Metabolic marker for evaluating operation prognosis of hypertrophic cardiomyopathy patient and application

Abstract

The invention belongs to the technical field of molecular biology, and particularly relates to a metabolic marker for evaluating operation prognosis of a hypertrophic cardiomyopathy patient and application. The preparation method comprises the following steps: determining 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, aconic acid, propionyl carnitine, ceramide, tryptophan, 3-hydroxysebacic acid, traumatic acid, phosphatidylcholine, alanine-proline and lysophosphatidylcholine; one or more of taurine and S-allicin cysteine are related to the prognosis of hypertrophic cardiomyopathy, so that the prognosis of a hypertrophic cardiomyopathy patient after ventricular septal muscle resection can be evaluated, and accurate judgment on total cause death or rehospitalization, cardiovascular events and postoperative new arrhythmia is shown. And finally, all prognosis biomarkers are reproduced in an independent queue, so that postoperative risk assessment of hypertrophic cardiomyopathy patients is facilitated, the specificity is high, and the sensitivity is high.

Description

Technical Field

[0001] The present invention belongs to the technical field of molecular biology, and particularly relates to metabolic markers for evaluating the surgical prognosis of patients with hypertrophic cardiomyopathy and their applications. Background Art

[0002] Hypertrophic cardiomyopathy (HCM) is a cardiovascular disease characterized by myocardial hypertrophy, and its clinical features are manifested as an increase in the thickness of the left ventricular wall. As one of the most common hereditary cardiovascular diseases, the prevalence of HCM in the general population is as high as 1:200 - 500, affecting the health of 20 million people globally. HCM has become an important cause of sudden cardiac death, heart failure, and atrial fibrillation, imposing a huge burden on the modern social healthcare system and economy.

[0003] Currently, there is no clinical indicator that can well predict the prognosis of HCM patients, and potential clinical indicators related to the prognosis of HCM patients have not shown precise predictive ability. Given the limitations of current HCM diagnostic methods and the lack of indicator signs for the prognosis of HCM patients, the field expects new indicators and new methods for accurately diagnosing HCM and precisely predicting the prognosis of HCM patients. Summary of the Invention

[0004] The purpose of the present invention is to provide metabolic markers for evaluating the surgical prognosis of patients with hypertrophic cardiomyopathy and their applications, diagnose, prognose, assess risks, and perform risk stratification on adverse events after hypertrophic cardiomyopathy surgery, achieve prediction of the death risk of HCM patients after surgery, and inform patients to take preventive and treatment measures in advance.

[0005] The present invention provides markers for evaluating the prognosis of hypertrophic cardiomyopathy, including one or more of a first marker, a second marker, and a third marker;

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

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

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

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

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

[0011] The present invention also provides the use of the biomarker described in the above technical solution in the preparation of a product for diagnosing postoperative adverse events of hypertrophic cardiomyopathy; the postoperative adverse events include one or more of all-cause death or readmission, cardiovascular events, and new-onset arrhythmias.

[0012] The present invention also provides the use of the biomarker described in the above technical solution in the preparation of a product for risk assessment and / or risk stratification of postoperative adverse events of hypertrophic cardiomyopathy; the postoperative adverse events 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 kit.

[0015] The present invention also provides a kit for diagnosing, prognosticating, risk-assessing, and risk-stratifying all-cause death or readmission after surgery for hypertrophic cardiomyopathy, comprising a reagent for detecting a first biomarker;

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

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

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

[0019] The present invention also provides a kit for diagnosing, prognosticating, risk-assessing, and risk-stratifying new-onset arrhythmias after surgery for hypertrophic cardiomyopathy, comprising a reagent for detecting a third biomarker;

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

[0021] Beneficial effects:

[0022] By analyzing the relationship between the metabolic characteristics and clinical characteristics of patients with hypertrophic cardiomyopathy, the present invention establishes a biomarker panel for predicting adverse postoperative outcomes in patients with hypertrophic cardiomyopathy, identifies plasma metabolites associated with several comorbidities in patients with hypertrophic cardiomyopathy, and reveals the systemic metabolic disorders caused by complex comorbidities. A group of biomarkers related to the prognosis of hypertrophic cardiomyopathy is screened, including one or more of a first biomarker, a second biomarker, and a third biomarker; the first biomarker includes one or more of 3-methylhistidine, asparagine-lysine, and cyclohexylamine; the second biomarker includes one or more of 3-hydroxybutyrylcarnitine, acetylmethionine, 3-hydroxydecanoic acid, asparagine-glutamine, L-ergothioneine, and traumatic acid; the third biomarker includes one or more of PC(16:0 / 18:2), PC(18:1 / 16:0), creatine, L-carnitine, and lysophosphatidylcholine(22:4), which can evaluate the prognosis of patients with hypertrophic cardiomyopathy after ventricular septal myectomy, showing accurate judgment of all-cause death or readmission, cardiovascular events, and new-onset postoperative arrhythmias. Finally, all prognostic biomarkers are reproduced in an independent cohort, which helps in the postoperative risk assessment of patients with hypertrophic cardiomyopathy, with high specificity and sensitivity. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments.

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

[0025] Figure 2 Results of ROC curve analysis of the prognostic model constructed using the binary LR algorithm in the derivation cohort;

[0026] Figure 3 Results of Kaplan-Meier curve (log-rank test) analysis of adverse events in HCM patients in the derivation cohort;

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

[0028] Figure 5 Results of ROC curve analysis of the prognostic model constructed using the binary LR algorithm in the validation cohort;

[0029] Figure 6 Results of Kaplan-Meier curve (log-rank test) analysis of adverse events in HCM patients in the validation cohort. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0030] The present invention provides markers for evaluating the prognosis of hypertrophic cardiomyopathy, including one or more of a first marker, a second marker, and a third marker;

[0031] The first marker includes one or more of 3-hydroxy lidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconitic acid;

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

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

[0034] As an embodiment, the 3-hydroxy lidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconitic acid are respectively 3-hydroxy lidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconitic acid in plasma. The 3-hydroxy lidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconitic acid of the present invention are respectively related to all-cause death or readmission after septal myectomy in patients with hypertrophic cardiomyopathy, and can evaluate the prognosis of patients with hypertrophic cardiomyopathy after septal myectomy, showing accurate judgment of all-cause death or readmission, with strong specificity and high sensitivity.

[0035] As an embodiment, the propionylcarnitine, ceramide, tryptophan, 3-hydroxysebacic acid, and traumatic acid are respectively propionylcarnitine, ceramide, tryptophan, 3-hydroxysebacic acid, and traumatic acid in plasma. The propionylcarnitine, ceramide, tryptophan, 3-hydroxysebacic acid, and traumatic acid of the present invention are respectively related to cardiovascular events after septal myectomy in patients with hypertrophic cardiomyopathy, and can evaluate the prognosis of patients with hypertrophic cardiomyopathy after septal myectomy, showing accurate judgment of cardiovascular events, with strong specificity and high sensitivity.

[0036] As an embodiment, the phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-allylcysteine are respectively phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-allylcysteine in plasma. The phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-allylcysteine of the present invention are respectively related to new-onset arrhythmia after septal myectomy in patients with hypertrophic cardiomyopathy, and can evaluate the prognosis of patients with hypertrophic cardiomyopathy after septal myectomy, showing accurate judgment of new-onset arrhythmia, with strong specificity and high sensitivity.

[0037] In view of this, the application of the biomarker in the preparation of a product for evaluating the prognosis of hypertrophic cardiomyopathy also falls within the protection scope of the present invention. As an implementation manner, the prognosis includes the prognosis of adverse events after surgery; the adverse events after surgery include one or more of all-cause death or readmission, cardiovascular events, and new-onset arrhythmia.

[0038] In view of this, the application of the biomarker in the preparation of a product 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 arrhythmia also falls within the protection scope of the present invention.

[0039] In view of this, the application of the biomarker in the preparation of a product 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 arrhythmia also falls within the protection scope of the present invention.

[0040] As an implementation manner, the hypertrophic cardiomyopathy includes hypertrophic obstructive cardiomyopathy. As an implementation manner, the product includes a kit.

[0041] The present invention also provides a kit for diagnosing, prognosticating, risk assessing, and risk stratifying all-cause death or readmission after surgery for hypertrophic cardiomyopathy, including reagents for detecting a first biomarker. The first biomarker includes one or more of 3-hydroxy lidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconitic acid.

[0042] The present invention does not have strict requirements for the types of other components in the kit, and conventional selection can be made. By using the kit of the present invention, it is only necessary to detect the levels of one or more of 3-hydroxy lidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid, and aconitic acid in the plasma of a subject, and gene detection is not required, so as to accurately evaluate the prognosis of a patient with hypertrophic cardiomyopathy after ventricular septal myectomy, accurately judge all-cause death or readmission, with strong specificity and high sensitivity.

[0043] The present invention also provides a kit for diagnosing, prognosticating, risk assessing, and risk stratifying cardiovascular events after surgery for hypertrophic cardiomyopathy, including reagents for detecting a second biomarker; the second biomarker includes one or more of propionyl carnitine, ceramide, tryptophan, 3-hydroxysebacic acid, and traumatic acid.

[0044] The present invention has no strict requirements for the types of other components in the kit, and conventional selection can be made. By using the kit of the present invention, it is only necessary to detect the level of one or more of propionylcarnitine, ceramide, tryptophan, 3-hydroxysebacic acid, and traumatic acid in the plasma of the subject. Without gene detection, the prognosis of patients with hypertrophic cardiomyopathy after ventricular septal myectomy can be accurately evaluated, and cardiovascular events can be accurately judged, with strong specificity and high sensitivity.

[0045] The present invention also provides a kit for diagnosing, prognosticating, risk assessing, and risk stratifying new-onset arrhythmia after surgery for hypertrophic cardiomyopathy, including reagents for detecting a third biomarker; the third biomarker includes one or more of phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-sulfinylcysteine.

[0046] The present invention has no strict requirements for the types of other components in the kit, and conventional selection can be made. By using the kit of the present invention, it is only necessary to detect the level of one or more of phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine, and S-sulfinylcysteine in the plasma of the subject. Without gene detection, the prognosis of patients with hypertrophic cardiomyopathy after ventricular septal myectomy can be accurately evaluated, and new-onset arrhythmia can be accurately judged, with strong specificity and high sensitivity.

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

[0048] Example 1

[0049] Sample collection

[0050] The inventor team collected 441 patients with obstructive hypertrophic cardiomyopathy (HCM) from Fuwai Hospital, Chinese Academy of Medical Sciences. The diagnostic criteria for obstructive HCM were based on the 2024 "ACC / AHA Primary Prevention of Cardiovascular Disease Guidelines" and the 2023 "ESC Guidelines" as follows: Echocardiography showed a maximum left ventricular wall thickness ≥ 15 mm. Patients who had previously undergone septal myectomy and alcohol septal ablation were excluded. Clinical examinations and echocardiography showed that all normal controls had no heart diseases. 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 invention research was approved by the Fuwai Hospital Ethics Committee and conducted in accordance with the Declaration of Helsinki of 1964. Individuals participating in this study provided written informed consent. 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 expressed as median (25th - 75th percentile).

[0054] Example 2

[0055] Patient follow-up

[0056] The follow-up period started from the first examination after surgery (septal myectomy) and ended at the last contact date or death. All patients were followed up at least once a year through outpatient visits or telephone interviews. The primary endpoints were all-cause death or readmission, cardiovascular events (cardiac death, life-threatening arrhythmias, myocardial infarction, heart failure, and stroke). The secondary endpoint was new-onset arrhythmia after surgery. Arrhythmia events were collected and classified into three categories: (1) atrioventricular block; (2) atrial arrhythmias, including atrial fibrillation, atrial flutter, or paroxysmal atrial tachycardia (AT); (3) ventricular arrhythmias, including symptomatic non-sustained ventricular tachycardia (VT), sustained VT, or ventricular fibrillation.

[0057] Example 3

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

[0059] (1) The clinical characteristics of patients with HCM, especially comorbid diagnoses and management, are crucial for the precise treatment of HCM. Based on the postoperative follow-up data of HCM patients, the inventors used univariate and multivariate logistic regression analyses to evaluate the relationship between plasma metabolites and the clinical characteristics of HCM. Atrial fibrillation (AF) is a common complication of HCM, and the inventors found that phosphocholine was positively correlated with AF. This correlation indicates that phosphocholine can be used to predict the risk of AF in patients, indicating the need for more frequent electrocardiogram monitoring and more aggressive management strategies, including considering intraoperative intervention. Similarly, myocardial bridging is another important comorbidity of HCM. Several metabolites, such as Glu-Ser and glucose, showed correlations with myocardial bridging. These metabolites can be used to evaluate myocardial perfusion and provide guidance for coronary artery bypass grafting in the case of combined myocardial bridging. Performing a provocation test on potential HCM patients can increase the detection rate of occult HCM. 22 Plasma metabolites, including serine, can be used in combination with echocardiography to assist in the diagnosis of occult HCM and reduce the risk of missed diagnosis in HCM patients. In addition, metabolites can be used to evaluate the septal morphology of 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 the preoperative management and diagnostic characteristics of HCM patients. Heart failure is a key issue in the management of cardiomyopathy patients, and metabolic markers can be used to evaluate the tendency of heart failure in HCM patients, including citric acid, homocysteine, and leucine. Metabolic markers can also reflect the degree of left ventricular outflow tract obstruction in HCM patients, such as 2-hexenoyl carnitine. Severe left ventricular outflow tract obstruction is not caused solely by septal hypertrophy; it usually involves the left ventricle, especially the posterior wall. The posterior wall of the left ventricle is adjacent to the membranous septum, and the conduction bundle passes through this area, so it is a key area for arrhythmia intervention. Posterior wall hypertrophy may lead to residual obstruction after septal myectomy. Evaluating the cardiac morphology based on metabolic markers can provide more surgical insights for patients and help provide more precise surgical strategies. Metabolites can also provide evidence of the patient's overall condition and help evaluate blood glucose levels and renal function preoperatively.

[0061] (3) In addition, the inventors also 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 uncertain 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 biomarker panel for predicting postoperative adverse outcomes in HCM patients was established using LASSO regression. Then, the LASSO regression algorithm was used to select key metabolites that distinguish HCM patients with postoperative adverse events, including all-cause death or readmission, cardiovascular events, and new-onset postoperative arrhythmia. From the minimum deviation model with the optimal lambda, five metabolites were associated with all-cause death or readmission, including 3-Hydroxylidocaine, Asp-lys, Cyclohexylamine, Arachidic acid, and Aconitate; five metabolites were associated with cardiovascular events, including Propionyl carnitine, SM d18:0 / 22:3, Tryptophan, 3-Hydroxysebacic acid, and Traumatic Acid; five metabolites were associated with new-onset postoperative arrhythmia, including PC 16:0 / 18:2, Ala-pro, LysoPC 22:4, Taurine, and S-Allylcysteine.

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

[0065] Table 2 Results of ROC analysis of the prognostic model in the derivation cohort

[0066]

[0067]

[0068] Table 3 Results of ROC analysis of the prognostic model in the validation cohort

[0069]

[0070] As can be seen from the above, the biomarker provided by the present invention has potential translational application value and can be used for risk stratification and personalized treatment decision-making of HCM patients in clinical practice.

[0071] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, rather than all embodiments. People can also obtain other embodiments based on these embodiments without creative efforts, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A marker for evaluating the prognosis of hypertrophic cardiomyopathy, characterized in that: including one or more of a first marker, a second marker, and a third marker; The first marker includes one or more of 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid and aconitic acid; The second marker includes one or more of propionylcarnitine, ceramide, tryptophan, 3-hydroxysebacic acid and traumatic acid; The third marker includes one or more of phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine and S-allicin cysteine.

2. Use of the marker according to claim 1 in the preparation of a product for evaluating the prognosis of hypertrophic cardiomyopathy.

3. The use according to claim 2, characterized in that: The prognosis includes the prognosis of adverse events after surgery; the adverse events after surgery include one or more of all-cause death or re-admission, cardiovascular events and new arrhythmias.

4. Use of the marker of claim 1 in the preparation of a product for diagnosing adverse events after surgery for hypertrophic cardiomyopathy; the adverse events after surgery include one or more of all-cause mortality or re-admission, cardiovascular events, and new-onset arrhythmias.

5. Use of the marker described in claim 1 in the preparation of a product 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 mortality or re-hospitalization, cardiovascular events and new-onset arrhythmias.

6. The use according to any one of claims 2 to 5, characterized in that: The hypertrophic cardiomyopathy includes hypertrophic obstructive cardiomyopathy.

7. The use according to any one of claims 2 to 5, characterized in that: The products include kits.

8. A kit for diagnosing, prognosing, assessing and stratifying all-cause mortality or readmission after surgery for hypertrophic cardiomyopathy, characterized in that: including a reagent for detecting a first marker; The first marker includes one or more of 3-hydroxylidocaine, asparagine-lysine, cyclohexylamine, arachidonic acid and aconitic acid.

9. A kit for diagnosing, prognosing, risk assessing and risk stratifying cardiovascular events after surgery for hypertrophic cardiomyopathy, characterized in that: comprising a reagent for detecting a second marker; The second marker comprises one or more of propionylcarnitine, ceramide, tryptophan, 3-hydroxysebacic acid and traumatic acid.

10. A kit for diagnosing, prognosing, risk assessing and risk stratifying new-onset arrhythmias after surgery for hypertrophic cardiomyopathy, characterized in that: including a reagent for detecting a third marker; The third marker includes one or more of phosphatidylcholine, alanine-proline, lysophosphatidylcholine, taurine and S-allicin cysteine.

Images