Application of platelet-derived growth factor PDGF-D in assessment of right heart failure

Abstract

The invention discloses an application of a platelet-derived growth factor PDGF-D in evaluation of right heart failure. Specifically, the invention provides application of a substance for detecting PDGF-D in preparation of a product for right heart failure prognosis evaluation or right heart failure prognosis monitoring. The right heart failure is caused by pulmonary arterial hypertension. The introduction of PDGF-D can significantly improve the accuracy and sensitivity of right heart failure disease assessment, help clinicians to better understand the disease change of patients, and improve the effect of long-term treatment.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology and relates to the application of platelet-derived growth factor PDGF-D in the evaluation of right heart failure, and specifically relates to the use of PDGF-D as a new biomarker to specifically improve the accuracy of long-term treatment evaluation of clinical right heart failure. Background Art

[0002] Pulmonary hypertension (PH) is a serious cardiovascular disease that has brought a heavy health burden to the world. It is especially common in patients with underlying diseases such as chronic obstructive pulmonary disease and left heart disease.

[0003] In cases of right heart failure, pulmonary hypertension is one of the main causes. Studies have shown that among patients with pulmonary hypertension, 30% to 50% of cases eventually develop right heart failure (data from the Journal of the European Respiratory Society), especially in patients with other cardiopulmonary diseases, the incidence of right heart failure is higher.

[0004] The reason why pulmonary hypertension easily leads to right heart failure is that pulmonary vascular resistance increases and the afterload of the right ventricle also increases significantly. Long-term high load can lead to right ventricular remodeling, including myocardial fibrosis and right ventricular hypertrophy. In the early stage, the right ventricle can compensate through hypertrophy and dilation to maintain normal blood flow. However, this compensation can only relieve symptoms in the short term. As the disease progresses, the right ventricle gradually loses its compensatory ability, leading to right ventricular failure.

[0005] Currently, there are very limited reports on the role of PDGF-D (platelet derived growth factor D) in heart failure. Existing studies mainly focus on the role of PDGF-D in angiogenesis, but its specific application in right heart failure has not been fully studied and utilized.

[0006] Existing clinical detection methods, such as N-terminal pro-brain natriuretic peptide (NT-proBNP), have certain limitations in the assessment of right heart failure. Although NT-proBNP is widely used as a detection indicator for heart failure, it mainly reflects overall heart function problems and lacks specificity and sensitivity for right heart failure. This means that relying solely on NT-proBNP cannot fully reflect the state of right heart function. Therefore, there is an urgent need to develop new biomarkers to increase the evaluation efficiency of right heart failure and improve long-term clinical treatment effects. Summary of the invention

[0007] The present invention aims to solve the problem of the lack of specific and sensitive detection indicators for right heart failure in the prior art. The present invention, through in-depth research on platelet-derived growth factor D (PDGF-D), discovered its unique expression pattern and clinical detection value in patients with right heart failure, thereby providing a new, sensitive and specific biomarker, overcoming the shortcomings of the prior art, and providing strong support for the prognosis evaluation and personalized treatment of right heart failure in the clinic.

[0008] The experimental subjects of this study are right heart tissues from patients with clinical right heart failure, which are extremely rare and valuable. These samples are extremely difficult to obtain, providing the inventors with a hard-won research opportunity to deeply explore the potential application of PDGF-D in right heart failure.

[0009] The present invention solves the above technical problems through the following technical solutions.

[0010] On one hand, the present invention provides an application of a substance for detecting PDGF-D in the preparation of a product for evaluating the prognosis of right heart failure or monitoring the prognosis of right heart failure; the right heart failure is right heart failure caused by pulmonary hypertension.

[0011] In some embodiments, the substance for detecting PDGF-D is selected from one or more of the following:

[0012] 1) an antibody or antigen-binding fragment that specifically binds to PDGF-D;

[0013] 2) protein microarray; and,

[0014] 3) siRNA or shRNA targeting PDGF-D.

[0015] In some embodiments, the antibody or antigen-binding fragment that specifically binds to PDGF-D is, for example, Thermo Fisher PA5-114143 or H00080310-D01P.

[0016] In some embodiments, the antibody or antigen-binding fragment that specifically binds to PDGF-D can be made into a detection kit, such as a PDGF-D enzyme-linked immunosorbent assay (ELISA) kit (biotechwell, Cat #EH10901M).

[0017] In some embodiments, the substance for detecting PDGF-D is used to detect the molecular level of PDGF-D.

[0018] In some embodiments, the product includes a kit and a chip.

[0019] When the product for assessing right heart function or monitoring prognosis of right heart failure is a kit, it comprises an antibody or antigen-binding fragment that specifically binds to PDGF-D.

[0020] When the product for right heart function assessment or prognosis monitoring of right heart failure is a chip, microspheres of specific colors are used, each of which is bound to a different antibody or nucleic acid probe. The microspheres labeled with different antibodies are mixed with the sample. If the sample contains PDGF-D, it will bind to the antibody on the corresponding microsphere. A second fluorescent-labeled antibody is added, and the Luminex instrument is used to detect the fluorescent signal on the microsphere to determine the concentration of PDGF-D.

[0021] In some embodiments, the molecular level is selected from protein expression level or mRNA level.

[0022] Another aspect of the present invention provides a use of a substance for detecting PDGF-D in preparing a product for evaluating right heart function associated with pulmonary hypertension.

[0023] In some embodiments, the right heart function associated with pulmonary hypertension includes one or more of right ventricular end-diastolic volume, right ventricular end-systolic volume, right ventricular fractional area change, tricuspid annular systolic excursion, and right heart ejection fraction.

[0024] In some embodiments, the substance for detecting PDGF-D is used to detect the molecular level of PDGF-D.

[0025] In some embodiments, the molecular level is selected from protein expression level or mRNA level.

[0026] In some embodiments, the substance for detecting PDGF-D is selected from one or more of the following:

[0027] 1) an antibody or antigen-binding fragment that specifically binds to PDGF-D;

[0028] 2) protein microarray; and,

[0029] 3) siRNA or shRNA targeting PDGF-D.

[0030] In some embodiments, PDGF-D was compared with a series of clinical indicators such as plasma tests and cardiac catheterization tests in patients with clinically existing pulmonary hypertension and right heart failure, such as NT-proBNP. The P value of PDGF-D was 0.013, and the risk ratio was 1.002 (95% CI: 1.001-1.005), indicating that PDGF-D has statistically significant risk prediction in patients with pulmonary hypertension and right heart failure. In contrast, the P values ​​of other parameters such as mPAP (mean pulmonary artery pressure), mRAP (mean right atrial pressure), PVRi (pulmonary vascular resistance index) and NT-proBNP (N-terminal B-type natriuretic peptide precursor) did not reach the significant level.

[0031] At present, mPAP (mean pulmonary artery pressure), mRAP (mean right atrial pressure), PVRi (pulmonary vascular resistance index) and NT-proBNP are important markers for assessing the severity of pulmonary hypertension and right heart failure.

[0032] mPAP (mean pulmonary artery pressure) refers to the average pressure in the pulmonary artery during cardiac contraction and relaxation. Increased mPAP can lead to remodeling of pulmonary microvessels and cause pulmonary hypertension. Pulmonary hypertension leads to increased right ventricular afterload, which can cause right ventricular hypertrophy and dysfunction in the long term. Therefore, increased mPAP is an important indicator of pulmonary hypertension, reflecting the right ventricular load, and this indicator is usually measured by right heart catheterization.

[0033] mRAP (mean right atrial pressure) refers to the average pressure in the right atrium. When right heart failure occurs, the pumping function of the right ventricle decreases, causing blood to stagnate in the right atrium, thereby increasing mRAP. Measurement of mRAP helps to assess the preload of the right ventricle. It is an indirect indicator of right heart failure and can be assessed by cardiac ultrasound or Doppler echocardiography.

[0034] PVRi (pulmonary vascular resistance index) is a measure of pulmonary vascular resistance, which is the ratio of pulmonary artery pressure to cardiac output. In the case of right heart failure, pulmonary vascular resistance increases, which will lead to a further increase in right ventricular afterload and aggravate right heart failure. Therefore, an increase in PVRi is also an important indicator of pulmonary hypertension and right heart failure, which can be measured and calculated by right heart catheterization.

[0035] NT-proBNP (N-terminal pro-B-type natriuretic peptide) is a hormone released by ventricular myocytes when pressure and volume are overloaded. In right heart failure, ventricular wall tension increases, leading to increased secretion of NT-proBNP. NT-proBNP is a biomarker for assessing the severity of heart failure, and its level is positively correlated with the severity of heart failure. This indicator can be obtained through blood tests.

[0036] When PDGF-D is used as a clinical evaluation indicator, its long-term sensitivity can show higher specificity and sensitivity as the disease progresses over a long period of time.

[0037] Another aspect of the present invention provides an application of a substance for detecting PDGF-D in the screening and development of right heart failure drugs; the right heart failure is right heart failure caused by pulmonary hypertension.

[0038] In some embodiments, the substance for detecting PDGF-D is used to detect the molecular level of PDGF-D.

[0039] In some embodiments, the substance for detecting PDGF-D is used to evaluate the therapeutic effect of a test drug on right heart failure.

[0040] In some embodiments, the substance for detecting PDGF-D is used to identify the target site of the drug to be tested.

[0041] In some embodiments, the molecular level is selected from protein expression level or mRNA level.

[0042] In some embodiments, the substance for detecting PDGF-D is selected from one or more of the following:

[0043] 1) an antibody or antigen-binding fragment that specifically binds to PDGF-D;

[0044] 2) protein microarray; and,

[0045] 3) siRNA or shRNA targeting PDGF-D.

[0046] In some embodiments, substances that detect PDGF-D are used to assess and stratify right heart failure and to help determine the progression of right heart failure in a patient.

[0047] In addition, PDGF-D can also play a key role in efficacy evaluation and safety monitoring. It is used to evaluate the efficacy of drug treatment. For example, a decrease in PDGF-D levels after using the test drug may indicate that the test drug is effective in improving heart failure symptoms and slowing disease progression, thereby serving as an intermediate endpoint to evaluate the short-term effect of the drug.

[0048] For example, a new drug for the treatment of right heart failure is being developed. In the early stages of clinical trials, PDGF-D can be used as the primary endpoint to evaluate the initial efficacy of the drug. If the drug is found to significantly reduce PDGF-D levels and this change is associated with improvement in clinical symptoms, then the drug may continue to be tested in larger clinical trials.

[0049] Another aspect of the present invention provides a substance for detecting PDGF-D for use in the development of right heart failure-related biomarkers, or in the study of the pathological and physiological mechanism of right heart failure; the right heart failure is right heart failure caused by pulmonary hypertension.

[0050] In some embodiments, the substance for detecting PDGF-D is used to detect the molecular level of PDGF-D.

[0051] In some embodiments, the substance detecting PDGF-D is used to validate a newly discovered potential biomarker.

[0052] In some embodiments, the molecular level is selected from protein expression level or mRNA level.

[0053] In some embodiments, the substance for detecting PDGF-D is selected from one or more of the following:

[0054] 1) an antibody or antigen-binding fragment that specifically binds to PDGF-D;

[0055] 2) protein microarray; and,

[0056] 3) siRNA or shRNA targeting PDGF-D.

[0057] Therefore, the present invention shows that PDGF-D can be used as a benchmark to help develop new right heart failure-related markers. For example, it is known that mPAP, mRAP, PVRi and NT-proBNP are closely related to the pathophysiological mechanism of right heart failure. In the present invention, these indicators are used to verify that the PDGF-D of the present invention has a good correlation with right heart failure, and PDGF-D has the potential to be used as a biomarker.

[0058] Another aspect of the present invention provides a method for detecting PDGF-D and using the method as a biomarker in the preparation of a product having the following functions:

[0059] 1) Prognosis assessment of right heart failure; and / or,

[0060] 2) Right heart failure prognosis monitoring;

[0061] The right heart failure is right heart failure caused by pulmonary hypertension.

[0062] In some embodiments, the substance for detecting PDGF-D is used to detect the molecular level of PDGF-D.

[0063] In some embodiments, the molecular level is selected from protein expression level or mRNA level.

[0064] In some embodiments, the substance for detecting PDGF-D is selected from one or more of the following:

[0065] 1) an antibody or antigen-binding fragment that specifically binds to PDGF-D;

[0066] 2) protein microarray; and,

[0067] 3) siRNA or shRNA targeting PDGF-D.

[0068] On the basis of being in accordance with the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0069] The reagents and raw materials used in the present invention are commercially available.

[0070] The positive and progressive effects of the present invention are:

[0071] Enhanced prognosis assessment capabilities: Studies have shown that PDGF-D levels are closely related to patient prognosis. By monitoring PDGF-D, clinicians can be provided with a more comprehensive disease assessment tool to help optimize treatment strategies and improve patient prognosis.

[0072] Promote personalized treatment: As a new biomarker for right heart failure, PDGF-D will provide a scientific basis for the formulation of personalized treatment plans, enabling patients to receive more precise treatment and reduce hospitalization and mortality rates.

[0073] Support multidisciplinary collaboration: The clinical application of PDGF-D will promote interdisciplinary cooperation in cardiology, molecular biology and related disciplines, and promote the comprehensive development of heart failure research and clinical practice.

[0074] Discovery of new biomarkers: The first systematic study of the expression characteristics of PDGF-D in patients with right heart failure has opened up a new direction for the clinical detection of right heart failure.

[0075] Multi-level validation mechanism: This study verified the clinical application potential of PDGF-D through multiple approaches, including patient plasma samples, right heart tissues of rare right heart failure patients, animal models, and corresponding control sample tests and in vitro experiments, providing sufficient experimental data to support its rationality and effectiveness as a biomarker.

[0076] Highly targeted application value: PDGF-D has strong specificity and sensitivity in predicting clinical deterioration events. It can target the specific pathophysiological characteristics of right heart failure and significantly improve the applicability of existing detection methods in clinical practice.

[0077] Promoting clinical standardization: The implementation of the present invention is helpful to promote the standardized treatment of right heart failure, improve the overall level of clinical practice, and conform to the trend of modern medical development. BRIEF DESCRIPTION OF THE DRAWINGS

[0078] Figure 1 :Statistical analysis of the correlation between PDGF-D and NT-proBNP;

[0079] The figure shows that there is a significant positive correlation between PDGF-D and NT-proBNP levels, providing a theoretical basis for PDGF-D as a biomarker for evaluating right heart failure.

[0080] Figure 2 :Kaplan-Meier curves of patients with pulmonary hypertension and right heart failure based on PDGF-D levels;

[0081] The figure shows that the probability of no clinical worsening events in patients with low PDGF-D levels is significantly higher than that in patients with high PDGF-D levels, indicating a close correlation between PDGF-D levels and patient prognosis outcomes.

[0082] Figure 3 : Comparison of plasma PDGF-D concentrations before and after Potts surgery in patients with right heart failure and pulmonary hypertension, ** represents p value less than or equal to 0.01;

[0083] The figure shows that after patients with pulmonary hypertension underwent Potts surgery, right heart failure was significantly improved. The plasma PDGF-D concentration of these patients with pulmonary hypertension was lower after surgery than before surgery, and the difference was statistically significant, indicating that PDGF-D levels are closely correlated with patients' right heart function.

[0084] Figure 4 :COX proportional hazard model for prognostic evaluation of patients with pulmonary hypertension and right heart failure;

[0085] This figure shows that PDGF-D has statistically significant risk prediction in patients with pulmonary hypertension and right heart failure. In contrast, the P values ​​of other pulmonary hemodynamic parameters (mPAP, mRAP, PVRi) and right heart function parameters (NT-proBNP) did not reach the significant level.

[0086] Figure 5 : ROC curves of PDGF-D and NT-ProBNP as prognostic indicators;

[0087] This figure evaluates the performance of PDGF-D and NT-proBNP as prognostic indicators for right heart failure. Currently, NT-proBNP is a commonly used indicator for predicting right heart function in pulmonary hypertension in clinical practice. Due to the limited number of samples, the AUCs of NT-proBNP as a 1-year, 3-year, and 5-year prognostic marker in this study cohort were 0.5, 0.61, and 0.54, respectively. The 1-year, 3-year, and 5-year AUC values ​​of PDGF-D as a prognostic indicator for right heart failure were 0.64, 0.87, and 0.88, respectively. This suggests that when PDGF-D is used as a clinical prognostic monitoring indicator, its accuracy increases with the long-term development of the disease, and is higher than NT-proBNP. DETAILED DESCRIPTION

[0088] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0089] The embodiments of the present invention have discovered and verified the key role of PDGF-D in right heart remodeling in pulmonary hypertension through multi-level research.

[0090] In clinical samples, single-cell nuclear transcriptome sequencing was performed on right heart tissues of patients with pulmonary hypertension and controls, and it was found that the proportion of fibroblasts in the right heart of patients with pulmonary hypertension was significantly increased, and there were significant differences in cell proliferation ability. Further analysis revealed a subpopulation of fibroblasts with high proliferation activity, which specifically expressed PDGF-D. The significance of PDGF-D was verified between cell subpopulations and patient samples, suggesting that it may play an important role in the pathological development of right heart remodeling in pulmonary hypertension.

[0091] Establishment and validation of a mouse model of right heart failure

[0092] Model establishment: 8-week-old male C57BL / 6J mice (Beijing Vital River Laboratory Animal Technology Co., Ltd.) were placed in a hypoxia chamber (Shanghai Tower Intelligent Technology Co., Ltd.) and fed with hypoxia for 4 weeks (oxygen concentration was 10% O 2 ), simulating right heart failure caused by pulmonary hypertension.

[0093] Verification: Millar catheter was used to open the chest to detect the right ventricular systolic pressure of mice, confirming that the pulmonary hypertension model was successfully established. WGA staining and HE staining were then used to evaluate the right ventricular remodeling phenotype. Real-time quantitative polymerase chain reaction was used to detect the mRNA levels of Nppa and Nppb in right ventricular tissue, confirming that the right heart failure model was successfully established. The heart was then removed, the right ventricular tissue was separated, and it was quickly frozen in liquid nitrogen and stored at -80°C.

[0094] During the preparation of single cell nucleus suspension, the frozen right ventricular tissue was minced in a cell lysis buffer consisting of 0.1% NP40, 10 mM Tris-HCl, 146 mM NaCl, 1 mM CaCl 2 , 21 mM MgCl 2 After confirming that the cells were completely lysed by trypan blue staining, 1 mL ST Washbuffer (10 mM Tris-HCl, 146 mM NaCl, 1 mM CaCl 2 , 21 mM MgCl 2, 0.01% BSA (NEB B9000S) and 40 U / mL RNase inhibitor). Filter through a 40 µm cell sieve, transfer the filtrate to a 15 mL centrifuge tube, rinse the cell sieve with an appropriate amount of STWash buffer, and combine the rinse with the cell nucleus filtrate. Centrifuge at 500 g in a horizontal rotor at 4°C for 5 min. Resuspend the cell nuclei with 5 mL PBS (containing 1% BSA), wash and centrifuge, and resuspend the cell nuclei with 100 µL PBS (containing 1% BSA). Stain with trypan blue and count under a microscope.

[0095] The cell nuclei were diluted to a concentration of 1000 / µL. In the single-cell nuclear sequencing experiment, the machine and cDNA library amplification were performed according to the operating instructions of 10×GenomicsChromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 (1000268). The DNA library was constructed using the Chromium™ Single Cell 3' / 5' Library Construction Kit (1000020). The constructed library was sequenced on the Illumina sequencing platform using the PE150 sequencing mode. In the data analysis stage, the sequencing data were processed using Cell Ranger software and R packages (such as Seurat), including quality control, alignment, quantification, dimensionality reduction, and clustering analysis to identify specific genes that change during right heart failure, and compared with clinical sample cell marker genes and biological functions.

[0096] Similar results from human studies were observed in a mouse model of right heart failure with pulmonary hypertension: the proliferation of fibroblasts and the gene expression characteristics of specific subpopulations were closely related to PDGF-D. Further gene screening and enrichment analysis showed that PDGF-D also has significant expression and functional characteristics during the right heart remodeling process in mice.

[0097] Finally, in in vitro experiments, the promotion of PDGF-D on fibroblast proliferation, migration and cell cycle was verified by culturing mouse cardiac fibroblasts and treating them with recombinant PDGF-D protein (rmPDGF-DD). These experimental results further support the potential pathogenic mechanism of PDGF-D in right heart remodeling in pulmonary hypertension.

[0098] In summary, the core role of PDGF-D in right heart remodeling in pulmonary hypertension has been verified at multiple levels from clinical samples, animal models and cell experiments, and its potential as a prognostic monitoring indicator has been demonstrated.

[0099] Example 1 Statistical analysis of the correlation between PDGF-D and NT-proBNP

[0100] Venous blood samples were collected from 65 patients with pulmonary hypertension and right heart failure. PDGF-D concentration was determined by ELISA, and NT-proBNP concentration was determined by the laboratory using immunoassay and obtained through the clinical medical record system of Shanghai Children's Medical Center.

[0101] Sample preparation: 3 mL of venous blood was collected from patients with pulmonary hypertension and right heart failure, placed in an anticoagulant tube containing EDTA, and centrifuged at 3000 rpm for 10 minutes at 4°C. After separating the plasma, the sample was frozen and stored at -80°C for testing. When testing, the plasma sample was thawed and equilibrated at room temperature for 10 minutes.

[0102] PDGF-D concentration detection: PDGF-D enzyme-linked immunosorbent assay (ELISA) kit (biotechwell, Cat #EH10901M) was used to prepare the standard curve according to the kit instructions. After diluting the sample with 0.1M PBS, 100 μL of the sample was added to a 96-well ELISA plate and incubated at room temperature for 1 hour. After the incubation was completed, the reaction was stopped, the plate was washed and the secondary antibody was added, and incubated again at room temperature for 30 minutes. After washing the plate, the color developer was added to react, and finally the absorbance of the sample was read at a wavelength of 450 nm, and the concentration of PDGF-D was calculated using the standard curve.

[0103] All samples follow uniform sampling guidelines and are processed as described above to ensure data accuracy.

[0104] After data collection, correlation analysis was performed using SPSS or R statistical software. First, the normality of the data and the influence of outliers were confirmed, and the original data were transformed or corrected as necessary. Then, scatter plots were drawn using the "ggplot2" and "stats" packages in R software, and the Pearson correlation coefficient was used to calculate the correlation between PDGF-D and NT-proBNP. The correlation coefficient and confidence interval generated by R software were used to evaluate the efficacy of PDGF-D as a detection indicator. During data processing and analysis, all data points were ensured to be included in the analysis, and abnormal data and missing values ​​were removed.

[0105] like Figure 1 As shown in the results, there is a significant positive correlation between PDGF-D and the existing right heart failure detection indicator NT-proBNP, and this consistency is statistically significant (p=0.025). This lays a theoretical foundation for the clinical application of PDGF-D as a sensitive plasma detection indicator for patients with right heart failure. Therefore, PDGF-D is expected to become a new right heart failure assessment indicator with important clinical application value.

[0106] Example 2 Determination of Kaplan-Meier survival curve for patients with pulmonary arterial hypertension and right heart failure based on PDGF-D level

[0107] To evaluate the correlation between PDGF-D level and the survival rate of patients with pulmonary arterial hypertension and right heart failure, venous blood samples were collected from 55 patients with right heart failure, and the PDGF-D concentration in plasma was measured by ELISA according to the method of Example 1. The patients were divided into a low PDGF-D group of 27 cases and a high PDGF-D group of 28 cases according to the median PDGF-D concentration, and all samples were standardized before the experiment. Subsequently, clinical deterioration events of the patients, including death, heart transplantation, lung transplantation, and Potts surgery, were collected and long-term follow-up was conducted to record the time. Then, the Kaplan-Meier method was used for analysis to calculate the survival probability without clinical deterioration events of the patients in the low PDGF-D group and the high PDGF-D group. The Log-rank test was used to evaluate the survival difference between the two groups, and all data were processed in SPSS or R statistical software. The Kaplan-Meier survival curve was plotted in R software using the "survival" package, and the curve was calculated based on the survival data of each group. During the data analysis process, all patient data were standardized to ensure the accuracy of the results. When plotting the curve, appropriate confidence intervals and statistical tests were used for verification.

[0108] The inventor found in the clinical cohort that, as Figure 2 shown, according to the results of the Kaplan-Meier survival curve, the survival probability without clinical deterioration events of patients with low PDGF-D level was significantly higher than that of patients with high PDGF-D level, and there was a significant difference (p = 0.019). This indicates that the PDGF-D level is closely related to the long-term survival rate of patients, and a low PDGF-D level predicts a better prognosis. This also suggests that PDGF-D can be used as a potential clinical prognostic evaluation index to predict the survival rate of patients and help formulate personalized treatment plans.

[0109] Example 3 Comparison of plasma PDGF-D concentrations before and after Potts surgery in patients with pulmonary arterial hypertension and right heart failure

[0110] For 10 patients with pulmonary arterial hypertension and right heart failure who underwent Potts surgery clinically, venous blood samples were collected before and after the surgery, and the PDGF-D concentration was measured by ELISA according to the method shown in Example 1. The experimental results showed that the clinical symptoms of the patients with right heart failure were improved after Potts surgery. As Figure 3 shown, the plasma PDGF-D concentration decreased significantly after the surgery, suggesting the potential of PDGF-D as a detection index in the clinical prognosis of patients with right heart failure.

[0111] Example 4 COX hazard ratio model for prognosis assessment of patients with pulmonary hypertension and right heart failure

[0112] A total of 49 patients with pulmonary hypertension and right heart failure were collected, and the risk of PDGF-D was compared with the existing plasma markers of pulmonary hypertension and right heart failure, NT-proBNP, and cardiac catheterization detection indicators. mPAP (mean pulmonary artery pressure) and mRAP (mean right atrial pressure) were obtained through clinical right heart catheterization. The pulmonary vascular resistance index (PVRi) was calculated based on mPAP and cardiac output. Cardiac output was calculated based on pediatric oxygen consumption, aortic oxygen saturation, superior vena cava oxygen saturation, and hemoglobin level. The data sources were obtained through the clinical medical record system of Shanghai Children's Medical Center.

[0113] Figure 4 The results showed that the P value of PDGF-D was 0.013 and the hazard ratio was 1.002 (95% CI: 1.001-1.005), indicating that PDGF-D has significant risk prediction in patients with right heart failure due to pulmonary hypertension. In contrast, the P values ​​of other parameters such as mPAP, mRAP, PVRi and NT-proBNP did not reach the significant level. These results support the clinical application potential of PDGF-D as a new indicator for the prognosis assessment of right heart failure and prove its advantage in assessment accuracy.

[0114] Example 5 ROC curves of PDGF-D and NT-ProBNP as prognostic indicators

[0115] In order to compare the predictive ability of PDGF-D and NT-proBNP at different time points, long-term follow-up data were collected for 49 patients. The PDGF-D concentration was determined by ELISA according to the method of Example 1, and the NT-proBNP concentration and related clinical indicators were obtained through the clinical medical record system. Curve drawing and data analysis were completed by R software, and the ROC curve was generated using the "pROC" package. According to the PDGF-D concentration and NT-proBNP concentration at different time points and the patient's clinical deterioration events, the curve was generated and the corresponding AUC value was calculated. The red line in the curve represents the AUC for 1 year, the green line represents the AUC for 3 years, and the blue line represents the AUC for 5 years. The dotted line is the baseline for random prediction (AUC = 0.5). The closer the curve is to the upper left corner, the higher the prediction accuracy of the model. The final data was standardized to ensure the accuracy of the curve generation.

[0116] The inventors used PDGF-D and NT-proBNP as prognostic evaluation indicators for right heart failure, and evaluated their prognostic prediction performance using ROC curves. Figure 5As shown in the figure, the AUC values ​​of NT-proBNP as a 1-year, 3-year and 5-year prognostic marker were 0.5, 0.61 and 0.54, respectively. The AUC values ​​of PDGF-D as a prognostic indicator of right heart failure were 0.64, 0.87 and 0.88, respectively, at 1 year, 3 years and 5 years. This indicates that when PDGF-D is used as a clinical prognostic predictor, its prediction accuracy increases with the progression of the disease, and it can show higher specificity and sensitivity, which is higher than NT-proBNP. This result shows that PDGF-D has very reliable clinical application value and prospects in the long-term treatment evaluation and monitoring of patients with right heart failure.

Claims

1. Use of a substance for detecting PDGF-D in the preparation of a product for right heart failure prognosis assessment or right heart failure prognosis monitoring; the right heart failure is right heart failure caused by pulmonary hypertension.

2. The use according to claim 1, characterized in that The substance for detecting PDGF-D is selected from one or more of the following: 1) an antibody or antigen-binding fragment that specifically binds to PDGF-D; 2) protein microarray; and, 3) siRNA or shRNA targeting PDGF-D; And / or, the substance for detecting PDGF-D is used to detect the molecular level of PDGF-D; And / or, the product includes a kit and a chip; Preferably, the molecular level is selected from protein expression level or mRNA level.

3. Use of a substance for detecting PDGF-D in the preparation of a product for evaluating right heart function associated with pulmonary hypertension; Preferably, the substance for detecting PDGF-D is used to detect the molecular level of PDGF-D; More preferably, the molecular level is selected from protein expression level or mRNA level.

4. The use according to claim 3, characterized in that The substance for detecting PDGF-D is selected from one or more of the following: 1) an antibody or antigen-binding fragment that specifically binds to PDGF-D; 2) protein microarray; and, 3) siRNA or shRNA targeting PDGF-D.

5. Use of a substance for detecting PDGF-D in the screening and development of drugs for right heart failure; the right heart failure is right heart failure caused by pulmonary hypertension; Preferably, the substance for detecting PDGF-D is used to detect the molecular level of PDGF-D; And / or, the substance for detecting PDGF-D is used to evaluate the therapeutic effect of the drug to be tested on right heart failure; And / or, the substance for detecting PDGF-D is used to identify the target of the drug to be tested; More preferably, the molecular level is selected from protein expression level or mRNA level.

6. The use according to claim 5, characterized in that The substance for detecting PDGF-D is selected from one or more of the following: 1) an antibody or antigen-binding fragment that specifically binds to PDGF-D; 2) protein microarray; and, 3) siRNA or shRNA targeting PDGF-D.

7. A substance for detecting PDGF-D is used in the development of biomarkers related to right heart failure, or in the study of the pathophysiological mechanism of right heart failure; the right heart failure is right heart failure caused by pulmonary hypertension; Preferably, the substance for detecting PDGF-D is used to detect the molecular level of PDGF-D; And / or, the substance for detecting PDGF-D is used to verify a newly discovered potential biomarker; More preferably, the molecular level is selected from protein expression level or mRNA level.

8. The use according to claim 7, characterized in that The substance for detecting PDGF-D is selected from one or more of the following: 1) an antibody or antigen-binding fragment that specifically binds to PDGF-D; 2) protein microarray; and, 3) siRNA or shRNA targeting PDGF-D.

9. Use of a substance for detecting PDGF-D as a biomarker in the preparation of a product having the following functions: 1) Prognosis assessment of right heart failure; and / or, 2) Right heart failure prognosis monitoring; The right heart failure is right heart failure caused by pulmonary hypertension; Preferably, the substance for detecting PDGF-D is used to detect the molecular level of PDGF-D; More preferably, the molecular level is selected from protein expression level or mRNA level.

10. The use according to claim 9, characterized in that The substance for detecting PDGF-D is selected from one or more of the following: 1) an antibody or antigen-binding fragment that specifically binds to PDGF-D; 2) protein microarray; and, 3) siRNA or shRNA targeting PDGF-D.

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