Metabolic markers for assessing surgical outcome in patients with hypertrophic cardiomyopathy and applications

Abstract

The application belongs to the technical field of molecular biology, and particularly relates to metabolic markers for evaluating the prognosis of patients with hypertrophic cardiomyopathy and application thereof. The application determines that one or more of 3-hydroxyrilotene, asparagine-lysine, cyclohexylamine, arachidonic acid, aconitic acid, propionyl carnitine, ceramide, tryptophan, 3-hydroxysebacic acid, traumatic acid, phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine and S-garlicin cysteine are related to the prognosis of hypertrophic cardiomyopathy, can evaluate the prognosis of patients with hypertrophic cardiomyopathy after undergoing muscle resection of interventricular septum, and shows accurate judgment on all-cause death or rehospitalization, cardiovascular events and postoperative new-onset arrhythmia. Finally, all the prognostic biomarkers are reproduced in an independent cohort, which is helpful for postoperative risk evaluation of patients with hypertrophic cardiomyopathy, has high specificity and high sensitivity.

Description

Technical Field

[0001] This invention belongs to the field of molecular biology technology, specifically relating to metabolic biomarkers and their applications in evaluating the surgical prognosis of patients with hypertrophic cardiomyopathy. Background Technology

[0002] Hypertrophic cardiomyopathy (HCM) is a cardiovascular disease characterized by myocardial hypertrophy, clinically manifested as increased thickness of the left ventricular wall. As one of the most common inherited cardiovascular diseases, HCM has a prevalence of 1 in 200–500 in the general population, affecting the health of 20 million people worldwide. HCM has become a significant cause of sudden cardiac death, heart failure, and atrial fibrillation, placing a tremendous burden on modern healthcare systems and the economy.

[0003] Currently, there are no reliable clinical indicators for predicting the prognosis of HCM patients, and potential clinical indicators related to HCM prognosis have not demonstrated accurate predictive ability. Given the limitations of current HCM diagnostic methods and the lack of prognostic indicators for HCM patients, this field seeks new indicators and methods for accurately diagnosing HCM and precisely predicting the prognosis of HCM patients. Summary of the Invention

[0004] The purpose of this invention is to provide metabolic biomarkers and their applications for assessing the surgical prognosis of patients with hypertrophic cardiomyopathy (HCM), to diagnose, predict, assess, and stratify adverse events after HCM surgery, to predict the risk of death after surgery, and to inform patients to take preventive and treatment measures in advance.

[0005] This invention provides biomarkers for assessing the prognosis of hypertrophic cardiomyopathy, including one or more of a first biomarker, a second biomarker, and a third biomarker;

[0006] The first biomarker includes one or more of 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconic acid;

[0007] The second biomarker includes one or more of propionylcarnitine, ceramide, tryptophan, 3-hydroxysepiacetic acid, and traumatic acid;

[0008] The third biomarker includes one or more of phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-allicin cysteine.

[0009] The present invention also provides the application of the biomarkers described in the above technical solution in the preparation of products for assessing the prognosis of hypertrophic cardiomyopathy.

[0010] Preferably, the prognosis includes the prognosis of adverse postoperative events; the adverse postoperative events include one or more of all-cause death or readmission, cardiovascular events, and new-onset arrhythmias.

[0011] The present invention also provides the application of the biomarkers described in the above technical solution in the preparation of products for diagnosing adverse events after surgery for hypertrophic cardiomyopathy; the adverse events after surgery include one or more of all-cause death or readmission, cardiovascular events, and new-onset arrhythmias.

[0012] The present invention also provides the application of the biomarkers described in the above technical solution in the preparation of products for risk assessment and / or risk stratification of adverse events after surgery for hypertrophic cardiomyopathy; the adverse events after surgery include one or more of all-cause death or readmission, cardiovascular events and new-onset arrhythmias.

[0013] Preferably, the hypertrophic cardiomyopathy includes hypertrophic obstructive cardiomyopathy.

[0014] Preferably, the product includes a reagent kit.

[0015] The present invention also provides a kit for diagnosis, prognosis, risk assessment and risk stratification of all-cause mortality or readmission after surgery for hypertrophic cardiomyopathy, including reagents for detecting a first biomarker;

[0016] The first biomarker includes one or more of 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconic acid.

[0017] The present invention also provides a kit for diagnosing, prognosticating, assessing risk and stratifying risk of cardiovascular events after surgery for hypertrophic cardiomyopathy, including a reagent for detecting a second biomarker;

[0018] The second biomarker includes one or more of propionylcarnitine, ceramide, tryptophan, 3-hydroxysepiacetic acid, and traumatic acid.

[0019] The present invention also provides a kit for the diagnosis, prognosis, risk assessment and risk stratification of new-onset arrhythmias after surgery for hypertrophic cardiomyopathy, including reagents for detecting a third biomarker;

[0020] The third biomarker includes one or more of phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-allicin cysteine.

[0021] Beneficial effects:

[0022] This invention establishes a biomarker panel to predict adverse postoperative outcomes in patients with hypertrophic cardiomyopathy by analyzing the relationship between metabolic and clinical characteristics. It identifies plasma metabolites associated with several comorbidities in patients with hypertrophic cardiomyopathy and reveals systemic metabolic disorders caused by complex comorbidities. A group of prognostic biomarkers for hypertrophic cardiomyopathy (HCM) were identified, including one or more of the following: a first biomarker, a second biomarker, and a third biomarker. The first biomarker included one or more of 3-methylhistidine, asparagine-lysine, and cyclohexylamine. The second biomarker included one or more of 3-hydroxybutyrylcarnitine, acetylmethionine, 3-hydroxydecanoic acid, asparagine-glutamine, L-ergothioneine, and traumatic acid. The third biomarker included one or more of PC (16:0 / 18:2), PC (18:1 / 16:0), creatine, L-carnitine, and hemolytic PC (22:4). These biomarkers were able to assess the prognosis of HCM patients undergoing ventricular septal resection, demonstrating accurate prediction of all-cause mortality or readmission, cardiovascular events, and new postoperative arrhythmias. Finally, all prognostic biomarkers were reproducible in an independent cohort, aiding in postoperative risk assessment for HCM patients with high specificity and sensitivity. Attached Figure Description

[0023] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the embodiments will be briefly described below.

[0024] Figure 1 To derive the ROC curve analysis results of individual biomarkers screened by LASSO regression between the two groups in the cohort;

[0025] Figure 2 The ROC curve analysis results of the prognostic model constructed using the binary LR algorithm for the queue are shown in the figure.

[0026] Figure 3 The Kaplan-Meier curve (log-rank test) analysis results for deriving adverse events in cohort HCM patients;

[0027] Figure 4 To verify the ROC curve analysis results of individual biomarkers screened by LASSO regression between the two groups in the cohort;

[0028] Figure 5 The ROC curve analysis results of the prognostic model constructed using the binary LR algorithm for the queue are shown in the figure.

[0029] Figure 6 The Kaplan-Meier curve (log-rank test) analysis results of adverse events in the cohort of HCM patients were used to verify the results. Detailed Implementation

[0030] This invention provides biomarkers for assessing the prognosis of hypertrophic cardiomyopathy, including one or more of a first biomarker, a second biomarker, and a third biomarker;

[0031] The first biomarker includes one or more of 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconic acid;

[0032] The second biomarker includes one or more of propionylcarnitine, ceramide, tryptophan, 3-hydroxysepiacetic acid, and traumatic acid;

[0033] The third biomarker includes one or more of phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-allicin cysteine.

[0034] In one embodiment, the 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconic acid are respectively present in plasma. The 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconic acid described in this invention are associated with all-cause mortality or readmission in patients with hypertrophic cardiomyopathy after ventricular septal resection. This allows for the assessment of the prognosis of hypertrophic cardiomyopathy patients after ventricular septal resection, demonstrating accurate judgment of all-cause mortality or readmission with high specificity and sensitivity.

[0035] In one embodiment, propionylcarnitine, ceramide, tryptophan, 3-hydroxydecanoic acid, and traumatic acid are respectively present in plasma as propionylcarnitine, ceramide, tryptophan, 3-hydroxydecanoic acid, and traumatic acid. The propionylcarnitine, ceramide, tryptophan, 3-hydroxydecanoic acid, and traumatic acid described in this invention are associated with cardiovascular events after ventricular septal resection in patients with hypertrophic cardiomyopathy, enabling the assessment of the prognosis after ventricular septal resection in patients with hypertrophic cardiomyopathy, demonstrating accurate judgment of cardiovascular events with high specificity and sensitivity.

[0036] In one embodiment, the phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-allicin cysteine ​​are respectively the plasma phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-allicin cysteine. The phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-allicin cysteine ​​described in this invention are respectively associated with new-onset arrhythmias after ventricular septal resection in patients with hypertrophic cardiomyopathy, enabling the assessment of the prognosis of patients with hypertrophic cardiomyopathy after ventricular septal resection, demonstrating accurate judgment of new-onset arrhythmias with high specificity and sensitivity.

[0037] Therefore, the application of the biomarker in the preparation of products for assessing the prognosis of hypertrophic cardiomyopathy also falls within the scope of protection of this invention. As one embodiment, the prognosis includes postoperative adverse event prognosis; the postoperative adverse events include one or more of all-cause death or readmission, cardiovascular events, and new-onset arrhythmias.

[0038] Therefore, the application of the biomarker in the preparation of products for diagnosing adverse events after surgery for hypertrophic cardiomyopathy; wherein the adverse events include one or more of all-cause death or readmission, cardiovascular events and new-onset arrhythmias, are also within the scope of protection of this invention.

[0039] Therefore, the application of the biomarker in the preparation of products for risk assessment and / or risk stratification of adverse events after surgery for hypertrophic cardiomyopathy; the adverse events after surgery, including one or more of all-cause death or readmission, cardiovascular events and new-onset arrhythmias, are also within the scope of protection of this invention.

[0040] In one embodiment, the hypertrophic cardiomyopathy includes hypertrophic obstructive cardiomyopathy. In another embodiment, the product includes a kit.

[0041] This invention also provides a kit for the diagnosis, prognosis, risk assessment, and risk stratification of all-cause mortality or readmission after surgery for hypertrophic cardiomyopathy, including reagents for detecting a first biomarker. The first biomarker includes one or more of 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconic acid.

[0042] This invention does not have strict requirements on the types of other components in the kit; conventional selection is acceptable. Using the kit described in this invention, only the levels of one or more of 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconic acid in the subject's plasma need to be detected. No gene testing is required to accurately assess the prognosis of patients with hypertrophic cardiomyopathy after ventricular septal resection, and to precisely determine all-cause mortality or readmission. The kit exhibits high specificity and sensitivity.

[0043] The present invention also provides a kit for the diagnosis, prognosis, risk assessment and risk stratification of cardiovascular events after surgery for hypertrophic cardiomyopathy, including a reagent for detecting a second biomarker; the second biomarker includes one or more of propionylcarnitine, ceramide, tryptophan, 3-hydroxysepiacetic acid and traumatic acid.

[0044] This invention does not have strict requirements on the types of other components in the kit; conventional selection is acceptable. Using the kit described in this invention, only the levels of one or more of propionylcarnitine, ceramide, tryptophan, 3-hydroxysepiacetic acid, and traumatic acid in the subject's plasma need to be detected. No gene testing is required to accurately assess the prognosis of patients with hypertrophic cardiomyopathy after ventricular septal resection, and to precisely determine cardiovascular events. The kit exhibits high specificity and sensitivity.

[0045] The present invention also provides a kit for the diagnosis, prognosis, risk assessment and risk stratification of new-onset arrhythmias after surgery for hypertrophic cardiomyopathy, including a reagent for detecting a third biomarker; said third biomarker includes one or more of phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine and S-allicin cysteine.

[0046] This invention does not have strict requirements on the types of other components in the kit; conventional selection is acceptable. Using the kit described in this invention, only one or more of the following—phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-allicin cysteine—need to be detected in the subject's plasma. No gene testing is required to accurately assess the prognosis of patients with hypertrophic cardiomyopathy after ventricular septal resection, and to precisely identify new-onset arrhythmias. The kit exhibits high specificity and sensitivity.

[0047] To further illustrate the present invention, the metabolic biomarkers and their applications for evaluating the surgical prognosis of patients with hypertrophic cardiomyopathy provided by the present invention are described in detail below with reference to the accompanying drawings and embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0048] Example 1

[0049] Sample collection

[0050] The inventors' team collected data from 441 patients with obstructive hypertrophic cardiomyopathy (HCM) at Fuwai Hospital, Chinese Academy of Medical Sciences. The diagnostic criteria for obstructive HCM were based on the 2024 ACC / AHA Guidelines for Primary Prevention of Cardiovascular Disease and the 2023 ESC Guidelines, as follows: echocardiography showing a maximum left ventricular wall thickness ≥15 mm. Patients who had previously undergone septal myotomy and alcohol septal ablation were excluded. Clinical examination and echocardiography showed no cardiac disease in all normal controls. To minimize metabolite degradation, fasting blood samples were processed into frozen plasma within 1 hour. Whole blood was centrifuged at 4000 rpm for 10 minutes at 4°C, and the plasma supernatant was collected and aliquoted into 0.2 mL Eppendorf tubes (70 μL plasma per tube). The plasma was immediately quenched in liquid nitrogen and stored at -80°C until analysis. This study was approved by the Ethics Committee of Fuwai Hospital and conducted in accordance with the Declaration of Helsinki, 1964. Written informed consent was provided by all individuals participating in this study. The clinical characteristics of the subjects are shown in Table 1.

[0051] Table 1 Clinical characteristics of the subjects

[0052]

[0053] Note: Continuous variables are represented by the median (25th-75th percentile).

[0054] Example 2

[0055] Patient follow-up

[0056] The follow-up period began with the first examination after surgery (septal myotomy) and ended on the last contact date or at death. All patients were followed up at least once a year via outpatient visit or telephone interview. The primary endpoint was all-cause mortality or readmission, and cardiovascular events (cardiac death, life-threatening arrhythmias, myocardial infarction, heart failure, and stroke). The secondary endpoint was new-onset arrhythmias postoperatively. Arrhythmic events were collected and categorized into three groups: (1) atrioventricular block; (2) atrial arrhythmias, including atrial fibrillation, atrial flutter, or paroxysmal atrial tachycardia (AT); and (3) ventricular arrhythmias, including symptomatic non-sustained ventricular tachycardia (VT), sustained VT, or ventricular fibrillation.

[0057] Example 3

[0058] Relationship between metabolic and clinical characteristics in HCM patients

[0059] (1) Clinical characteristics of HCM patients, especially the diagnosis and management of comorbidities, are crucial for the precise treatment of HCM. Based on postoperative follow-up data of HCM patients, the inventors used univariate and multivariate logistic regression analysis to assess the relationship between plasma metabolites and clinical characteristics of HCM. Atrial fibrillation (AF) is a common complication of HCM, and the inventors found a positive correlation between phosphocholine and AF. This correlation suggests that phosphocholine can be used to predict the risk of AF in patients, indicating the need for more frequent electrocardiographic monitoring and more aggressive management strategies, including consideration of intraoperative intervention. Similarly, myocardial bridging is another important comorbidity of HCM. Several metabolites, such as Glu-Ser and glucose, have shown correlation with myocardial bridging. These metabolites can be used to assess myocardial perfusion and provide guidance for coronary artery bypass grafting in the presence of myocardial bridging. Provocation testing in potential HCM patients can improve the detection rate of occult HCM. Plasma metabolites, including serine, can be used in conjunction with echocardiography to help diagnose occult HCM and reduce the risk of missed diagnosis in HCM patients. In addition, metabolites can be used to assess septal morphology in HCM patients, such as basal hypertrophy, midventricular hypertrophy, and apical hypertrophy. Metabolic markers also have predictive value for cardiac structural damage and diastolic dysfunction.

[0060] (2) Metabolic markers can serve as powerful tools for preoperative management and diagnostic characterization of HCM patients. Heart failure is a critical issue in the management of cardiomyopathy patients, and metabolic markers, including citrate, homocysteine, and leucine, can be used to assess the tendency for heart failure in HCM patients. Metabolic markers can also reflect the degree of left ventricular outflow tract obstruction in HCM patients, such as 2-hexenocarnitine. Severe left ventricular outflow tract obstruction is not solely caused by septal hypertrophy; it often involves the left ventricle, especially the posterior wall. The posterior wall of the left ventricle is adjacent to the membranous septum, through which conduction bundles pass, making it a key area for arrhythmia intervention. Posterior wall hypertrophy can lead to residual obstruction after septal myotomy. Assessing cardiac morphology based on metabolic markers can provide patients with more surgical insights and help in developing more precise surgical strategies. Metabolites can also provide evidence of the patient's overall condition, aiding in preoperative assessment of blood glucose levels and renal function.

[0061] (3) In addition, the inventors compared the metabolic characteristics of obstructive HCM patients with different genetic backgrounds. Principal component analysis showed that these characteristics were similar among patients with sarcomere variants, non-sarcomere variants, variants of indeterminate significance, and no variants. In summary, metabolic markers provide a comprehensive tool for preoperative decision-making, helping to answer what should be done and how to perform the surgery.

[0062] Example 4

[0063] A panel of biomarkers for predicting adverse postoperative outcomes in HCM patients was established using LASSO regression. Then, key metabolites that differentiated HCM patients from adverse postoperative events were selected using the LASSO regression algorithm, including all-cause mortality or readmission, cardiovascular events, and new postoperative arrhythmias. From the minimum bias model with optimal lambda, five metabolites were associated with all-cause death or readmission: 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconitate; five metabolites were associated with cardiovascular events: propionyl carnitine, ceramide (SM d18:0 / 22:3), tryptophan, 3-hydroxysebacic acid, and traumatic acid; and five metabolites were associated with new-onset postoperative arrhythmias: phosphatidylcholine (PC) 16:0 / 18:2), alanine-proline (Ala-pro), lysophosphatidylcholine (LysoPC 22:4), taurine, and S-allylcysteine.

[0064] Given the limited predictive power of single biomarkers for adverse outcomes, the inventors developed a biomarker panel based on these metabolites using a binary LR algorithm for ROC curve analysis to predict the risk of adverse events after ventricular septal resection in HCM patients. Results showed good predictive performance in both the derivation and validation cohorts. Figures 1-6 (and Tables 2-3).

[0065] Table 2. ROC analysis results of the derivation of the cohort prognostic model

[0066]

[0067]

[0068] Table 3. ROC analysis results of the validation cohort prognostic model

[0069]

[0070] As can be seen from the above, the biomarkers provided by this invention have potential translational application value and can be used for risk stratification and personalized treatment decisions for HCM patients in clinical practice.

[0071] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. The application of biomarkers in the preparation of products for assessing the prognosis of hypertrophic cardiomyopathy; wherein the prognosis is the prognosis of postoperative adverse events; wherein the postoperative adverse events are all-cause death or readmission; The biomarkers are 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconic acid; The hypertrophic cardiomyopathy includes hypertrophic obstructive cardiomyopathy.

2. The application according to claim 1, characterized in that, The product includes a reagent kit.

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