Plasma biomarkers for early assessment of the degree of myocardial fibroblast activation in patients with acute myocardial infarction and use thereof

Abstract

This application discloses a plasma biomarker for early assessment of myocardial fibroblast activation in patients with acute myocardial infarction (AMI) and its application. This application is the first to propose HBP as a biomarker for assessing the activation level of myocardial fibroblasts in AMI patients, aiming to utilize HBP levels for effective early assessment of myocardial fibroblast activation in AMI patients, ensuring timely clinical intervention and ultimately improving the prognosis of these high-risk patients. The biomarker (HBP) provided in this application enables non-invasive in vivo monitoring of the dynamic activity of myocardial fibrosis, overcoming the limitations of traditional imaging techniques such as late gadolinium enhancement in capturing the real-time activation status of fibroblasts during the critical repair period after myocardial infarction, and has promising clinical application prospects.

Description

Technical Field

[0001] This application relates to a plasma biomarker for early assessment of the activation level of myocardial fibroblasts in patients with acute myocardial infarction and its application, belonging to the field of biomedical detection technology. Background Technology

[0002] Heart failure (HF) is the end stage of various heart diseases and one of the leading causes of death and hospitalization worldwide. With the development of chest pain centers and the widespread use of reperfusion therapy in my country, the acute mortality rate of patients with acute myocardial infarction (AMI) has significantly decreased. However, the proportion of survivors who develop adverse ventricular remodeling and eventually progress to HF remains high. In the pathophysiological mechanism of HF, myocardial fibrosis driven by fibroblast activation is a key step. Excessive deposition of collagen fibers in the damaged heart leads to myocardial fibrosis. This process not only increases ventricular wall stiffness and decreases compliance but also disrupts the continuity of myocardial electrical conduction, increasing the risk of malignant arrhythmias.

[0003] However, myocardial fibrosis is not a single pathological process, but rather highly heterogeneous and dynamic. Based on its mechanism and morphological characteristics, it is mainly divided into two categories: (1) reparative fibrosis: mainly occurs in the infarcted area. Due to myocardial cell necrosis, fibroblasts are activated and secrete collagen to form scar tissue to replace necrotic cells and prevent ventricular rupture. This is the body's natural protective response; (2) reactive fibrosis: mainly occurs in the non-infarcted area. It manifests as diffuse collagen fiber deposition, mainly distributed in the myocardial interstitium and around blood vessels. This fibrosis is not for the purpose of replacing necrotic cells, but rather a pathological response of the heart to chronic inflammation, neurohumoral activation, or pressure load. Although reparative fibrosis is necessary in the early post-myocardial infarction period, excessive, persistent, and diffuse reactive fibrosis is a key factor leading to ventricular dysfunction, structural remodeling, and the development of heart failure. Therefore, there is an urgent clinical need for a detection method that can identify the degree of fibrosis activation.

[0004] Currently, the main clinical methods for assessing myocardial fibrosis include invasive endocardial biopsy (EMB) and non-invasive cardiac magnetic resonance imaging (CMR), but both have technical limitations in assessing fibrotic activity. While EMB is the gold standard for quantifying collagen deposition, myocardial fibrosis, especially reactive fibrosis, is typically patchy or heterogeneous. The tissue samples obtained by EMB are extremely small (approximately 1-2 mm³), making it difficult to represent the pathological state of the entire left ventricle, and single-point biopsies often lead to serious misjudgments of the degree of fibrosis. Furthermore, EMB is an invasive procedure with risks of serious complications such as cardiac perforation, cardiac tamponade, and conduction block. For patients in the acute phase of acute myocardial infarction (AMI), EMB is considered a contraindication due to its extremely high operational risks.

[0005] Late gadolinium enhancement (LGE) technology in cardiac MRI relies on the difference in contrast agent elution rates between necrotic / scar tissue and normal myocardium. It is well-suited for detecting focal, dense, repair-related fibrosis and is considered the gold standard for assessing myocardial scarring and irreversible fibrosis. However, for diffuse reactive fibrosis distributed throughout the myocardium, LGE often fails to identify or underestimates its extent due to the lack of normal myocardium as a background contrast. Secondly, LGE detects existing collagen deposits, meaning it can only identify existing scar tissue or focal necrosis. Essentially, it assesses static structural changes and cannot capture the real-time activation status of fibroblasts during the critical repair period after myocardial infarction.

[0006] In addition, CMR's T1 mapping and extracellular volume (ECV) techniques can also detect fibrosis, quantifying the expansion of the extracellular matrix and demonstrating strong detection capabilities for diffuse fibrosis. However, its signal is affected not only by collagen deposition but also by various factors such as myocardial edema, inflammation, and amyloid deposition. In the acute phase of acute myocardial infarction (AMI), due to severe myocardial edema, ECV is often overestimated and fails to accurately reflect the true fibrotic burden. Furthermore, CMR examination is expensive, time-consuming, and contraindicated in patients with implanted pacemakers, severe renal insufficiency, or claustrophobia.

[0007] [ 68Ga-DOTA-FAPI-04 PET / MR is a molecular imaging technique that utilizes a gallium-68-labeled fibroblast activation protein (FAP) inhibitor as a tracer. It achieves non-invasive visualization and quantitative monitoring of activated fibroblasts through the fusion of positron emission tomography (PET) and magnetic resonance imaging (MR). This technique primarily targets FAP, which is almost not expressed in quiescent fibroblasts, but highly expressed in activated myofibroblasts under pathological conditions (such as myocardial infarction, fibrosis, and cancer). Compared with other imaging techniques, [ 68 The advantages of Ga-DOTA-FAPI-04 PET / MR lie in its high specificity and quantitative capabilities, directly reflecting pathophysiological processes. However, due to the high cost of the equipment and the complexity of radionuclide preparation, [ 68 Ga]Ga-DOTA-FAPI-04 PET / MR is difficult to use as a routine screening method.

[0008] Given the current inadequacy of clinical quantification of myocardial fibrosis, several simple circulating biomarkers have been developed for assessing myocardial fibrosis activity. For example, galactoside-binding lectin 3 (Galectin-3) and soluble suppression of tumorigenicity 2 (sST2), while potentially related to fibrosis, exhibit complex dynamic changes during the acute phase of acute myocardial infarction (AMI), are significantly influenced by hemodynamics, and their specificity in predicting early fibroblast activation remains controversial. Furthermore, classic cardiac and inflammatory markers, such as troponin, natriuretic peptide, and C-reactive protein, lack specificity for myocardial fibrosis. Therefore, the development of a biomarker and diagnostic kit capable of early assessment of myocardial fibrosis activity in AMI patients is urgently needed.

[0009] HBP, also known as azurophilic granules or 37 kDa cationic antimicrobial protein (CAP37), is a secreted granule protein located in neutrophil secretory vesicles and azurophilic granules. The sequence of HBP is publicly available, for example, obtained with NCBI accession number NP001691 REGION: 27..248. HBP has been widely used to assess bacterial infections and sepsis. Recently, some researchers have also found that HBP can provide a prognostic risk warning for patients with ST-segment elevation myocardial infarction (STEMI), but whether it can be used to assess the degree of myocardial fibrosis activation remains unknown.

[0010] In summary, the existing technology lacks a simple, economical, non-invasive detection method that can specifically reflect the degree of myocardial fibrosis activation after AMI. Summary of the Invention

[0011] The purpose of this invention is to address the shortcomings of existing technologies by providing a plasma biomarker for early assessment of the activation level of myocardial fibroblasts in patients with acute myocardial infarction and its application; this application establishes for the first time the correlation between HBP and... [68 The quantitative correlation between Ga]Ga-DOTA-FAPI-04 uptake volume and cardiac structural and functional parameters was proposed and verified, thus proposing that HBP can be used to assess the degree of myocardial fibroblast activation. The circulating plasma HBP level in AMI patients within a specific time window was quantitatively detected as a non-invasive biological indicator reflecting the intensity of fibroblast activation in vivo.

[0012] To achieve the above objectives, this application adopts the following technical solution:

[0013] This application provides the use of a detection reagent in the preparation of a product for assessing or dynamically monitoring the activation level of myocardial fibroblasts in patients with acute myocardial infarction (AMI), said detection reagent comprising at least a reagent for detecting the level of heparin-binding protein (HBP) in the plasma of patients with acute myocardial infarction.

[0014] In some implementations, the test samples are plasma samples from patients with acute myocardial infarction upon admission and at 24, 48, and 72 hours after percutaneous coronary intervention (PCI).

[0015] In some embodiments, the detection reagent further includes at least one of the following reagents for detecting high-sensitivity cardiac troponin I, high-sensitivity C-reactive protein, and N-terminal B-type natriuretic peptide precursor.

[0016] In some embodiments, the product includes at least one of reagents, kits, test strips, chips, and detection devices.

[0017] In some embodiments, the product is a testing device, which includes a sample collection device, a sample testing device, and a diagnostic device; wherein: The sample collection device is configured to collect plasma samples from a subject who is a patient with acute myocardial infarction. The sample detection device is a device capable of detecting the level (content or concentration) of heparin-binding protein in the plasma sample; The diagnostic device includes a data acquisition module and a diagnostic module, wherein the data acquisition module is configured to... The diagnostic module is configured to acquire data detected by the sample detection device and to obtain data based on the data. The data obtained by the module can be used to evaluate or dynamically monitor the activation level of myocardial fibroblasts in patients with acute myocardial infarction.

[0018] In some embodiments, the acute myocardial infarction patient is a patient with ST-segment elevation myocardial infarction (STEMI).

[0019] Compared with the prior art, this application has the following beneficial effects: 1) The biomarker (HBP) proposed in this application can achieve in vivo non-invasive monitoring of the dynamic activity of myocardial fibrosis, making up for the inability of traditional imaging methods such as late gadolinium enhancement technology to capture the real-time activation status of fibroblasts during the critical repair period after myocardial infarction; by detecting the level of heparin-binding protein (HBP), the degree of myocardial fibrosis activation in STEMI patients can be effectively assessed at an early stage, ensuring that timely clinical intervention measures can be taken, and ultimately helping to improve the prognosis of such high-risk patients.

[0020] 2) The proposed approach significantly improves the predictive efficacy and incremental clinical value of early risk stratification: Traditional biomarkers (such as hs-cTnI, hs-CRP, and NT-proBNP) can reflect the level of myocardial injury or inflammation, but lack specificity for subsequent fibroblast activation and pathological remodeling. The statistical data of this application clearly show that introducing HBP detection into the conventional clinical prediction model can generate significant incremental predictive value: After adding HBP for 48 hours, the AUC for assessing FAPI UV significantly increased from 0.593 to 0.732 (P=0.002), and the net reclassification improvement index (NRI) reached 0.697 (P<0.001). This indicates that HBP can capture pathophysiological information that is directly related to fibrosis healing and cannot be covered by traditional indicators.

[0021] 3) The proposed method has superior safety and clinical applicability, and broadens the applicable population: The proposed method only requires the collection of peripheral venous blood, is non-invasive, radiation-free and has extremely low testing costs, and is especially suitable for high-risk patients with renal insufficiency, claustrophobia or implanted non-MRI compatible pacemakers, and has strong clinical applicability. Attached Figure Description

[0022] Figure 1 ROC curve for predicting UV elevation using plasma HBP levels 72 hours after PCI. Detailed Implementation

[0023] To make the technical solution of this application clearer and easier to understand, preferred embodiments are described in detail below with reference to the accompanying drawings.

[0024] Unless otherwise specified, the experimental or testing methods described in the following examples are conventional methods; the reagents and materials described are obtained from conventional commercial sources unless otherwise specified.

[0025] Example This embodiment provides the application of a reagent for detecting heparin-binding protein (HBP) in the preparation of diagnostic products (e.g., kits) for assessing the activation level of myocardial fibroblasts in AMI patients. 1. Study subjects and inclusion criteria This study was approved by the Ethics Committee of Ruijin Hospital, affiliated with Shanghai Jiao Tong University School of Medicine (2020-152), and informed consent was obtained from the participants. Participants included those diagnosed with STEMI at Ruijin Hospital, affiliated with Shanghai Jiao Tong University School of Medicine, between August 2023 and April 2024, and who completed the procedure 30±7 days after diagnosis. 68 Patients with Ga]Ga-DOTA-FAPI-04 PET / MR. A total of 74 patients were ultimately included in this study.

[0026] The selection criteria are as follows: Patients eligible to enroll in this study must meet all of the following criteria: (1) Written informed consent must be obtained before any assessment is conducted; (2) Male or female patients aged 18 or older; (3) Based on the generally accepted definition of myocardial infarction, the diagnosis is spontaneous AMI; Spontaneous acute myocardial infarction (AMI) is defined as clinical evidence of myocardial necrosis consistent with myocardial ischemia resulting from a major coronary event. In these cases, spontaneous AMI must be diagnosed according to the following criteria: An elevated and / or decreased cardiac enzyme (cardiac troponin, cTn, or CK-MB) was detected, with at least one value exceeding the 99th percentile of the upper limit of normal (URL) or the local laboratory's diagnostic threshold for MI, and at least one of the following pieces of evidence of myocardial ischemia was present: (1) Ischemic discomfort or other ischemic symptoms; (2) ECG features of STEMI, including new or presumed to be new significant ST-T changes; (3) The presence of newly appearing pathological Q waves or left bundle branch block on ECG (If the patient’s spontaneous MI is secondary to another medical condition, such as anemia, hypotension or arrhythmia, or if the spontaneous MI is thought to be caused by coronary artery spasm and the coronary arteries are normal, the patient is not eligible). Patients who presented with clinical manifestations associated with Takotsubo cardiomyopathy did not meet the inclusion criteria (the time of this AMI visit refers to the time when the patient went to the emergency room / emergency treatment center, was admitted to the cardiovascular intensive care unit or cardiology ward, etc., for the treatment of this AMI).

[0027] The exclusion criteria are as follows: Patients meeting any of the following criteria are not eligible for enrollment in this study: (1) Has a known history of chronic heart failure; (2) Had cardiogenic shock within 24 hours prior to enrollment; (3) Persistent clinical heart failure prior to enrollment; (4) Stroke or transient ischemic attack within one month prior to enrollment; (5) Before enrollment, the researchers assessed that the patient had serious infection, trauma, hematological diseases, surgery, or other diseases that might interfere with HBP measurement.

[0028] (6) For various reasons, cardiac MRI cannot be performed (eGFR<30, having a pacemaker or other metal implants that cannot be used for MRI, claustrophobia, etc.). (7) For various reasons, they are not suitable for radionuclide scanning (such as tracer allergy, presence of tumors affecting tracer uptake, etc.).

[0029] 2. Experimental Testing: Peripheral venous blood was collected from patients at admission and at 24, 48, and 72 hours after primary percutaneous coronary intervention (pPCI). Plasma samples were anticoagulated using a 1:9 sodium citrate solution. The samples were centrifuged at 3000 rpm for 15 minutes at 4°C. Strict avoidance of aspirating the leukocyte layer was required during plasma separation to prevent interference from high levels of hemoglobin (HBP) released by neutrophil degranulation. A dry fluorescence immunoassay was used with a Jet-iStar 3000 fully automated immunoassay analyzer (Zhonghan Shengteng Biotechnology Co., Ltd., Zhejiang, China). 50 µl of plasma sample was incubated for 18 minutes before measuring the HBP concentration (ng / mL).

[0030] 3. Statistical Analysis The effectiveness of this technical solution has been verified through rigorous statistical models. The statistical analysis related to this application includes the following: (1) Basic statistical description Normality was tested using the Kolmogorov-Smirnov test. Continuous variables that conform to a normal distribution are expressed as mean ± standard deviation (Mean ± SD); continuous variables that do not conform to a normal distribution are expressed as median (25th and 75th percentiles). Categorical variables are expressed as frequency (percentage).

[0031] (2) Correlation and intergroup analysis Spearman correlation analysis was used to assess the relationship between plasma HBP concentration and [ 68 Ga]Ga-DOTA-FAPI-04 PET / MR parameters, such as the correlation between FAPI uptake volume and cardiac structural and functional parameters.

[0032] The Wilcoxon rank-sum test was used to compare the differences in HBP levels among different fibrosis activity groups (stratified based on the median of FAPI PET parameters).

[0033] (3) Regression model Univariate and multivariate linear regression models were constructed, with HBP concentration as the primary exposure variable. After adjusting for clinical confounding factors, the independent predictive power of HBP on fibroblast activation volume and ventricular remodeling risk was determined by calculating standardized regression coefficients, 95% confidence intervals (CI), and p-values.

[0034] (4) Diagnostic incremental value assessment The diagnostic accuracy of HBP for highly activated fibroblasts was evaluated by constructing receiver operating characteristic (ROC) curves and calculating the area under the curve (AUC).

[0035] The Integrated Discriminant Improvement Index (IDI) and Net Reclassification Improvement Index (NRI) were calculated to quantitatively evaluate the incremental predictive contribution of adding HBP to traditional myocardial injury and inflammation indicators.

[0036] (5) P<0.05 is considered statistically significant.

[0037] (6) Use R (Bell Labs version 4.0.0) to analyze data and create graphs.

[0038] 4. Research Results (1) Result 1: Analysis of baseline clinical characteristics of subjects This prospective study included 74 STEMI patients who underwent emergency PCI. Baseline clinical characteristics are shown in Table 1. The median age of the participants was 65.00 years (IQR: 56.00, 71.00), with males accounting for 83.8% (62 cases), reflecting the typical demographic characteristics of STEMI. In terms of clinical characteristics, 94.6% of the patients presented with Killip Class I cardiac function, and the main culprit vessels were the left anterior descending artery (LAD, 39.2%) and the right coronary artery (RCA, 39.2%).

[0039] Patients' drug treatment followed standard guidelines, including dual antiplatelet therapy (98.6%), statins (91.9%), and ACEI / ARB / ARNI drugs (78.4%).

[0040] Table 1 Baseline clinical characteristics of enrolled patients

[0041] Note: Data are expressed as median (25th percentile, 75th percentile) or frequency (percentage).

[0042] Abbreviations: ACEI / ARB / ARNI, angiotensin-converting enzyme inhibitor / angiotensin II receptor blocker / angiotensin receptor neprilysin inhibitor.

[0043] (2) Result 2: [ 68 Ga]Ga-DOTA-FAPI-04 PET / MR characteristics Completed 30±7 days after STEMI [ 68 In Ga-DOTA-FAPI-04 PET / MR imaging, the subjects exhibited significant fibroblast activation and structural changes (see Table 2). The mean TBRmax of the subjects was 5.08 ± 1.61, the median TBRmean was 2.78, and the median FAPI uptake volume, representing the intensity of fibroblast activation, reached 98.28 cm³. 3In addition, the subjects' left ventricular ejection fraction (LVEF) was 49.78±9.95%, and the mean left ventricular end-systolic volume (LVESV) was 70.40±21.87 mL. The median infarct area (IS) accounted for 17.60% of the left ventricular myocardium, the median absolute IS volume was 20.90 mL, and the median area at risk accounted for 32.71%.

[0044] Table 2 [ 68 Ga]Ga-DOTA-FAPI-04 PET / MR results

[0045] Note: PET / MR: Positron Emission Tomography / Magnetic Resonance Imaging; TBR: Target-to-background ratio; UV: [ 68 Ga-FAPI-04 Uptake Volume; CMR: Cardiac Magnetic Resonance Imaging; LVM: Left Ventricular Mass; LVEDV / I: Left Ventricular End-Diastolic Volume / Index; LVVESV / I: Left Ventricular End-Symptom Volume / Index; LVEF: Left Ventricular Ejection Fraction; SV / I: Stroke Volume / Index; CO / CI: Cardiac Output / Cardiac Index; LVMI: Left Ventricular Mass Index; LGE: Late Gadolinium Enhancement; LVMM: Left Ventricular Myocardial Mass; IS: Infarct Area.

[0046] (2) Result 3: Plasma HBP level and [ 68 Correlation analysis of PET / M parameters of Ga-DOTA-FAPI-04 By performing Spearman correlation analysis on plasma HBP concentrations collected at different time points and imaging parameters 30±7 days post-surgery (see Table 3 for details), this invention reveals the key characteristics of HBP in assessing myocardial fibroblast activity and early remodeling.

[0047] Association between HBP and fibroblast activation intensity: Plasma HBP levels at 48 hours post-surgery were significantly positively correlated with TBRmax (r = 0.364, P = 0.001) and TBRmean (r = 0.256, P = 0.028), indicators reflecting fibroblast activation intensity. HBP levels at 72 hours post-surgery showed a significant positive correlation with FAPI uptake volume (UV), a measure of activation extent (r = 0.237, P = 0.042). These results indicate that the sustained increase in HBP during the acute phase is closely related to the degree of fibroblast activation in the myocardium during the later stages.

[0048] Association of HBP with Left Ventricular Remodeling and Cardiac Function: Data showed that 72 hours postoperatively, HBP was significantly positively correlated with left ventricular end-systolic volume (LVESV) (r = 0.234, P = 0.046) and left ventricular end-systolic volume index (LVESVI) (r = 0.236, P = 0.043); and significantly negatively correlated with left ventricular ejection fraction (LVEF) (r = -0.274, P = 0.019). Furthermore, 72 hours postoperatively, HBP also showed a significant negative association with cardiac output (CO) (r = -0.313, P = 0.007) and cardiac index (CI) (r = -0.307, P = 0.008), which reflect cardiac pumping efficiency.

[0049] Association between HBP and myocardial infarction area: At 72 hours post-operation, HBP was significantly positively correlated with the percentage of infarct area (r = 0.234, P = 0.044) and the absolute volume of infarct (r = 0.312, P = 0.007).

[0050] In conclusion, although HBP peaks upon admission, its levels 48 to 72 hours post-surgery have stronger biological relevance and clinical value for assessing in vivo fibroblast activation intensity and predicting adverse ventricular remodeling later.

[0051] Table 3 Plasma HBP levels and [ 68 Correlation analysis of Ga-DOTA-FAPI-04 PET / MR parameters

[0052] Note: Data are expressed using Spearman correlation coefficient (r) and corresponding p-values. This indicates that P < 0.05. This indicates P < 0.01. Abbreviations: TBRmax / mean: Maximum / mean target-to-background ratio. UV: [68Ga]Ga-FAPI-04 uptake volume. LGE: Late gadolinium enhancement. LVESV / I: Left ventricular end-systolic volume / index. LVEF: Left ventricular ejection fraction. IS: Infarct area.

[0053] (4) Result 4: Independent predictive value of plasma HBP for [68Ga]Ga-DOTA-FAPI-04 PET / MR parameters To further validate the predictive ability of acute-phase circulating hemoglobin (HBP) levels for myocardial fibroblast activation and ventricular remodeling, this study employed univariate and multivariate linear regression analyses (see Table 4 for details). The multivariate model corrected for key clinical factors such as age, sex, peak high-sensitivity C-reactive protein (hs-CRP), and peak high-sensitivity cardiac troponin I (hs-cTnI).

[0054] Prediction of fibroblast activation intensity: In both univariate and multivariate analyses after adjusting for confounding factors, HBP levels at 48 hours post-surgery were significantly and independently positively correlated with TBRmax, an indicator reflecting fibroblast activation intensity (adjusted beta = 0.014, 95% CI: 0.005, 0.023, P = 0.002). It was also an independent predictor of TBRmean (adjusted beta = 0.004, 95% CI: 0.000, 0.009, P = 0.032). Furthermore, HBP levels at 72 hours post-surgery were a key indicator for predicting fibroblast activation volume (UV). Multivariate regression analysis showed a significant and independent positive correlation between HBP at 72h and FAPI UV (adjusted beta = 0.571, 95% CI: 0.114, 1.028, P = 0.015).

[0055] Independent Prediction of Cardiac Structural Remodeling and Function: HBP at 72 hours post-procedure showed strong predictive value for cardiac remodeling. A adjusted multivariate model confirmed that HBP at 72 hours post-procedure was an independent predictor of increases in left ventricular end-systolic volume (LVESV) (beta = 0.233, P = 0.015) and left ventricular end-systolic volume index (LVESVI) (beta = 0.116, P = 0.026). Furthermore, HBP at 72 hours independently predicted a decrease in left ventricular ejection fraction (LVEF) (beta = -0.097, P = 0.036) and an increase in left ventricular mass (LVM) (beta = 0.139, P = 0.048).

[0056] Assessment of the extent of myocardial infarction: Regression analysis further confirmed that HBP at 72 hours postoperatively was significantly and independently positively correlated with the percentage of infarction assessed by magnetic resonance imaging (corrected beta = 0.117, P = 0.033) and the absolute volume of infarction (corrected beta = 0.098, P = 0.028).

[0057] The regression analysis results above clearly indicate that the circulating HBP level 48-72 hours after surgery is not only a reflection of myocardial injury or inflammation, but also a reliable and independent early biological warning indicator of the intensity of fibroblast activation and subsequent pathological ventricular remodeling.

[0058] Table 4. Plasma HBP on [ 68 Independent predictive value of Ga-DOTA-FAPI-04 PET / MR parameters

[0059] Statistical Explanation: The data in the table were analyzed using linear regression, and the regression coefficients beta, 95% confidence intervals (CI), and p-values ​​for univariate (unadjusted) and multivariate (adjusted) regressions are presented. The multivariate model has been corrected for age, sex, peak levels of high-sensitivity C-reactive protein (hs-CRP) and high-sensitivity cardiac troponin I (hs-cTnI). Abbreviations: HBP: Heparin-binding protein. PET / MR: Positron emission tomography / magnetic resonance imaging. TBRmax / mean: Target-to-background ratio. UV: [ 68 Ga]Ga-DOTA-FAPI-04 Uptake Volume. CMR: Cardiac Magnetic Resonance Imaging. LVM / I: Left ventricular mass / index. LVEDV / I: Left ventricular end-diastolic volume / index. LVESV / I: Left ventricular end-systolic volume / index. LVEF: Left ventricular ejection fraction. SV / I: Stroke volume / index. CO / CI: Cardiac output / cardiac index. IS: Infarct area. LVMM: Left ventricular myocardial mass.

[0060] (5) Result 5: Plasma HBP on [ 68 Predictive value and incremental diagnostic efficacy of Ga-DOTA-FAPI-04 uptake volume The effects of plasma HBP levels on in vivo fibroblast activation were systematically evaluated using ROC curve analysis, the Integrated Discriminant Improvement Index (IDI), and the Net Reclassification Improvement Index (NRI). 68 The predictive accuracy of Ga-DOTA-FAPI-04 uptake volume (based on the median UV) was investigated, and its incremental diagnostic value on top of existing clinical biomarkers was explored (see Table 5 and...). Figure 1 ).

[0061] Comparison of predictive efficacy of single biomarkers: This invention defines a UV level above the median as a highly activated state. Prediction of HBP levels at 72 hours post-surgery using a single biomarker.68 The area under the curve (AUC) of Ga-DOTA-FAPI-04 uptake volume UV reached 0.717, indicating that HBP 72 hours after pPCI has a strong discriminative efficacy against fibroblast activation. The optimal cutoff value was determined to be 42.07 ng / mL using the Youden's Index. At this cutoff value, the predictive sensitivity was 56.8%, and the specificity was as high as 83.8%. Figure 1 This result is significantly superior to traditional clinical biomarkers, including peak high-sensitivity cardiac troponin I (Peak hs-cTnI, AUC = 0.547), peak high-sensitivity C-reactive protein (Peak hs-CRP, AUC = 0.594), and peak N-terminal pro-B-type natriuretic peptide (Peak NT-proBNP, AUC = 0.527). This strongly demonstrates that plasma hemoglobin (HBP) can serve as an early warning indicator, specifically identifying high-risk patients with postoperative high-intensity myocardial fibrosis. An HBP level exceeding 42.07 ng / mL at 72 hours can serve as an important basis for clinical decision-making regarding the initiation of antifibrotic therapy or intensive follow-up.

[0062] Incremental value of HBP to conventional prediction models: Adding HBP for 72 hours to the Peak hs-cTnI base model significantly improved predictive performance (AUC increased from 0.547 to 0.723), with an IDI of 0.136 (P<0.001) and an NRI of 0.657 (P =0.004). Adding HBP for 72 hours to the Peak hs-CRP base model increased the model AUC to 0.730 and showed the best reclassification improvement ability (NRI = 0.737, P<0.001). A joint prediction model consisting of Peak hs-cTnI, Peak hs-CRP, and PeakNT-proBNP was established (basic AUC=0.593). After incorporating the HBP 72h index, the predictive performance of the integrated model was further improved to 0.732, with an IDI of 0.133 (P = 0.002) and an NRI of 0.697 (P = 0.002).

[0063] These results demonstrate that plasma hemoglobin (HBP) levels can capture fibroblast activation information that traditional injury, inflammation, and hemodynamic indicators cannot reflect. Introducing HBP detection into existing myocardial infarction risk assessment systems can significantly improve the accuracy of identifying active fibrosis formation in vivo, providing a strong incremental diagnostic basis for precise risk stratification of STEMI patients.

[0064] Table 5. Accuracy of cTnI, hs-CRP, and HBP in predicting uptake in STEMI patients.

[0065] Note: AUC: Area under the receiver operating characteristic curve. IDI: Overall discriminant improvement index. NRI: Net reclassification improvement index. Peak hs-cTnI: Peak value of high-sensitivity cardiac troponin I. Peak hs-CRP: Peak value of high-sensitivity C-reactive protein. Peak NT-proBNP: Peak value of N-terminal pro-B-type natriuretic peptide. HBP 48h: Plasma heparin-binding protein level 48 hours post-surgery. Ref: Reference model. NA: Not applicable.

[0066] The above description is merely a preferred embodiment of this application and is not intended to limit this application in any form or substance. It should be noted that those skilled in the art can make several improvements and additions without departing from this application, and these improvements and additions should also be considered within the scope of protection of this application.

Claims

1. The use of a detection reagent in the preparation of a product for assessing or dynamically monitoring the activation level of myocardial fibroblasts in patients with acute myocardial infarction, characterized in that, The test reagents include at least those for detecting the level of heparin-binding protein in the plasma of patients with acute myocardial infarction.

2. The application according to claim 1, characterized in that, The test samples were plasma samples from patients with acute myocardial infarction upon admission, or 24, 48, or 72 hours after percutaneous coronary intervention.

3. The application according to claim 1, characterized in that, The detection reagent also includes at least one of the following reagents for detecting high-sensitivity cardiac troponin I, high-sensitivity C-reactive protein, and N-terminal B-type natriuretic peptide precursor.

4. The application according to claim 1, characterized in that, The product includes at least one of reagents, kits, test strips, chips, and testing devices.

5. The application according to claim 4, characterized in that, The product is a testing device. The testing equipment includes a sample collection device, a sample testing device, and a diagnostic device; among which: The sample collection device is configured to collect plasma samples from a subject who is a patient with acute myocardial infarction. The sample detection device is a device capable of detecting the level of heparin-binding protein in the plasma sample; The diagnostic device includes a data acquisition module and a diagnostic module, wherein the data acquisition module is configured to... The diagnostic module is configured to acquire data detected by the sample detection device and to obtain data based on the data. The data obtained by the module can be used to evaluate or dynamically monitor the activation level of myocardial fibroblasts in patients with acute myocardial infarction.

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