A biomarker for diagnosing and / or monitoring a large artery inflammation ischemic event, and applications and devices thereof

Abstract

The present application belongs to the technical field of diagnostic biomarkers, and particularly relates to a biomarker for diagnosing and / or monitoring ischemic events of Takayasu arteritis, application of the biomarker in preparation of a device for diagnosing and / or monitoring ischemic events of Takayasu arteritis, and a device for diagnosing and / or monitoring Takayasu arteritis comprising the biomarker. The present application discloses a biomarker for diagnosing and / or monitoring ischemic events of Takayasu arteritis, which comprises at least three of CD19+ B cell count, FABP3 level, leptin level, C4 level and IgA level. Compared with traditional vascular imaging evaluation, serum marker detection is more convenient, causes less damage to patients, and is free of radiation, and can not only dynamically monitor the degree of vascular damage and potential ischemic lesions of organs of a patient, but also precisely grasp possible biological processes in the body of the patient. The combination of multiple indexes can improve the accuracy of disease evaluation and avoid the risk of false positive or false negative in single index judgment.

Description

Technical Field

[0001] This invention belongs to the field of diagnostic biomarker technology, specifically relating to a biomarker for diagnosing and / or monitoring ischemic events in aortitis, its application in the preparation of a device for diagnosing and / or monitoring ischemic events in aortitis, a device for diagnosing and / or monitoring aortitis including the biomarker, and particularly relating to the application of the biomarker in the preparation of a device for diagnosing and / or monitoring ischemic adverse events related to severe stenosis in aortitis. Background Technology

[0002] Takayasu arteritis is a chronic, nonspecific, inflammatory disease of the large blood vessels, primarily affecting the aorta and its first-order branches, including the brachiocephalic trunk, common carotid artery, subclavian artery, and renal artery. Affected arteries exhibit thickened walls, narrowed lumens, and occlusion, leading to ischemic damage and even organ failure in vital organs, severely impacting the patient's life and health.

[0003] Adverse ischemic events in Takayasu arteritis refer to events related to ischemia in corresponding tissues and organs caused by severe stenosis or occlusion of affected blood vessels, such as cerebral ischemia, vision loss, renal atrophy, renal infarction, renal hypertension, pulmonary infarction, and myocardial infarction. In clinical practice, the examination of vascular lesions in Takayasu arteritis mainly relies on imaging examinations, including vascular ultrasound, CT, and MRI, lacking more convenient serological markers, making it difficult to accurately and dynamically assess vascular lesions. When the stenosis of affected blood vessels is greater than 50%, the related organs face the risk of ischemic damage. However, because the body's tissues and organs have a certain compensatory function, mild organ damage often does not have clinically noticeable manifestations or laboratory abnormalities, leading to delays in diagnosis and treatment. Therefore, it is particularly important to find biomarkers for ischemia in organs related to vascular stenosis in Takayasu arteritis.

[0004] The exploration of biomarkers for Takayasu arteritis has been a key focus for clinicians. Currently reported biomarkers are mostly associated with systemic inflammation, such as erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), amyloid A, and interleukin-6. After treatment with glucocorticoids and immunosuppressants, systemic inflammation is controlled, and these inflammatory markers gradually return to normal. However, long-term follow-up of Takayasu arteritis patients has revealed that in approximately 50% of patients, even after inflammation is controlled through treatment, vessel wall thickening continues to progress, leading to an increased risk of ischemia in corresponding tissues and organs, severely impacting long-term prognosis. Therefore, specific biomarkers for persistent vascular damage urgently need to be explored.

[0005] Takayasu arteritis primarily affects the adventitia, and its pathogenesis is extremely complex. During the pathogenesis of Takayasu arteritis, various immune cells and tissue cells participate in vascular lesions, including T lymphocytes, B lymphocytes, macrophages, and fibroblasts. Activated immune cells can secrete various pro-inflammatory and chemokines, such as IFN-γ, IL-17, CCL2, IL-16, YKL-40, and PTX3. Activated macrophages and tissue cells can secrete various matrix metalloproteinases (MMPs), such as MMP2, MMP3, and MMP9, which participate in vascular structural damage. Among these factors, some are related to systemic inflammation and are significantly suppressed after immunosuppressive therapy; however, it remains unclear which factors participate in persistent vascular damage and can predict organ ischemia risk. Summary of the Invention

[0006] Therefore, the present invention aims to provide a biomarker for diagnosing and / or monitoring ischemic events in aortitis, the application of its preparation in a device for diagnosing and / or monitoring ischemic events in aortitis, a device for diagnosing and / or monitoring aortitis including the biomarker, and a combination of multiple factors to predict ischemic adverse events related to severe vascular stenosis in aortitis.

[0007] To achieve the above objectives, the solution adopted by the present invention is as follows:

[0008] In a first aspect, the present invention provides a biomarker for diagnosing and / or monitoring Takayasu arteritis, comprising at least three of the following: CD19+ B cell count, FABP3 level, leptin level, and C4 and IgA levels.

[0009] Preferably, the biomarker uses human serum, plasma, or blood as a biological sample.

[0010] In a second aspect, the present invention also provides a device for the diagnosis and / or monitoring of aortitis, including the biomarkers described above.

[0011] Preferably, the device includes a diagnostic chip and / or a diagnostic kit.

[0012] Thirdly, the present invention also provides the use of the biomarkers described above in the preparation of a device for diagnosing and / or monitoring ischemic events of aortitis, detecting at least three of the following in human serum, plasma or blood: CD19+ B cell count, FABP3 level, leptin level, and C4 and IgA levels.

[0013] Preferably, it is used in the preparation of a device for diagnosing and / or monitoring ischemic adverse events of aortitis.

[0014] Preferably, the detection is performed on human serum, plasma, or blood in the following ways:

[0015] The number of CD19+B cells is greater than or equal to 300 cells / µl.

[0016] and / or FABP3 levels greater than 80 pg / ml

[0017] and / or leptin levels greater than 29 ng / ml,

[0018] and / or C4 levels not greater than 0.24 g / L,

[0019] And / or IgA levels not greater than 2.20 g / L.

[0020] Preferably, at least three of the following (a), (b), (c), and (d) are detected in human serum, plasma, or blood:

[0021] (a) CD19+ B cell count greater than or equal to 300 cells / µl

[0022] (b) FABP3 level greater than 80 pg / ml

[0023] (c) Leptin levels greater than 29 ng / ml

[0024] (d) C4 level not greater than 0.24 g / L,

[0025] (e) IgA levels not exceeding 2.20 g / L,

[0026] This suggests a significantly increased risk of ischemic adverse events associated with severe stenosis of large vessels due to Takayasu arteritis.

[0027] The pathogenesis of Takayasu arteritis is complex, involving intertwined processes at different stages, including systemic inflammation, vascular inflammation, vascular damage and remodeling, and organ ischemia. During this process, various cytokines undergo dynamic changes. In the systemic inflammation stage, multiple inflammatory factors, such as immunoglobulins and complement, rise, but these factors can be quickly controlled and show a downward trend after treatment. However, factors causing persistent vascular damage are often uncontrollable by existing treatment regimens and persist even after systemic inflammation subsides, ultimately leading to organ ischemia. Therefore, combining multiple factors may more accurately predict the risk of severe vascular stenosis and ischemic adverse events in Takayasu arteritis.

[0028] According to the present invention, the inventors have discovered that B lymphocytes may be important cells involved in the pathogenesis of Takayasu arteritis. After activation, B cells differentiate into plasma cells, which are the main source of immunoglobulins such as IgG, IgA, and IgM. Furthermore, research has shown that perivascular adipose tissue also plays an important role in the vascular lesions of Takayasu arteritis, and its secreted adipokines, such as leptin, may promote adventitia damage by regulating the function of adventitia fibroblasts. Therefore, this invention discloses the value and clinical application of combining CD19+ B cell count, FABP3, leptin, C4, and IgA in predicting ischemic adverse events in Takayasu arteritis.

[0029] B lymphocytes are immune cells involved in adaptive immune responses, with CD19 as a marker. In clinical practice, flow cytometry can be used to detect CD19+ cells to monitor the number and proportion of B cells. Current research has found that the proportion of CD19+ B cells in patients with Takayasu arteritis is significantly higher than in healthy controls, and CD19+ B cell infiltration is also present in the local vascular lesions. Studies have reported that the B cell activation molecules BAFF and APRIL are significantly higher in patients with Takayasu arteritis than in healthy controls. In addition, current research has found that IgG levels in patients with Takayasu arteritis are positively correlated with disease activity, and plasma cell infiltration and IgG expression are also present in the local lesions of the affected vessels. Therefore, CD19+ B cells may be closely related to the vascular lesions of Takayasu arteritis.

[0030] FABP3 is an intracellular lipid-binding protein whose functions are involved in various biological processes, including fatty acid uptake, transport, and metabolic regulation. In Takayasu arteritis, vascular lesions are predominantly located in the adventitia, and adventitia fibroblasts are the main cells of the arterial adventitia; their dysfunction is a significant cause of vascular lesions in Takayasu arteritis. Our previous studies found that FABP3 expression was significantly increased in adventitia fibroblasts of Takayasu arteritis patients. In vitro, FABP3 can promote extracellular matrix secretion by regulating fatty acid oxidation in adventitia fibroblasts, thus participating in the development of vascular fibrosis. Therefore, FABP3 may be involved in the vascular fibrosis process in Takayasu arteritis.

[0031] Leptin is an adipokine primarily expressed by mature adipocytes. It mainly regulates the body's perception of fat and controls food intake by acting on the central nervous system. However, in pathological conditions, leptin can also regulate the function of other cells by acting on various peripheral tissues. Early studies found that high leptin levels (>28.9 ng / ml) in patients with Takayasu arteritis were associated with the progression of affected vessel wall structures. Therefore, leptin may be related to the severity of vascular lesions in Takayasu arteritis and may have some value in predicting ischemic adverse events related to vascular stenosis.

[0032] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0033] Compared to traditional vascular imaging assessments, serum biomarker testing is more convenient, less invasive to patients, and radiation-free. It can not only dynamically monitor the degree of vascular damage and potential organ ischemia, but also provide a precise understanding of possible biological processes within the patient's body. Combining multiple indicators can improve the accuracy of disease assessment and avoid the risk of false positives or false negatives from relying on a single indicator. Attached Figure Description

[0034] Figure 1 This is a graph showing the correlation analysis between different indicators in patients with aortitis in an embodiment of the present invention. Detailed Implementation

[0035] This invention provides a multifactorial biomarker for diagnosing and / or monitoring ischemic events in large vessel arteritis and predicting the risk of ischemic events in patients.

[0036] The technical solution of the present invention will be further described below with reference to specific embodiments; however, the scope of protection of the present invention is not limited to these embodiments. All changes or equivalent substitutions that do not depart from the concept of the present invention are included within the scope of protection of the present invention.

[0037] Example:

[0038] This invention analyzed the levels of various inflammatory markers, cytokines, and different immune cells in the peripheral blood of 32 patients with Takayasu arteritis (Table 1), and investigated the relationship between these markers and severe stenosis of affected vessels and organ ischemia events in Takayasu arteritis, aiming to identify potential biomarkers for predicting ischemic adverse events in Takayasu arteritis. Patient general information, clinical characteristics, and laboratory test results are shown in Table 1.

[0039] Table 1 General information and clinical characteristics of patients with Takayasu arteritis

[0040]

[0041]

[0042] #Ischemic type: 7 cases of renovascular renal failure, 1 case of visual impairment, 6 cases of cerebral infarction, and 1 case of pulmonary hypertension with heart failure.

[0043] The differences in different laboratory indicators among patients with organ ischemia, ischemic type, and severe vascular stenosis were analyzed (Table 2). The results showed that the number of CD19+ B cells, serum FABP3, and leptin levels were significantly higher in patients with organ ischemic events than in patients without organ ischemic events. The number of CD19+ B cells, serum FABP3, and leptin levels in ischemic patients were also significantly higher than in patients with vascular inflammation. The number of B cells in patients with severe stenosis was significantly higher than in patients without severe stenosis, and the levels of FABP3 and leptin also showed an upward trend.

[0044] Table 2. Differences in laboratory indicators among patients with different clinical features of Takayasu arteritis

[0045]

[0046]

[0047] Through analysis such as Figure 1 The correlations of different indicators showed that CD19+ B cell count and leptin were positively correlated with various immune activation indicators or chemokines, such as MMP3, CCL2, YKL40, and IL-16, all of which are associated with vascular damage. In addition, leptin was negatively correlated with ESR, IgG, and IgA, suggesting that it may be associated with vascular lesions after the body's inflammation subsides.

[0048] Peripheral blood CD19+ B cell count, FABP3, leptin, IgA, and C4 levels were used to predict organ damage related to vascular stenosis in patients with aortitis. The sensitivity, specificity, area under the ROC curve, and corresponding cutoff values ​​are shown in Table 5.

[0049] Table 5. The efficacy of different indicators in predicting organ ischemia in patients with Takayasu arteritis.

[0050]

[0051] In the table, severe stenosis is defined as stenosis >75%.

[0052] Predictive Model 1: For patients without whole-body imaging examinations:

[0053] By jointly predicting the above five peripheral blood indicators, ①B cell count ≥300 cells / µL, ②leptin level >29 ng / ml, ③FABP3 >80 pg / ml, ④IgA ≤2.2 g / L, and ⑤C4 ≤0.24 g / L were each assigned a score of 1. The analysis showed that when the total score was ≥3 points, that is, when at least three of the above conditions were met, the sensitivity for predicting ischemic adverse events was 78.57%, the specificity was 88.89%, and the area under the curve was 0.88 (p<0.001).

[0054] Predictive Model 2: For patients with whole-body imaging results:

[0055] Since severe vascular stenosis is a direct cause of organ and tissue ischemia, this invention also analyzed the predictive value of the number of severely stenotic vessels (>75%) in patients for ischemic events. It was found that the sensitivity of predicting ischemic events with two or more severely stenotic vessels was 71.43%, the specificity was 66.67%, and the area under the curve was 0.7 (p = 0.03, Table 5). The imaging indicators were combined with the above five peripheral blood indicators for prediction: ① number of vessels with severe stenosis (>75%) ≥2, ② B cell count ≥300 / μL, ③ leptin level >29 ng / ml, ④ FABP3 >80 pg / ml, ⑤ IgA ≤2.2 g / L, ⑥ C4 ≤0.24 g / L. Each was assigned a score of 1. Further analysis revealed that when the total score was ≥3, i.e., at least three of the above conditions were met, the sensitivity for predicting ischemic adverse events was 85.7%, the specificity was 83.33%, and the area under the curve was 0.90 (p<0.001). These results show that using the number of vessels with severe stenosis from imaging alone is not a good predictor of ischemic events, while combining serological indicators can significantly improve predictive efficacy.

[0056] Application Examples:

[0057] In clinical practice, CD19+ B cell counts are detected by flow cytometry, and serum FABP3, leptin, IgA, and C4 levels are detected by enzyme-linked immunosorbent assay (ELISA). By analyzing the levels of these indicators in patients, scores are assigned according to Model 1 criteria. A total score ≥3 points predicts a higher risk of vascular injury and organ ischemia. In the following cases, patients 1-3 scored 5, 4, and 4 points respectively, and all experienced ischemic adverse events. Case 4 had a predicted score of 1 point, without severe stenosis or occlusion of blood vessels, and no ischemic adverse events occurred (Table 6).

[0058] Table 6. Specific Implementation Cases of the Predictive Model for Ischemic Events in Patients with Takayasu's Arteritis

[0059]

[0060]

[0061] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A biomarker for diagnosing and / or monitoring ischemic events in large vessel arteritis, characterized in that, It consists of CD19+ B cell count, FABP3 level, leptin level, C4 level, and IgA level.

2. The biomarker for diagnosing and / or monitoring Takayasu arteritis according to claim 1, characterized in that, The biomarkers utilize human blood as a biological sample.

3. A device for diagnosing and / or monitoring aortitis, comprising the biomarkers described in claim 1 or 2.

4. The apparatus according to claim 3, characterized in that, The device includes a diagnostic chip and / or a diagnostic kit.

5. The use of a biomarker in the preparation of a device for diagnosing and / or monitoring ischemic events in aortitis, characterized in that, The device is used to detect biomarkers in human blood, including at least three of the following: CD19+ B cell count, FABP3 level, leptin level, C4 level, and IgA level.

6. The application according to claim 5, characterized in that, Its application is in the preparation of devices for the diagnosis and / or monitoring of ischemic adverse events associated with severe stenosis of large vessels in Takayasu arteritis.

7. The application according to any one of claims 5-6, characterized in that, Detection of: in human blood The number of CD19+B cells is greater than or equal to 300 cells / µl. and / or FABP3 levels greater than 80 pg / ml and / or leptin levels greater than 29 ng / ml, and / or C4 levels not greater than 0.24 g / L, And / or IgA levels not greater than 2.20 g / L.

8. The application according to claim 7, characterized in that, Detect at least three of the following in human blood: (a), (b), (c), (d), and (e): (a) CD19+ B cell count greater than or equal to 300 cells / µl (b) FABP3 level greater than 80 pg / ml (c) Leptin levels greater than 29 ng / ml, (d) C4 level not greater than 0.24 g / L, (e) IgA levels not exceeding 2.20 g / L, This suggests a significantly increased risk of ischemic adverse events associated with severe stenosis of large vessels due to Takayasu arteritis.

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