occurrence of three-vessel coronary artery disease.

[0004] Triple coronary artery disease not only increases the risk of acute coronary syndrome (ACS) in patients, but is also associated with a poor prognosis. Early and accurate prediction is of great significance for formulating appropriate treatment plans and improving the long-term prognosis of patients. Summary of the invention

[0005] The purpose of the present invention is to provide a new biomarker cyclic dinucleotide cGAMP for auxiliary diagnosis of three-vessel coronary artery lesions, thereby providing a new approach for the diagnosis of three-vessel coronary artery lesions.

[0006] In order to achieve the above-mentioned object, the present invention provides the use of cyclic dinucleotide cGAMP as a biomarker in the preparation of a product for diagnosing triple coronary artery lesions.

[0007] In a specific embodiment, the cyclic dinucleotide cGAMP is a plasma marker.

[0008] In a specific embodiment, the diagnostic product is a reagent or a kit.

[0009] In a specific embodiment, the test sample of the diagnostic product is plasma.

[0010] In a specific embodiment, when the cGAMP concentration in the plasma of a patient with coronary heart disease is greater than a critical value of the cGAMP concentration, the diagnostic rate for diagnosing the patient with coronary heart disease as having three-vessel coronary artery disease is high, wherein the critical value of the cGAMP concentration is 1375.479 pg / mL.

[0011] In a specific embodiment, when the cGAMP concentration in the plasma of a patient with coronary heart disease is less than the critical value of the cGAMP concentration, the diagnostic rate of diagnosing the patient with coronary heart disease as having three-vessel coronary artery disease is low, wherein the critical value of the cGAMP concentration is 1375.479 pg / mL.

[0012] In a specific embodiment, the cGAMP concentration value in the blood of patients with coronary heart disease is positively correlated with the diagnosis rate of three-vessel coronary artery lesions.

[0013] In the present invention, the diagnosis of triple coronary artery disease by the biomarker cyclic dinucleotide cGAMP includes the following steps: step 1), using an anticoagulant tube to collect venous blood from fasting patients; step 2), separating plasma from the venous blood and performing cGAMP quantitative detection; step 3), based on the cGAMP quantitative detection result or the combination of the cGAMP quantitative detection result and other clinical variables, diagnosing whether the coronary heart disease patient has triple coronary artery disease, wherein the other clinical variables include age, the presence or absence of diabetes, left ventricular short-axis shortening rate, creatine kinase isoenzyme, and fasting blood glucose.

[0014] Preferably, an ELISA kit is used for cGAMP quantitative detection.

[0015] Preferably, when the cGAMP concentration value in the plasma of a patient with coronary heart disease is greater than the critical value of the cGAMP concentration, the diagnostic rate for diagnosing the patient with coronary heart disease as having three-vessel coronary artery lesions is high; when the cGAMP concentration value in the plasma of a patient with coronary heart disease is less than the critical value of the cGAMP concentration, the diagnostic rate for diagnosing the patient with coronary heart disease as having three-vessel coronary artery lesions is low, wherein the critical value of the cGAMP concentration is 1375.479 pg / mL.

[0016] The present invention also provides the application of the biomarker cyclic dinucleotide cGAMP in constructing a diagnostic model for three-vessel coronary artery lesions.

[0017] In a specific embodiment, the input variables of the diagnostic model include the concentration value of the cyclic dinucleotide cGAMP.

[0018] In a specific embodiment, the input variables of the diagnostic model further include age, the presence or absence of diabetes, left ventricular fractional shortening, creatine kinase isoenzyme and fasting blood glucose.

[0019] The beneficial effects of the present invention include at least:

[0020] 1. The present invention discovers for the first time that there is a difference in the expression of cGAMP concentration between patients with coronary heart disease and those without three-vessel coronary artery lesions. Thus, cGAMP can be used as a biomarker for three-vessel coronary artery lesions to assist in the diagnosis or prognosis prediction of three-vessel coronary artery lesions, providing a new approach for the diagnosis and prognosis prediction of three-vessel coronary artery lesions, and also providing clinicians with a convenient and low-cost diagnostic tool.

[0021] 2. cGAMP has a high diagnostic value as a single biomarker for diagnosing triple coronary artery disease. Specifically, the AUC is 0.71, the sensitivity is 73%, the specificity is 62%, and the accuracy is 68%. When cGAMP is combined with multiple clinical variables (age, presence or absence of diabetes, left ventricular short-axis shortening rate, creatine kinase isoenzyme, and fasting blood glucose), the prediction accuracy can be significantly improved. The established ROC curve and nomogram can provide clinicians with convenient and accurate tools, which helps to identify high-risk patients early and formulate personalized treatment plans.

[0022] 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 advantage of low cost. Compared with traditional coronary artery examinations, it has the advantage of being non-invasive, which improves patient acceptance and comfort. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1Violin plots of cGAMP distribution of patients with three-vessel coronary artery lesions and patients with non-branch lesions in Example 1;

[0024] Figure 2 The ROC curve constructed for the diagnosis of three-vessel coronary artery disease using cGAMP as a biomarker in Example 1;

[0025] Figure 3 The ROC curve constructed for diagnosing three-vessel coronary artery disease by combining the biomarker cGAMP in Example 2 with multiple clinical variables (including age, presence or absence of diabetes, left ventricular fractional shortening, creatine kinase isoenzymes, and fasting blood glucose);

[0026] Figure 4 Based on Example 2 Figure 3 The nomogram for constructing the ROC curve. DETAILED DESCRIPTION

[0027] 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.

[0028] 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.

[0029] Example 1

[0030] Validation of the diagnostic performance of a single biomarker, cGAMP, in the diagnosis of triple-vessel coronary artery disease

[0031] 1.1 Case selection

[0032] Patients with chronic coronary syndrome were selected, including patients with long-term angina pectoris, chest pain and other symptoms. All patients were adults aged between 35 and 75 years old, and had undergone coronary angiography to confirm whether there were three-vessel coronary artery lesions. In this embodiment, a total of 506 patients with coronary heart disease were selected and divided into two groups according to whether there were three-vessel coronary artery lesions, specifically: 241 cases in the three-vessel lesion group and 264 cases in the non-three-vessel lesion group.

[0033] 1.2 Case data

[0034] The relevant clinical background information of the patients was collected, including but not limited to the following: (1) Age: the actual age of the patients, calculated in years; (2) History of diabetes: based on the medical records of the patients, determine whether there is a history of diabetes (diabetes vs. no diabetes); (3) Fractional shortening of the left ventricle (FS%): measured by echocardiography, using conventional cardiac ultrasound examination to measure the fractional shortening of the left ventricle; (4) Creatine kinase isoenzyme: the CK-MB level in the patient's serum was measured by blood test; (5) Fasting blood glucose: the blood glucose level of the patient was measured after fasting for 12 hours (unit: mg / dL).

[0035] According to the presence or absence of three-vessel coronary artery lesions, the baseline clinical data of the three-vessel lesion group and the non-three-vessel lesion group were compared. See Table 1 for details.

[0036] Table 1 Clinical data of cases

[0037]

[0038]

[0039] 1.3 Detection of cyclic dinucleotide cGAMP value

[0040] 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.

[0041] In the present invention, the plasma is separated by centrifugation at 2500 rpm for 5 minutes.

[0042] 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.

[0043] 1.4 Data Statistical Analysis

[0044] In the present invention, the average cGAMP concentration of the three-vessel lesion group was 1370.84±105.57pg / mL, and the average concentration of the non-three-vessel lesion group was 1325.36±109.40pg / mL. The statistical analysis results showed that the t value was -4.75 and the P value was less than 0.001, indicating that the difference in cGAMP concentration between the two groups was statistically significant.

[0045] Violin plots are used to show the differences in cGAMP distribution in patients with and without three-vessel coronary artery disease. For details, see Figure 1, This figure shows the distribution difference of cGAMP concentration between the three-vessel coronary artery lesions (lesions vs. no lesions) groups, and specifically shows the data distribution pattern, median, quartiles and outliers of each group. Among them, the X-axis: indicates whether the patient has three-vessel coronary artery lesions, divided into two groups: "three-vessel lesions" and "no three-vessel lesions"; the Y-axis: indicates the cGAMP concentration, the unit is ng / mL. Figure 1 It can be seen that the cGAMP level in patients with triple coronary artery lesions is significantly higher than that in patients without triple coronary artery lesions, which verifies the potential of cGAMP in the diagnosis of triple coronary artery lesions.

[0046] The ROC curve analysis results are shown in Figure 2 ,from Figure 2 It can be seen that:

[0047] (1) AUC (area under the curve): the AUC value was 0.71 (95% confidence interval: 0.67-0.76). This result indicated that cGAMP as a single biomarker had moderate diagnostic performance in distinguishing the presence or absence of triple coronary artery lesions.

[0048] (2) Sensitivity: 73% (95% CI: 67%-79%), indicating that cGAMP can correctly identify most positive cases when detecting three-vessel lesions.

[0049] (3) Specificity: 62% (95% CI: 57%-68%), indicating that cGAMP has a certain accuracy in identifying patients with non-three-vessel lesions.

[0050] (4) Accuracy: 68% (95% CI: 63%-72%), that is, the method was able to correctly classify 68% of the patients.

[0051] (5) Positive predictive value (PPV) and negative predictive value (NPV): Positive predictive value (PPV): 64% (95% CI: 58%-70%), that is, when the cGAMP concentration is higher than a certain critical value, the probability that the patient does have three-vessel lesions is 64%; Negative predictive value (NPV): 72% (95% CI: 66%-78%), that is, when the cGAMP concentration is lower than the critical value, the probability that the patient does not have three-vessel lesions is 72%.

[0052] (6) Critical value: When the cGAMP concentration was 1375.479 pg / mL, the ROC curve performed best, with the best balance between sensitivity and specificity.

[0053] Example 2

[0054] A diagnostic model combining the biomarker cGAMP with multiple clinical variables

[0055] (1) Multivariate ROC curve: A multivariate prediction model was constructed by combining cGAMP concentration values ​​and other clinical variables, including age, the presence or absence of diabetes, left ventricular fractional shortening, creatine kinase isoenzymes, and fasting blood glucose. Figure 3 The ROC curve of the model was shown, with an AUC value of 0.80 (95% confidence interval: 0.72-0.88), indicating that the model has high diagnostic accuracy and can effectively distinguish patients with triple coronary artery lesions.

[0056] (2) Construction and application of the nomenclature: Figure 3 The ROC curve results further constructed a nomogram, such as Figure 4 As shown in the figure, this nomogram helps clinicians quickly assess the risk of three-vessel coronary artery disease in patients by assigning specific scores to each clinical variable (including cGAMP). The specific approach is to accumulate the scores of each variable to obtain the patient's risk score, which in turn helps doctors make decisions on whether to further treat or intervene.

[0057] It should be noted that the experimental data of the diagnostic model of Example 2 is the same as the experimental data of Example 1, the only difference being that Example 1 is a univariate diagnostic model and Example 2 is a multivariate diagnostic model.

[0058] Combining Example 1 and Example 2, it can be seen that cGAMP has a high diagnostic value as a single biomarker for diagnosing triple coronary artery disease. Specifically, the AUC is 0.71, the sensitivity is 73%, the specificity is 62%, and the accuracy is 68%. After cGAMP is combined with multiple clinical variables, the prediction accuracy can be significantly improved. The established ROC curve and nomogram can provide clinicians with convenient and accurate tools, which are helpful for early identification of high-risk patients and the formulation of personalized treatment plans.

[0059] 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 biomarker cyclic dinucleotide cGAMP in the preparation of diagnostic products for triple coronary artery lesions.

2. The use according to claim 1, characterized in that: The cyclic dinucleotide cGAMP is a plasma marker.

3. The use according to claim 1, characterized in that: The diagnostic product is a reagent or a kit.

4. The use according to claim 1, characterized in that: The test sample of the diagnostic product is plasma.

5. The use according to claim 4, characterized in that: When the cGAMP concentration value in the plasma of a patient with coronary heart disease is greater than the critical value of the cGAMP concentration, the diagnostic rate of diagnosing the patient with coronary heart disease as having three-vessel coronary artery lesions is high, wherein the critical value of the cGAMP concentration is 1375.479 pg / mL.

6. The use according to claim 4 or 5, characterized in that: When the cGAMP concentration value in the plasma of a patient with coronary heart disease is less than the critical value of the cGAMP concentration, the diagnostic rate of diagnosing the patient with coronary heart disease as having three-vessel coronary artery lesions is low, wherein the critical value of the cGAMP concentration is 1375.479 pg / mL.

7. The use according to claim 5, characterized in that: The cGAMP concentration in the blood of patients with coronary heart disease is positively correlated with the diagnosis rate of three-vessel coronary artery lesions.

8. Application of the biomarker cyclic dinucleotide cGAMP in constructing a diagnostic model for triple-vessel coronary artery disease.

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

10. The use according to claim 9, characterized in that: The input variables of the diagnostic model also include age, the presence or absence of diabetes, left ventricular fractional shortening, creatine kinase isoenzyme and fasting blood glucose.

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