Application of pyruvate kinase M2 in the diagnosis and prognosis of heart failure

Abstract

The present disclosure relates to the use of pyruvate kinase M2 in the diagnosis and prognosis of heart failure, belonging to the field of in vitro diagnostic technology. After excluding patients with malignant tumors, the present disclosure found that the level of pyruvate kinase M2 in the plasma of patients with heart failure was higher than that of normal patients without heart failure. In particular, the level in patients with left ventricular failure and total heart failure was significantly higher than that of normal patients without heart failure. The level in the plasma of patients with acute heart failure was also significantly higher than that of normal patients without heart failure. As a marker for heart failure, PKM2 has high accuracy in diagnosing heart failure and assessing its prognosis, providing a new and improved option for the prediction, diagnosis, auxiliary diagnosis, and prognosis of heart failure.

Description

Technical Field

[0001] The present disclosure relates to methods for diagnosing and predicting heart failure. Furthermore, the present disclosure relates to methods for diagnosing and prognosticating heart failure using pyruvate kinase M2. Background Art

[0002] Heart failure (HF) is a syndrome characterized by impaired ventricular filling and / or ejection function due to various structural or functional cardiac diseases. This results in an inability of cardiac output to meet the metabolic needs of the body's tissues, leading to pulmonary and / or systemic congestion and inadequate organ and tissue blood perfusion. Clinical manifestations include dyspnea and fluid retention (pulmonary congestion and peripheral edema) caused by venous blood stasis and inadequate arterial perfusion.

[0003] HF is the terminal stage of various cardiovascular diseases and a major cause of death from cardiovascular disease. It is a major public health issue. Accurate diagnosis of HF facilitates proactive and effective treatment, reduces cardiovascular events, and thus alleviates the burden on society and families. The diagnosis of HF includes clinical symptoms and signs, imaging examinations such as two-dimensional echocardiography and cardiac magnetic resonance imaging, and HF-related blood markers. Detection of HF-related blood markers is a crucial tool for diagnosing and assessing clinical prognosis in HF patients. Commonly used HF markers include brain natriuretic peptide (BNP), N-terminal pro-BNP (NT-proBNP), troponin T / I (cTnT / I), human galectin-3 (Gal-3), soluble ST-2, GDF-15, H-FABP, and CRP. Levels of these blood markers reflect the pathophysiological changes at various stages of HF. They are associated with changes in pressure load, inflammatory responses, myocardial injury, myocardial remodeling, and myocardial fibrosis during the HF process. They can diagnose and guide HF treatment at various levels, and contribute to acute risk stratification and prognosis in HF. Circulating BNP and NT-proBNP are the most widely used biomarkers in clinical practice, with their elevation positively correlated with the severity of HF. However, in certain conditions, such as lung diseases, neuroendocrine disorders, multiple organ failure (renal failure, liver failure), and severe infectious diseases, circulating BNP and NT-proBNP can also be significantly elevated, interfering with the clinical diagnosis of HF. Therefore, single biomarker testing often fails to accurately determine the severity and prognosis of HF. Therefore, combined testing of multiple biomarkers is often used in clinical diagnosis and guidance for HF.

[0004] Pyruvate kinase (PK) is a key, rate-limiting enzyme in the glycolytic pathway, catalyzing the conversion of phosphoenolpyruvate to pyruvate. It plays a central role in cellular energy metabolism. Four isoenzymes (PKM1, PKM2, PKR, and PKL) have been identified in mammals. The distribution of PKMs is tissue-specific. Under physiological conditions, PKM1 is expressed in most adult tissues, while PKM2 is primarily expressed during embryonic development. PKM2 consists of four subunits and exists as monomers, dimers, and tetramers. Extensive evidence indicates that PKM2 expression is elevated in various tumor tissues. PKM2 promotes high glucose uptake by tumor cells, inhibits oxidative phosphorylation, and generates large amounts of lactate, providing a favorable environment for tumor cell growth. Recent studies have linked PKM2 to activation of the glycolytic pathway during myocardial reperfusion injury and myocardial ischemia-hypoxia, as well as to the cardiomyocyte cell cycle and oxidative stress.

[0005] Biochem Biophys Res Commun. 2015 April 10; 459(3): 430-436, discloses that PKM2 expression is significantly increased in cardiac tissue of patients with heart failure, but does not study the expression of PKM2 in blood or plasma. US20070238781A1 discloses an in vitro method for detecting cardiac insufficiency or chronic heart failure, which comprises comparing the level of pyruvate kinase type M2 in a patient's plasma sample with a predetermined standard level, wherein the level of pyruvate kinase type M2 in the plasma sample is detected by an immunological assay using a monoclonal antibody to detect M2 pyruvate kinase. Summary of the Invention

[0006] In one aspect, the present disclosure provides a method for predicting, diagnosing, assisting in the diagnosis or prognosis of heart failure by detecting the level of PKM2 or mRNA encoding PKM2 protein in a subject, comprising detecting the level of PKM2 or mRNA encoding PKM2 protein in a sample from the subject, and comparing it with a predetermined standard level of PKM2 or mRNA encoding PKM2 protein, wherein a detectable increase relative to the predetermined standard level indicates the possibility of heart failure.

[0007] On the other hand, the present disclosure also provides the use of a reagent for detecting PKM2 or mRNA encoding PKM2 protein in the preparation of a kit for predicting, diagnosing, assisting in the diagnosis or prognostic evaluation of heart failure. The reagent is used to detect the level of PKM2 or mRNA encoding PKM2 protein in a subject sample and compare it with a predetermined standard level of PKM2 or mRNA encoding PKM2 protein. A detectable increase in the level of PKM2 or mRNA encoding PKM2 protein in the subject sample relative to the predetermined standard level indicates the possibility of heart failure.

[0008] In some embodiments, the reagent comprises an antibody or antigen-binding fragment thereof that specifically binds to PKM2. In some embodiments, the reagent comprises a reagent that specifically detects mRNA encoding the PKM2 protein. In some embodiments, the reagent comprises specific primers for amplifying mRNA encoding the PKM2 protein, or a specific probe or chip for detecting mRNA encoding the PKM2 protein.

[0009] Another aspect of the present disclosure provides the use of a reagent for detecting PKM2 or mRNA encoding PKM2 protein in the preparation of a kit for prognostic assessment of a patient with heart failure, wherein the patient is receiving anti-heart failure treatment, the prognostic assessment comprising: a. measuring the level of PKM2 or mRNA encoding PKM2 protein in at least two biological samples obtained from the patient at different times, wherein the different times are a first time point and a second time point, wherein the second time point is later than the first time point, and wherein the patient is being treated for heart failure; and b. comparing the levels of PKM2 or mRNA encoding PKM2 protein in the at least two biological samples, wherein a decrease in the level of PKM2 or mRNA encoding PKM2 protein collected at the second time point compared to the first time point indicates that the treatment is effective, and wherein an increase in the level of PKM2 or mRNA encoding PKM2 protein collected at the second time point compared to the first time point indicates that the treatment is not effective.

[0010] Another aspect of the present disclosure provides the use of a reagent for detecting PKM2 or mRNA encoding PKM2 protein in the preparation of a kit for diagnosing or assisting in the diagnosis of heart failure, the diagnosis comprising: a. measuring the level of PKM2 or mRNA encoding PKM2 protein in a sample obtained from a subject presenting symptoms of heart failure; and b. comparing with a predetermined standard level of PKM2 or mRNA encoding PKM2 protein, wherein a detectable increase in the level of PKM2 or mRNA encoding PKM2 protein in the sample relative to the predetermined standard level indicates the possibility of heart failure.

[0011] In some embodiments, the heart failure symptom is selected from dyspnea, cough, expectoration, hemoptysis, fatigue, dizziness, palpitations, decreased exercise capacity, oliguria, symptoms of renal impairment, cold extremities, forced sitting, confusion, cyanosis of the lips, pale complexion, sweating, irritability, pulmonary edema, lower extremity edema, hepatosplenomegaly, ascites, chest tightness, and shortness of breath. In some embodiments, the heart failure symptom is dyspnea. In some embodiments, the dyspnea is acute dyspnea.

[0012] The present disclosure also relates to the use of a reagent for detecting PKM2 or mRNA encoding PKM2 protein in the preparation of a kit for prognostic assessment of heart failure patients who are receiving anti-heart failure treatment, wherein the prognostic assessment includes:

[0013] a. measuring the level of PKM2 or mRNA encoding PKM2 protein in a patient sample upon admission; and

[0014] b. comparing the level of PKM2 or mRNA encoding PKM2 protein in the patient at the time of admission to the hospital with a reference value.

[0015] In some embodiments, if the patient's PKM2 level is higher than 50% of the reference value, the patient has a poor prognosis; if the patient's PKM2 level is lower than 50% of the reference value, the patient has a better prognosis. In some embodiments, the poor prognosis means that the incidence of death within one year is higher than 20%, higher than 25%, or higher than 30%. In some embodiments, the better prognosis means that the incidence of death within one year is lower than 15%, lower than 10%, or lower than 8%. In some embodiments, the better prognosis means that the incidence of cardiovascular death and heart failure readmission within one year is lower than 10%; the poor prognosis means that the incidence of cardiovascular death and heart failure readmission within one year is higher than 30%.

[0016] In some embodiments, the reference value is the average level of PKM2 or mRNA encoding PKM2 protein in a population of heart failure patients. In some embodiments, the reference value of PKM2 is selected from 15 to 25 U / L. In some embodiments, the reference value of PKM2 is selected from 15 to 20 U / L. In some embodiments, the reference value of PKM2 is selected from 18 to 20 U / L. In some embodiments, the reference value of PKM2 is selected from 18.0 U / L, 18.1 U / L, 18.2 U / L, 18.3 U / L, 18.4 U / L, 18.5 U / L, 18.6 U / L, 18.7 U / L, 18.8 U / L, 18.9 U / L, 19.0 U / L, 19.1 U / L, 19.2 U / L, 19.3 U / L, 19.4 U / L, 19.5 U / L, 19.6 U / L, 19.7 U / L, 19.8 U / L, 19.9 U / L or 20.0 U / L.

[0017] In some embodiments, the heart failure is left heart failure, global heart failure, acute heart failure, chronic heart failure, heart failure caused by ischemic myocardial damage, heart failure caused by hypertension, heart failure caused by alcoholic cardiomyopathy, heart failure caused by dilated cardiomyopathy, heart failure caused by hypertrophic cardiomyopathy, heart failure caused by valvular cardiomyopathy, heart failure caused by other types of heart disease, heart failure with preserved systolic function (HFpEF), heart failure with intermediate systolic function (HFmEF), or heart failure with reduced systolic function (HFrEF).

[0018] In some embodiments, the heart failure patient suffers from heart failure caused by dilated cardiomyopathy.

[0019] In some embodiments, the heart failure is not right heart failure.

[0020] In some embodiments, the heart failure is left heart failure. In some embodiments, the heart failure is global heart failure.

[0021] In some embodiments, the heart failure is acute heart failure. In some embodiments, the heart failure is chronic heart failure. In some embodiments, the heart failure is acute-on-chronic heart failure or acute-onset heart failure.

[0022] In some embodiments, the heart failure is heart failure caused by ischemic myocardial damage, heart failure caused by hypertension, heart failure caused by alcoholic cardiomyopathy, heart failure caused by dilated cardiomyopathy, heart failure caused by hypertrophic cardiomyopathy, heart failure caused by valvular cardiomyopathy, or heart failure caused by other types of heart disease.

[0023] In some embodiments, the heart failure is heart failure with preserved systolic function (HFpEF), heart failure with intermediate systolic function (HFmEF), or heart failure with reduced systolic function (HFrEF).

[0024] In some embodiments, a PKM2 level in a sample from a subject that is elevated by more than 10% relative to a predetermined standard level indicates that the subject may be suffering from heart failure. In some embodiments, a PKM2 level in a sample from a subject that is elevated by more than 20% relative to a predetermined standard level indicates that the subject may be suffering from heart failure. In some embodiments, a PKM2 level in a sample from a subject that is elevated by more than 30% relative to a predetermined standard level indicates that the subject may be suffering from heart failure. In some embodiments, a PKM2 level in a sample from a subject that is elevated by more than 50% relative to a predetermined standard level indicates that the subject may be suffering from heart failure. In some embodiments, a PKM2 level in a sample from a subject that is elevated by more than 60% relative to a predetermined standard level indicates that the subject may be suffering from heart failure. In some embodiments, a PKM2 level in a sample from a subject that is elevated by more than 70% relative to a predetermined standard level indicates that the subject may be suffering from heart failure. In some embodiments, a PKM2 level in a sample from a subject that is elevated by more than 80% relative to a predetermined standard level indicates that the subject may be suffering from heart failure. In some embodiments, a PKM2 level in a sample from a subject that is elevated by more than 90% relative to a predetermined standard level indicates that the subject may be suffering from heart failure. In some embodiments, a PKM2 level in a sample from a subject that is elevated by more than 100% relative to a predetermined standard level indicates that the subject may be suffering from heart failure. In some embodiments, a PKM2 level in a sample from a subject that is elevated by more than 150% relative to a predetermined standard level indicates that the subject may be suffering from heart failure. In some embodiments, a PKM2 level in a sample from a subject that is elevated by more than 200% relative to a predetermined standard level indicates that the subject may be suffering from heart failure.

[0025] In some embodiments, the PKM2 level in a subject sample or patient sample is determined using a PKM2 detection kit comprising a reagent capable of recognizing PKM2. In some embodiments, the reagent capable of recognizing PKM2 comprises an antibody or antigen-binding fragment thereof that specifically binds to PKM2 or a biologically active fragment thereof.

[0026] In some embodiments, the predetermined standard level of PKM2 is a PKM2 level from a population of healthy individuals without heart failure.

[0027] In some embodiments, the predetermined standard level of PKM2 is less than 10 U / ml. In some embodiments, the predetermined standard level of PKM2 is selected from 7 to 10 U / ml. In some embodiments, the predetermined standard level of PKM2 is selected from 7.5 U / ml, 7.6 U / ml, 7.7 U / ml, 7.8 U / ml, 7.9 U / ml, 8.0 U / ml, 8.1 U / ml, 8.2 U / ml, 8.3 U / ml, 8.4 U / ml, 8.5 U / ml, 8.6 U / ml, 8.7 U / ml, 8.8 U / ml, 8.9 U / ml, 9.0 U / ml, 9.1 U / ml, 9.2 U / ml, 9.3 U / ml, 9.4 U / ml, 9.5 U / ml, 9.6 U / ml, 9.7 U / ml, 9.8 U / ml, 9.9 U / ml. When the test results show that the subject's PKM2 level is higher than 10U / ml, 11U / ml, 12U / ml, 13U / ml, 14U / ml, 15U / ml, 16U / ml, 17U / ml, 18U / ml, 19U / ml, 20U / ml, 21U / ml, 22U / ml, 23U / ml, 24U / ml, 25U / ml, 26U / ml, 27U / ml, 28U / ml, 29U / ml, 30U / ml, 31U / ml, 32U / ml, 33U / ml, 34U / ml, 35U / ml, 36U / ml, 37U / ml, 38U / ml, 39U / ml or 40U / ml, it indicates that the subject may be a heart failure patient. When the test results show that the subject's PKM2 level is higher than 10U / ml, 11U / ml, 12U / ml, 13U / ml, 14U / ml, 15U / ml, 16U / ml, 17U / ml, 18U / ml, 19U / ml, 20U / ml, 21U / ml, 22U / ml, 23U / ml, 24U / ml, 25U / ml, 26U / ml, 27U / ml, 28U / ml, 29U / ml, 30U / ml, 31U / ml, 32U / ml, 33U / ml, 34U / ml or 35U / ml, it indicates that the subject may be a left heart failure patient. When the test results show that the subject's PKM2 level is higher than 10U / ml, 11U / ml, 12U / ml, 13U / ml, 14U / ml, 15U / ml, 16U / ml, 17U / ml, 18U / ml, 19U / ml, 20U / ml, 21U / ml, 22U / ml, 23U / ml, 24U / ml, 25U / ml, 26U / ml, 27U / ml, 28U / ml, 29U / ml, 30U / ml, 31U / ml, 32U / ml, 33U / ml, 34U / ml, 35U / ml, 36U / ml, 37U / ml, 38U / ml, 39U / ml or 40U / ml, it indicates that the subject may be a patient with total heart failure.

[0028] In some embodiments, the PKM2 level in the sample from an acute heart failure patient described in the present disclosure is between 10 and 100 U / ml. In some embodiments, the PKM2 level in the sample from an acute heart failure patient described in the present disclosure is between 10 and 80 U / ml. In some embodiments, the PKM2 level in the sample from an acute heart failure patient described in the present disclosure is between 10 and 60 U / ml. In some embodiments, the PKM2 level in the sample from an acute heart failure patient described in the present disclosure is between 10 and 40 U / ml. In some embodiments, the PKM2 level in the acute heart failure patient sample described in the present disclosure is higher than 10U / ml, 11U / ml, 12U / ml, 13U / ml, 14U / ml, 15U / ml, 16U / ml, 17U / ml, 18U / ml, 19U / ml, 20U / ml, 21U / ml, 22U / ml, 23U / ml, 24U / ml, 25U / ml, 26U / ml, 27U / ml, 28U / ml, 29U / ml, 30U / ml, 31U / ml, 32U / ml, 33U / ml, 34U / ml, 35U / ml, 36U / ml, 37U / ml, 38U / ml, 39U / ml or 40U / ml. In some embodiments, the PKM2 level in the chronic heart failure patient sample described in the present disclosure is between 10 and 30U / ml. In some embodiments, the PKM2 level in the chronic heart failure patient sample described herein is between 10 and 20 U / ml. In some embodiments, the PKM2 level in the chronic heart failure patient sample described herein is greater than 10 U / ml, 11 U / ml, 12 U / ml, 13 U / ml, 14 U / ml, 15 U / ml, 16 U / ml, 17 U / ml, 18 U / ml, 19 U / ml, or 20 U / ml.

[0029] The subject sample or patient sample of the present disclosure is a biological sample. In some embodiments, the subject sample or patient sample is a blood sample. In some embodiments, the subject sample or patient sample is a myocardial tissue sample. In some embodiments, the subject sample or patient sample is a serum sample. In some embodiments, the subject sample or patient sample is a plasma sample.

[0030] In some embodiments, the PKM2 level described herein is obtained by measuring the PKM2 content in plasma, wherein the plasma sample is obtained by collecting anticoagulated whole blood from a subject and centrifuging the whole blood to obtain plasma. In some embodiments, blood is collected from a subject into a blue-capped vacutainer (containing sodium citrate as an anticoagulant, with a ratio of anticoagulant to blood of 1:9) and the blood in the blood tube is centrifuged to obtain plasma.

[0031] In some embodiments, the subject or heart failure patient described herein is not a patient with a malignant tumor.

[0032] In some embodiments, the detection method includes but is not limited to colloidal gold immunochromatography, immunoblotting, immunohistochemistry, immunofluorescence, enzyme-linked immunosorbent assay, nucleic acid probe method or real-time quantitative q-PCR method.

[0033] As used herein, a "detectable increase" refers to an increase of 5% or more relative to a predetermined standard level of PKM2, for example, an increase of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 250%, or 300% or more.

[0034] Advantageous Effects of the Invention

[0035] The present disclosure found that the content of PKM2 in the plasma of heart failure patients is higher than that of non-heart failure patients, especially in patients with left heart failure and total heart failure, the content is significantly higher than that of non-heart failure patients, and the content in the plasma of patients with acute heart failure is significantly higher than that of non-heart failure patients. As a heart failure marker, PKM2 has high accuracy in diagnosing heart failure and prognosis of heart failure, providing a new and better option for the prediction, diagnosis, auxiliary diagnosis and prognosis of heart failure. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 : PKM2 expression levels in heart failure (HF) patients and normal non-HF patients;

[0037] Figure 2 : PKM2 expression levels in heart failure (HF) patients with different cardiac systolic function;

[0038] Figure 3 : PKM2 expression levels in patients with different types of heart failure (HF);

[0039] Figure 4 : PKM2 expression levels in patients with heart failure (HF) of different etiologies;

[0040] Figure 5 : Correlation between cardiac function and PKM2 expression in patients with heart failure (HF);

[0041] Figure 6 :Correlation between cardiac end-systolic diameter and PKM2 expression level in patients with heart failure (HF);

[0042] Figure 7 : Correlation between cardiac end-diastolic diameter and PKM2 expression level in patients with heart failure (HF);

[0043] Figure 8 :PKM2 expression in patients with acute and chronic heart failure;

[0044] Figure 9 :Comparison of the diagnostic ability of PKM2 with other heart failure blood markers;

[0045] Figure 10 : The ability of PKM2 to assess the prognosis of patients with heart failure. DETAILED DESCRIPTION

[0046] The PKM2 detection kit used in the present disclosure was purchased from Nanjing Novozyme Medical Technology Co., Ltd., pyruvate kinase isoenzyme 2 detection kit (enzyme-linked immunosorbent assay): EL101.

[0047] The embodiments of the present disclosure will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are only used to illustrate the present disclosure and should not be regarded as limiting the scope of the present disclosure.

[0048] Example 1: Analysis of PKM2 expression in the blood of patients with heart failure and those without heart failure

[0049] Patients with a clear history of malignant tumors were excluded. A total of 126 patients with new heart failure or worsening heart failure admitted to the hospital for drug treatment and 46 other cardiology patients without heart failure (coronary heart disease, hypertension, syncope, atrial fibrillation, premature ventricular contractions, and pulmonary hypertension) were consecutively collected. Fasting blood was collected on the morning of the next day of admission and centrifuged at 2,500g for 10 minutes to obtain plasma. The expression level of plasma PKM2 in the two groups of patients was detected by ELISA. The difference in PKM2 levels between the two groups was compared by T test. Results: Compared with patients without heart failure (NC), the expression level of PKM2 in patients with heart failure (HF) was significantly increased, as shown in the attached figure. Figure 1 shown.

[0050] All heart failure patients were classified according to heart failure type, and the differences in PKM2 levels among different types and different causes of heart failure were compared using one-way analysis of variance. Results: There was no significant difference in plasma PKM2 levels between patients with preserved systolic function (HFpEF), intermediate systolic function (HFmEF), and reduced systolic function (HFrEF) (see Appendix). Figure 2 ), while the plasma PKM2 level in patients with isolated right heart failure had no significant difference with that in the normal control group (see Appendix Figure 3 ), there was no significant difference in PKM2 levels in patients with heart failure of different etiologies (see Appendix Figure 4 ).

[0051] Pearson correlation analysis was used to analyze the correlation between PKM2 levels and cardiac structure and function in 126 patients with heart failure. Results: Plasma PKM2 levels were correlated with cardiac function in patients with heart failure (r = -0.335, p < 0.01), but not with end-systolic diameter (r = 0.01, p > 0.05) or end-diastolic diameter (r = 0.03, p > 0.05). (See Appendix Figure 5 、 6 、7).

[0052] Patients with a clear history of malignant tumors were excluded. A total of 54 patients with acute heart failure were consecutively collected. 17 patients with acute heart failure who were converted to stable chronic heart failure after treatment were enrolled. Fasting blood was collected and centrifuged at 2,500g for 10 minutes to obtain plasma. The expression level of plasma PKM2 in the two groups of patients was detected by ELISA. The difference in PKM2 levels between the two groups of heart failure patients was compared by T test. Results: Compared with patients with chronic heart failure (CHF), the expression level of PKM2 in patients with acute heart failure (AHF) was significantly higher, as shown in the attached figure. Figure 8 shown.

[0053] Example 2

[0054] Sixty-five patients admitted to the emergency department with acute dyspnea were consecutively enrolled (excluding any history of malignancy). Of these, 43 had acute heart failure and 22 had non-cardiac dyspnea (16 with respiratory diseases, including pneumothorax, acute asthma attack, emphysema, lung infection, pleural effusion, one with pulmonary embolism, two with pericardial effusion, and three with unexplained dyspnea). Anticoagulated whole blood was collected and centrifuged at 2,500 g for 10 minutes to obtain plasma. Plasma levels of PKM2, NT-proBNP, and ST-2 were measured by ELISA. The area under the receiver operating characteristic (ROC) curve was calculated to compare the diagnostic value of these three markers for the differential diagnosis of acute heart failure. Results: Compared with NT-proBNP (area under the ROC curve: 0.827, 95% CI (0.700-0.955)) and ST-2 (area under the ROC curve: 0.686, 95% CI (0.529-0.844)), PKM2 (area under the ROC curve: 0.910, 95% CI (0.818-1.00)) was significantly increased (p < 0.01) (see Appendix Figure 9 ).

[0055] Example 3

[0056] Excluding patients with a history of malignancy, 59 consecutive patients with dilated cardiomyopathy who were hospitalized for new-onset heart failure or worsening heart failure were enrolled. Fasting blood was collected the morning after admission and centrifuged at 2,500 g for 10 minutes to obtain plasma. Plasma PKM2 expression levels were measured using an ELISA. Heart failure patients were divided into two groups based on the median plasma PKM2 level (18.18 U / L) at admission: high PKM2 (>50% of the median) and low PKM2 (<50% of the median). A composite endpoint of cardiovascular death and heart failure rehospitalization within one year was used. All heart failure patients were followed for one year, and Cox survival analysis was used to compare the one-year event-free survival between the high-PKM2 and low-PKM2 groups. Results: The composite endpoint event rate was significantly higher in the high-PKM2 group compared with the low-PKM2 group (HR 4.82 (95% CI 1.18-14.18), p = 0.02). (See Appendix.) Figure 10 (Percent survival (%) refers to event-free survival rate).

Claims

1. Use of a reagent for detecting PKM2 or mRNA encoding PKM2 protein in the preparation of a kit for predicting, diagnosing, assisting in the diagnosis, or prognostic evaluation of heart failure, wherein the reagent is used to detect the level of PKM2 or mRNA encoding PKM2 protein in a sample from a subject and compare the level with a predetermined standard level; a detectable increase in the level of PKM2 or mRNA encoding PKM2 protein in the sample from the subject relative to the predetermined standard level indicates the possibility of heart failure. wherein the subject sample is a serum sample or a plasma sample, wherein the subject is not a patient with a malignant tumor, The heart failure is left heart failure or total heart failure.

2. Use of a reagent for detecting PKM2 or mRNA encoding PKM2 protein in the preparation of a kit for diagnosing or assisting in the diagnosis of heart failure, wherein the diagnosis or assisting in the diagnosis includes: a. measuring the level of PKM2 or mRNA encoding PKM2 protein in a sample obtained from a subject presenting symptoms of heart failure; and b. compared to a predetermined standard level of PKM2 or mRNA encoding PKM2 protein, wherein a detectable increase in the level of PKM2 or mRNA encoding PKM2 protein in the sample relative to the predetermined standard level indicates a likelihood of heart failure, wherein the subject sample is a serum sample or a plasma sample, wherein the subject is not a patient with a malignant tumor, The heart failure is left heart failure or total heart failure.

3. The use according to claim 2, wherein the heart failure symptoms are selected from dyspnea, cough, sputum, hemoptysis, fatigue, dizziness, palpitations, decreased exercise, oliguria, symptoms of renal damage, cold limbs, forced sitting, confusion, cyanosis of the lips, pale complexion, sweating, and irritability.

4. The method according to claim 3, wherein the dyspnea is acute dyspnea.

5. Use of a reagent for detecting PKM2 or mRNA encoding PKM2 protein in the preparation of a kit for prognostic assessment of heart failure patients who are receiving anti-heart failure treatment, wherein the prognostic assessment comprises: a. Measure the level of PKM2 or mRNA encoding PKM2 protein in patient samples collected upon admission; and b. comparing the level of PKM2 or mRNA encoding PKM2 protein in the patient at admission to the hospital with a reference value, wherein the patient sample is a serum sample or a plasma sample, wherein the patient is not a patient with a malignant tumor.

6. The use according to claim 5, wherein if the patient's PKM2 level is higher than 50% of the reference value, the patient's prognosis is poor; if the patient's PKM2 level is lower than 50% of the reference value, the patient's prognosis is good.

7. The use according to claim 5, wherein the heart failure patient is selected from heart failure caused by ischemic myocardial damage, heart failure caused by hypertension, heart failure caused by alcoholic cardiomyopathy, heart failure caused by dilated cardiomyopathy, heart failure caused by hypertrophic cardiomyopathy, heart failure caused by valvular cardiomyopathy, or heart failure caused by other types of heart disease.

8. The use according to any one of claims 1, 2, and 5, wherein the heart failure is selected from left heart failure, total heart failure, acute heart failure, chronic heart failure, heart failure caused by ischemic myocardial damage, heart failure caused by hypertension, heart failure caused by alcoholic cardiomyopathy, heart failure caused by dilated cardiomyopathy, heart failure caused by hypertrophic cardiomyopathy, heart failure caused by valvular cardiomyopathy, heart failure caused by other types of heart disease, heart failure with preserved systolic function, heart failure with intermediate systolic function, or heart failure with decreased systolic function.

9. The use according to any one of claims 1, 2, and 5, wherein the heart failure is heart failure with preserved systolic function, heart failure with intermediate systolic function, or heart failure with decreased systolic function.

10. The use according to any one of claims 1, 2, and 5, wherein the reagent for detecting PKM2 comprises an antibody or an antigen-binding fragment thereof that can specifically bind to PKM2 or a biologically active fragment thereof.

11. The use according to any one of claims 1, 2, and 5, wherein the reagent for detecting mRNA encoding PKM2 protein comprises specific primers for amplifying mRNA encoding PKM2 protein, or a specific probe or chip for detecting mRNA encoding PKM2 protein.

Images