Application of SDF4 in screening reagents for early warning or treatment of aortic disease

Abstract

This invention discloses the application of SDF4 in screening agents for early warning or treatment of aortic disease, belonging to the field of disease biomarker technology. SDF4, as a targeted molecular biomarker for aortic disease, exhibits high specificity and sensitivity. Furthermore, inhibitors of SDF4 can be used to prepare agents for early warning and treatment of aortic disease, thus providing a new direction for clinical risk warning, early diagnosis, and drug treatment of aortic disease.

Description

Technical Field

[0001] This invention belongs to the field of disease biomarker technology, specifically relating to the application of SDF4 in screening reagents for early warning or treatment of aortic disease. Background Technology

[0002] Aortic disease is a critical cardiovascular emergency known for its high morbidity and mortality rates. It is broadly classified into thoracic aortic aneurysm, abdominal aortic aneurysm, and acute aortic syndrome. An aortic aneurysm is defined as an expansion of the aorta to a diameter more than 1.5 times the normal aortic diameter. Based on the location of involvement, it is often classified as thoracic aortic aneurysm or abdominal aortic aneurysm. Acute aortic syndrome mainly includes aortic dissection, intramural hematoma (IMH), and penetrating aortic ulcer. Most patients with this condition present with sudden, severe, tearing pain in the chest, back, or abdomen.

[0003] In recent years, aortic disease has shown a trend of affecting younger people, with a significantly higher incidence rate in men than women. The rapid increase in the incidence of aortic aneurysms / dissections is closely related to the lack of early diagnostic indicators. Currently, early diagnosis of aortic disease in clinical practice mainly relies on raising individual health awareness and combining examinations such as echocardiography and large vessel CTA to assess disease risk.

[0004] While aortic aneurysms / dissections can be monitored through regular imaging examinations to reflect the degree of aortic dilation, intervention criteria for aortic dilation are determined solely through evidence-based medicine surveys and lack scientific support. Furthermore, it is difficult to predict when a dilated aorta will rupture or its wall will tear, leading to dissection. Aortic aneurysms / dissections are often asymptomatic, with no warning signs. Studies have found that up to 60% of aortic dissection patients are not diagnosed before death. Even after clinical diagnosis, surgical intervention is the only option, which carries high risks, numerous complications, and a high mortality rate. The mortality rate within two weeks of onset can reach 80% for untreated patients, making treatment extremely challenging.

[0005] Aortic disease is closely related to genetic factors and variations in vascular tissue structure. It is difficult to assess using only parameters such as aortic diameter and hemodynamics, making this strategy unsuitable for screening, early warning, and prognosis in high-risk populations. Current research has identified biomarkers related to the diagnosis of different types of aortic aneurysms / dissections, such as D-dimer, high-sensitivity CRP, and hsTnT, which can aid in disease diagnosis. However, their specificity is poor, and they cannot be developed as targets for subsequent drug treatment. Patients with aortic dissection have elevated plasma D-dimer and high-sensitivity CRP, and approximately 50% of dissection patients have elevated peripheral blood high-sensitivity troponin T (hsTnT). However, these indicators are also elevated in patients with myocardial infarction, increasing the risk of misdiagnosis of aortic disease. Therefore, the indicators previously used in clinical differential diagnosis lack specificity and cannot meet the needs of early diagnosis of aortic disease.

[0006] Therefore, identifying early warning molecules and therapeutic targets for aortic disease and implementing early intervention is of great clinical significance for the treatment of aortic disease. Summary of the Invention

[0007] Technical problem solved: To address the above-mentioned technical problems, this invention provides the application of SDF4 in screening reagents for early warning and treatment of aortic diseases, which has high specificity and sensitivity, and provides a new direction for the use of SDF4 inhibitors in clinical aortic disease risk warning, early diagnosis and drug treatment.

[0008] Technical solution: Application of SDF4 in screening reagents for early warning or treatment of aortic diseases.

[0009] Application of SDF4 inhibitors in the preparation of reagents for the prevention or treatment of aortic diseases.

[0010] Preferably, the SDF4 inhibitor includes a calpain inhibitor and a pharmaceutically acceptable carrier.

[0011] Application of SDF4 in screening reagents for early warning or treatment of thoracic aortic aneurysms.

[0012] Application of SDF4 inhibitors in the preparation of reagents for the prevention or treatment of thoracic aortic aneurysms.

[0013] Preferably, the SDF4 inhibitor includes a calpain inhibitor and a pharmaceutically acceptable carrier.

[0014] Application of SDF4 in screening reagents for early warning or treatment of thoracic aortic dissection.

[0015] Application of SDF4 inhibitors in the preparation of reagents for the prevention or treatment of thoracic aortic dissection.

[0016] Preferably, the SDF4 inhibitor includes a calpain inhibitor and a pharmaceutically acceptable carrier.

[0017] Beneficial effects: Based on the lack of specific biomarkers for aortic aneurysm / dissection and the problem of intervention drug targets, this invention uses proteomics to screen for SDF4 protein that is specifically highly expressed in aortic dissection, and reduces SDF4 in animal models by applying calcium inhibitors, thereby achieving the therapeutic goal of improving the occurrence and development of aneurysm / dissection. Attached Figure Description

[0018] Figure 1 This is a volcano plot of differentially expressed proteins, where the horizontal axis represents the fold change (log2 value) of the differentially expressed proteins, and the vertical axis represents the p-value (-log2). 10 (Values), black dots represent proteins with no significant difference, red dots represent upregulated proteins, and green dots represent downregulated proteins; Figure 2 This is a bar chart of GO annotation results, where the horizontal axis represents the number of proteins and the vertical axis represents the annotated GO entries. Figure 3 The values ​​represent the peripheral blood SDF4 concentrations in patients with thoracic aortic dissection and healthy individuals. In this context, Control represents normal individuals, Aortic dissection represents patients with thoracic aortic dissection, and **** indicates P < 0.0001. Figure 4 This is the ROC curve for SDF4 prediction of thoracic aortic dissection. Figure 5 The Kaplan-Meier survival curves of mice in each group are shown, where Control: control group; Calpeptin: inhibitor group; BAPN: β-aminopropionitrile group; BAPN+Calpeptin: β-aminopropionitrile group combined with inhibitor group. Figure 6 The aortic dissection and rupture in mice in each group are as follows: Control: control group; Calpeptin: inhibitor group; BAPN: β-aminopropionitrile group; BAPN+Calpeptin: β-aminopropionitrile group combined with inhibitor group. Figure 7 This study compared the peripheral blood SDF4 concentration in two groups of mice: BAPN (fed only BAPN water) and BAPN+CAP (fed BAPN water and injected with Calpeptin). **P < 0.01. Implementation

[0019] The present invention will be further described below with reference to the accompanying drawings and specific embodiments.

[0020] Example 1: Screening of SDF4 as a biomarker Five patients with thoracic aortic dissection were randomly selected as the experimental group, and five healthy individuals were selected as the control group. Peripheral serum proteomics analysis was performed. TMT technology was used for protein quantification. By comparing protein samples in plasma samples from the thoracic aortic dissection group and the healthy control group, the ratio of the mean of all biologically replicated quantitative values ​​for each protein in the compared sample pair was taken as the fold change (FC). A t-test was used to determine the difference of each protein between the two samples, and the p-value was calculated. For this test, upregulated proteins were selected when FC ≥ 1.2 and p-value ≤ 0.05, and downregulated proteins were selected when FC ≤ 0.83 and p-value ≤ 0.05. A total of 1845 differentially expressed proteins were screened, of which 36 were upregulated and 16 were downregulated. Figure 1 ).

[0021] GO functional enrichment analysis of differentially expressed proteins was performed. Due to the large number of GO functional annotation entries, the top 10 entries in each major protein class were selected. Biological processes included proteolysis, ubiquitin-dependent proteolysis, lipid transport, immune responses, cell adhesion, homocellular adhesion via plasma membrane adhesion molecules, signal transduction, lipoprotein metabolism, proteolysis during cellular protein breakdown, and redox processes. Molecular functions included nucleic acid binding, serine endopeptidase activity, threonine endopeptidase activity, endopeptidase inhibitor activity, serine endopeptidase inhibitor activity, structural molecule activity, calcium ion binding, protein binding, ATP binding, and lipid binding. Emphasis was placed on proteins enriched in the calcium binding pathway and immune response pathways. Combined with the differential protein screening results, these proteins were co-enriched to SDF4 (P=0.0027). Figure 2 ).

[0022] Proteomics was used to identify SDF4, an immunomodulatory protein associated with calcium channels. Further analysis was conducted to verify the differential expression of SDF4 in peripheral serum between patients with thoracic aortic dissection and healthy controls. The concentration of SDF4 was detected using an ELISA kit. The results are shown in Table 1. The serum SDF4 concentration in patients with thoracic aortic dissection was 8.28 ± 0.41 ng / ml, while that in healthy controls was 2.12 ± 0.28 ng / ml (P < 0.0001), indicating a significant difference between the two groups (Table 1). Figure 3 ).

[0023] Table 1. Comparison of peripheral blood SDF4 concentrations between the two groups

[0024] Example 2: Application of SDF4 in screening early warning reagents for aortic disease To further explore the predictive value of peripheral blood SDF4 concentration for thoracic aortic dissection, ROC curves were plotted. The results showed that the area under the curve (AUC) for SDF4 in predicting thoracic aortic dissection was 0.959 (95% CI: 0.915–1.000, P < 0.0001), with a sensitivity of 97.5% and a specificity of 88.6%, indicating good predictive efficacy. (Results are as follows...) Figure 4 As shown.

[0025] Example 3: Application of SDF4 inhibitors in the preparation of therapeutic agents for aortic diseases To investigate the therapeutic effect of SDF4 as a drug target in aortic disease, a mouse aortic aneurysm / dissection model was constructed using a calpain inhibitor, and mouse survival, aneurysm formation / dissection rate were observed. In Experiment I, C57BL / 6 mice were selected and divided into four groups (Control group, Calpeptin group, BAPN group, and BAPN + Calpeptin group). The treatment measures were as follows: Control group: fed with double-distilled water for four weeks, and intraperitoneally injected with physiological saline three times a week at a dose of 0.2 ml / 10 g; Calpeptin group: fed with double-distilled water for four weeks, and intraperitoneally injected with diluted Calpeptin solution three times a week at a dose of 0.02 mg / 0.2 ml / 10 g; BAPN group: fed mice with BAPN at a dose of 1 g / kg / day, and intraperitoneally injected with physiological saline three times a week at a dose of 0.2 ml / 10 g; BAPN + Calpeptin group: fed mice with BAPN at a dose of 1 g / kg / day, and intraperitoneally injected with diluted Calpeptin solution three times a week at a dose of 0.02 mg / 0.2 ml / 10 g.

[0026] The results showed that the mean survival days for the Control group, Calpeptin group, BAPN group, and BAPN + Calpeptin group were 28.00 ± 0.00 days, 28.00 ± 0.00 days, 20.40 ± 1.43 days, and 24.50 ± 2.21 days, respectively. Further Kaplan-Meier survival curves were plotted for each group. Compared with the Control group and Calpeptin group, the survival of mice in the BAPN group was significantly reduced (Log Rank P < 0.0001). The survival of mice fed with BAPN and then injected intraperitoneally with Calpeptin was improved. The results are shown in Table 2 and [Table data missing]. Figure 5 .

[0027] Table 2. Aortic dissection formation and 28-day survival in mice of each group.

[0028] Note: Numerical data are expressed as mean ± standard deviation, and incidence is expressed as a percentage.

[0029] Gross dissection was performed to determine whether aortic dissection occurred in mice, and the occurrence and rupture were recorded. The incidence of thoracic aortic aneurysm / dissection (TAAD) in the Control group, Calpeptin group, BAPN group, and BAPN + Calpeptin group was 0%, 0%, 100% (5 cases of thoracic aortic aneurysm and 10 cases of thoracic aortic dissection out of 15 cases), and 67% (3 cases of thoracic aortic aneurysm and 7 cases of thoracic aortic dissection out of 15 cases), respectively. The rupture rates were 0%, 0%, 87%, and 13%, respectively. A bar chart was plotted to illustrate the formation and rupture of aortic dissection in mice. Figure 6 As shown in the figure. The results indicate that the application of calpain inhibitors can improve the occurrence of aortic aneurysm / dissection formation and dissection rupture.

[0030] Blood samples were collected from mice fed BAPN water and mice fed BAPN while simultaneously injected with the inhibitor Calpeptin. SDF4 concentration was detected using an ELISA kit. The results showed that the SDF4 concentration in the BAPN group was 19.47 ± 0.61 ng / ml, while the SDF4 concentration in the BAPN + Calpeptin group was 11.20 ± 5.03 ng / ml. The peripheral blood SDF4 concentration was significantly decreased in mice injected with the inhibitor (P < 0.05). These results suggest that calpain inhibitors can act as SDF4 inhibitors, reducing SDF4 concentration in peripheral blood and improving the formation of aortic disease. (Results are as follows...) Figure 7 As shown.

Claims

1. Application of SDF4 in the preparation of early warning reagent for thoracic aortic dissection.

2. Application of Calpeptin in the preparation of reagents for the treatment of thoracic aortic dissection.

Images