Use of il-1α inhibitor in preparing medicament for treating leukemia therapy-induced cardiac injury

Abstract

Provided are use of an IL-1α inhibitor in preparing a medicament for treating leukemia therapy-induced cardiac injury and use of an IL1R1 antagonist in preparing a medicament for treating leukemia therapy-induced cardiac injury. The present disclosure can effectively reduce cardiovascular complications and the occurrence of long-term cardiovascular adverse events in the period of leukemia treatments, thereby helping alleviate the cardiac metabolic dysfunction and cardiac dysfunction after treatment.

Description

Application of IL-1α inhibitors in the preparation of drugs for treating leukemia-induced cardiac damage

[0001] This application claims the benefit of Chinese patent application No. 2024101907113, filed on February 20, 2024. The entire text of the aforementioned Chinese patent application is incorporated herein by reference. Technical Field

[0002] The present invention relates to the field of biomedicine, and in particular to the use of an IL-1α inhibitor in the preparation of a medicine for treating heart damage induced by leukemia treatment. Background Art

[0003] Tumor diseases and cardiovascular diseases are two major types of diseases that seriously threaten human health. Leukemia accounts for one-third of childhood malignant tumor patients and is the most common childhood malignant tumor. Data show that with the emergence of new treatment methods and the optimization of treatment plans, the five-year overall survival rate of childhood acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML) has reached 90% and 78.2% respectively, which has continuously expanded the base of leukemia survivors. A recent clinical study shows that the long-term disease burden of leukemia survivors in childhood tumor diseases is much higher than that of survivors of other types of tumors, and the main burden is cardiovascular disease. 1 The incidence of heart disease in children with AML is significantly higher than that in children with ALL, mainly manifested as cardiomyopathy and heart failure. 2 It is generally believed that the risk of death from heart disease in leukemia survivors is mainly due to tumor chemotherapy. For example, in the 1970s, a study reported that anthracycline chemotherapy drugs, the core drugs in childhood leukemia chemotherapy regimens, have clear dose-dependent cardiac toxicity. 3 A study on childhood AML survivors showed that 12% of survivors will develop cardiac dysfunction within 5 years, of which about 70% will develop cardiac dysfunction during the peri-chemotherapy period. It is particularly important that whether treatment-related cardiac toxicity occurs in the early stage determines the long-term survival rate. 4 Therefore, early identification of chemotherapy-induced cardiotoxicity and active intervention may reduce the burden of cardiovascular disease. 5 .

[0004] Energy metabolism is the most important physiological activity of the heart. Many studies have found that abnormal energy metabolism is an important cause of heart damage. Neubauer et al. first proposed the concept that the heart with heart failure is a "fuel-depleted machine" 6However, it is not clear whether chemotherapy drugs can damage the heart by interfering with myocardial energy metabolism. On the other hand, previous studies in the field of oncology cardiology mainly used healthy mice to observe the direct damage to the heart caused by drugs after long-term, high-dose, multi-course chemotherapy. For example, Wallace et al. found that anthracyclines can damage myocardial cells by promoting the production of reactive oxygen species in the mitochondrial respiratory chain. 6 Zhang et al. found that anthracyclines can cause cardiac damage by inhibiting the activity of topoisomerase IIβ. 7 However, clinically, the inventors have found that children with leukemia can develop chemotherapy-related heart damage during the short-term induction chemotherapy phase. In addition, previous studies have ignored the indirect toxic effects of changes in the tumor microenvironment after chemotherapy on the heart, and have overlooked the possible interactions between tumors, chemotherapy drugs, and the heart. These issues prompted the inventors to consider whether there are unidentified mechanisms of chemotherapy-related heart damage, and then to reduce the burden of cardiovascular disease in children with leukemia after chemotherapy through precise intervention. Summary of the Invention

[0005] In order to address the deficiency in the prior art of lacking a method for accurately and effectively improving cardiac damage caused by leukemia treatment, such as cardiac metabolic disorders and abnormal cardiac function, the present invention provides the use of IL-1α inhibitors in the preparation of drugs for treating leukemia treatment-induced cardiac damage.

[0006] To solve the above technical problems, one of the technical solutions provided by the present invention is: use of an IL-1α (i.e., interleukin 1α) inhibitor in the preparation of a drug for treating leukemia-induced cardiac damage.

[0007] In a specific embodiment of the present invention, the leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy.

[0008] In a specific embodiment of the present invention, the immunotherapy is CAR-T therapy.

[0009] In a specific embodiment of the present invention, the immunotherapy is CD19 CAR-T therapy.

[0010] In a specific embodiment of the invention, the leukemia treatment results in necrosis of the leukemia cells.

[0011] In a specific embodiment of the present invention, the chemotherapy drug is an anthracycline chemotherapy drug.

[0012] In a specific embodiment of the present invention, the anthracycline chemotherapy drug is daunorubicin.

[0013] In a specific embodiment of the present invention, the IL-1α inhibitor is an anti-IL-1α antibody.

[0014] In a specific embodiment of the present invention, the anti-IL-1α antibody is a monoclonal antibody.

[0015] In a specific embodiment of the present invention, the anti-IL-1α antibody is a neutralizing antibody.

[0016] In a specific embodiment of the present invention, the anti-IL-1α antibody was purchased from R&D Systems with the product number AB-400-NA.

[0017] In a specific embodiment of the present invention, the leukemia is acute myeloid leukemia or acute lymphoblastic leukemia.

[0018] In a specific embodiment of the present invention, the acute lymphoblastic leukemia is acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia.

[0019] In a specific embodiment of the present invention, the acute myeloid leukemia is caused by MLL-AF9 fusion protein.

[0020] In a specific embodiment of the present invention, the cardiac injury is selected from one or more of: cardiometabolic disorders, abnormal cardiac function, cardiomyopathy and heart failure.

[0021] In a specific embodiment of the present invention, the cardiometabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

[0022] To solve the above technical problems, the second technical solution provided by the present invention is: use of an antibody purchased from R&D Systems with the product number AB-400-NA in the preparation of a drug for treating leukemia-induced cardiac damage.

[0023] In a specific embodiment of the present invention, the leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy; and / or, the leukemia is acute myeloid leukemia or acute lymphoblastic leukemia.

[0024] In a specific embodiment of the present invention, the immunotherapy is CAR-T therapy.

[0025] In a specific embodiment of the present invention, the immunotherapy is CD19 CAR-T therapy.

[0026] In a specific embodiment of the present invention, the acute lymphoblastic leukemia is acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia.

[0027] In a specific embodiment of the invention, the leukemia treatment results in necrosis of the leukemia cells.

[0028] In a specific embodiment of the present invention, the chemotherapy drug is an anthracycline chemotherapy drug, preferably daunorubicin; and / or the acute myeloid leukemia is caused by MLL-AF9 fusion protein.

[0029] In a specific embodiment of the present invention, the cardiac injury is selected from one or more of: cardiometabolic disorders, abnormal cardiac function, cardiomyopathy and heart failure.

[0030] In a specific embodiment of the present invention, the cardiometabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

[0031] To solve the above technical problems, the third technical solution provided by the present invention is: the use of an antibody purchased from R&D Systems with the product number AB-400-NA in the preparation of a drug for treating leukemia-induced cardiac damage by inhibiting IL-1α.

[0032] In a specific embodiment of the present invention, the leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy; and / or, the leukemia is acute myeloid leukemia or acute lymphoblastic leukemia.

[0033] In a specific embodiment of the present invention, the immunotherapy is CAR-T therapy.

[0034] In a specific embodiment of the present invention, the immunotherapy is CD19 CAR-T therapy.

[0035] In a specific embodiment of the present invention, the acute lymphoblastic leukemia is acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia.

[0036] In a specific embodiment of the invention, the leukemia treatment results in necrosis of the leukemia cells.

[0037] In a specific embodiment of the present invention, the chemotherapy drug is an anthracycline chemotherapy drug, preferably daunorubicin; and / or the acute myeloid leukemia is caused by MLL-AF9 fusion protein.

[0038] In a specific embodiment of the present invention, the cardiac injury is selected from one or more of: cardiometabolic disorders, abnormal cardiac function, cardiomyopathy and heart failure.

[0039] In a specific embodiment of the present invention, the cardiometabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

[0040] To solve the above technical problems, the fourth technical solution provided by the present invention is: use of an antagonist of IL1R1 (i.e., interleukin-1 receptor type I) in the preparation of a drug for treating leukemia-induced cardiac damage.

[0041] The present invention has demonstrated that knockout of the cardiac IL1R1 receptor can protect cardiac function (Example 2). Based on this, those skilled in the art can reasonably anticipate that IL1R1 antagonists can treat leukemia-induced cardiac damage.

[0042] In a specific embodiment of the present invention, the antagonist of IL1R1 is an anti-IL1R1 drug.

[0043] In a specific embodiment of the present invention, the antagonist of IL1R1 is a competitive inhibitory drug or a blocking drug of IL1R1.

[0044] In a specific embodiment of the present invention, the competitive inhibitory drug is Anakinra (Anakinra, a recombinant, non-glycosylated form of human interleukin-1 receptor antagonist (IL-1Ra), CAS No.: 143090-92-0).

[0045] In a specific embodiment of the present invention, the leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy.

[0046] In a specific embodiment of the present invention, the immunotherapy is CAR-T therapy.

[0047] In a specific embodiment of the present invention, the immunotherapy is CD19 CAR-T therapy. In a specific embodiment of the present invention, the chemotherapy drug is an anthracycline chemotherapy drug, such as daunorubicin; and / or the leukemia treatment causes leukemia cell necrosis.

[0048] In a specific embodiment of the present invention, the leukemia is acute myeloid leukemia or acute lymphocytic leukemia; and / or the cardiac injury is selected from one or more of: cardiac metabolic disorders, abnormal cardiac function, cardiomyopathy and heart failure.

[0049] In a specific embodiment of the present invention, the acute lymphoblastic leukemia is acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia.

[0050] In a specific embodiment of the present invention, the acute myeloid leukemia is caused by MLL-AF9 fusion protein; and / or the cardiac metabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

[0051] To solve the above technical problems, the fifth technical solution provided by the present invention is: a method for treating leukemia-induced cardiac damage, the method comprising administering a therapeutically effective amount of an IL-1α inhibitor to a patient in need.

[0052] In a specific embodiment of the present invention, the leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy.

[0053] In a specific embodiment of the present invention, the immunotherapy is CAR-T therapy.

[0054] In a specific embodiment of the present invention, the immunotherapy is CD19 CAR-T therapy. In a specific embodiment of the present invention, the leukemia treatment causes necrosis of leukemia cells.

[0055] In a specific embodiment of the present invention, the chemotherapy drug is an anthracycline chemotherapy drug.

[0056] In a specific embodiment of the present invention, the anthracycline chemotherapy drug is daunorubicin.

[0057] In a specific embodiment of the present invention, the IL-1α inhibitor is an anti-IL-1α antibody.

[0058] In a specific embodiment of the present invention, the anti-IL-1α antibody is a monoclonal antibody.

[0059] In a specific embodiment of the present invention, the anti-IL-1α antibody is a neutralizing antibody.

[0060] In a specific embodiment of the present invention, the anti-IL-1α antibody was purchased from R&D Systems with the product number AB-400-NA.

[0061] In a specific embodiment of the present invention, the leukemia is acute myeloid leukemia or acute lymphoblastic leukemia.

[0062] In a specific embodiment of the present invention, the acute lymphoblastic leukemia is acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia.

[0063] In a specific embodiment of the present invention, the acute myeloid leukemia is caused by MLL-AF9 fusion protein.

[0064] In a specific embodiment of the present invention, the cardiac injury is selected from one or more of: cardiometabolic disorders, abnormal cardiac function, cardiomyopathy and heart failure.

[0065] In a specific embodiment of the present invention, the cardiometabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

[0066] In a specific embodiment of the invention, the patient is a child.

[0067] To solve the above technical problems, the sixth technical solution provided by the present invention is: a method for treating cardiac damage induced by leukemia treatment, the method comprising administering a therapeutically effective amount of an antibody purchased from R&D Systems with the product number AB-400-NA to a patient in need.

[0068] In a specific embodiment of the present invention, the leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy; and / or, the leukemia is acute myeloid leukemia or acute lymphoblastic leukemia.

[0069] In a specific embodiment of the present invention, the immunotherapy is CAR-T therapy.

[0070] In a specific embodiment of the present invention, the immunotherapy is CD19 CAR-T therapy.

[0071] In a specific embodiment of the present invention, the acute lymphoblastic leukemia is acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia.

[0072] In a specific embodiment of the invention, the leukemia treatment results in necrosis of the leukemia cells.

[0073] In a specific embodiment of the present invention, the chemotherapy drug is an anthracycline chemotherapy drug, preferably daunorubicin; and / or the acute myeloid leukemia is caused by MLL-AF9 fusion protein.

[0074] In a specific embodiment of the present invention, the cardiac injury is selected from one or more of: cardiometabolic disorders, abnormal cardiac function, cardiomyopathy and heart failure.

[0075] In a specific embodiment of the present invention, the cardiometabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

[0076] In a specific embodiment of the invention, the patient is a child.

[0077] To solve the above technical problems, the seventh technical solution provided by the present invention is: a method for treating leukemia-induced cardiac damage, comprising administering a therapeutically effective amount of an IL1R1 antagonist to a patient in need.

[0078] In a specific embodiment of the present invention, the antagonist of IL1R1 is an anti-IL1R1 drug.

[0079] In a specific embodiment of the present invention, the antagonist of IL1R1 is a competitive inhibitory drug or a blocking drug of IL1R1.

[0080] In a specific embodiment of the present invention, the competitive inhibitory drug is Anakinra.

[0081] In a specific embodiment of the present invention, the leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy.

[0082] In a specific embodiment of the present invention, the immunotherapy is CAR-T therapy.

[0083] In a specific embodiment of the present invention, the immunotherapy is CD19 CAR-T therapy. In a specific embodiment of the present invention, the chemotherapy drug is an anthracycline chemotherapy drug, such as daunorubicin; and / or the leukemia treatment causes leukemia cell necrosis.

[0084] In a specific embodiment of the present invention, the leukemia is acute myeloid leukemia or acute lymphocytic leukemia; and / or the cardiac injury is selected from one or more of: cardiac metabolic disorders, abnormal cardiac function, cardiomyopathy and heart failure.

[0085] In a specific embodiment of the present invention, the acute lymphoblastic leukemia is acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia.

[0086] In a specific embodiment of the present invention, the acute myeloid leukemia is caused by MLL-AF9 fusion protein; and / or the cardiac metabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

[0087] In a specific embodiment of the invention, the patient is a child.

[0088] As used herein, the term "effective amount" refers to an amount of a drug or pharmaceutical agent that elicits the biological or pharmaceutical response of a tissue, system, animal, or human that is being sought, for example, by a researcher or clinician. Additionally, the term "therapeutically effective amount" refers to an amount that results in improved treatment, cure, prevention, or alleviation of a disease, condition, or side effect, or that reduces the rate of progression of a disease or condition, compared to a corresponding subject that has not received that amount. The term also includes within its scope amounts that are effective to enhance normal physiological function.

[0089] To solve the above technical problems, the eighth technical solution provided by the present invention is: an IL-1α inhibitor for treating heart damage induced by leukemia treatment.

[0090] In a specific embodiment of the present invention, the leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy.

[0091] In a specific embodiment of the present invention, the immunotherapy is CAR-T therapy.

[0092] In a specific embodiment of the present invention, the immunotherapy is CD19 CAR-T therapy.

[0093] In a specific embodiment of the invention, the leukemia treatment results in necrosis of the leukemia cells.

[0094] In a specific embodiment of the present invention, the chemotherapy drug is an anthracycline chemotherapy drug.

[0095] In a specific embodiment of the present invention, the anthracycline chemotherapy drug is daunorubicin.

[0096] In a specific embodiment of the present invention, the IL-1α inhibitor is an anti-IL-1α antibody.

[0097] In a specific embodiment of the present invention, the anti-IL-1α antibody is a monoclonal antibody.

[0098] In a specific embodiment of the present invention, the anti-IL-1α antibody is a neutralizing antibody.

[0099] In a specific embodiment of the present invention, the anti-IL-1α antibody was purchased from R&D Systems with the product number AB-400-NA.

[0100] In a specific embodiment of the present invention, the leukemia is acute myeloid leukemia or acute lymphoblastic leukemia.

[0101] In a specific embodiment of the present invention, the acute lymphoblastic leukemia is acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia.

[0102] In a specific embodiment of the present invention, the acute myeloid leukemia is caused by MLL-AF9 fusion protein.

[0103] In a specific embodiment of the present invention, the cardiac injury is selected from one or more of: cardiometabolic disorders, abnormal cardiac function, cardiomyopathy and heart failure.

[0104] In a specific embodiment of the present invention, the cardiometabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

[0105] To solve the above technical problems, the ninth technical solution provided by the present invention is: an antibody purchased from R&D Systems with the product number AB-400-NA, which is used to treat heart damage induced by leukemia treatment.

[0106] In a specific embodiment of the present invention, the leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy; and / or, the leukemia is acute myeloid leukemia or acute lymphoblastic leukemia.

[0107] In a specific embodiment of the present invention, the immunotherapy is CAR-T therapy.

[0108] In a specific embodiment of the present invention, the immunotherapy is CD19 CAR-T therapy.

[0109] In a specific embodiment of the present invention, the acute lymphoblastic leukemia is acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia.

[0110] In a specific embodiment of the invention, the leukemia treatment results in necrosis of the leukemia cells.

[0111] In a specific embodiment of the present invention, the chemotherapy drug is an anthracycline chemotherapy drug, preferably daunorubicin; and / or the acute myeloid leukemia is caused by MLL-AF9 fusion protein.

[0112] In a specific embodiment of the present invention, the cardiac injury is selected from one or more of: cardiometabolic disorders, abnormal cardiac function, cardiomyopathy and heart failure.

[0113] In a specific embodiment of the present invention, the cardiometabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

[0114] To solve the above technical problems, the tenth technical solution provided by the present invention is: an antagonist of IL1R1, which is used to treat heart damage induced by leukemia treatment.

[0115] In a specific embodiment of the present invention, the antagonist of IL1R1 is an anti-IL1R1 drug.

[0116] In a specific embodiment of the present invention, the antagonist of IL1R1 is a competitive inhibitory drug or a blocking drug of IL1R1.

[0117] In a specific embodiment of the present invention, the competitive inhibitory drug is Anakinra.

[0118] In a specific embodiment of the present invention, the leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy.

[0119] In a specific embodiment of the present invention, the immunotherapy is CAR-T therapy.

[0120] In a specific embodiment of the present invention, the immunotherapy is CD19 CAR-T therapy.

[0121] In a specific embodiment of the present invention, the chemotherapy drug is an anthracycline chemotherapy drug, such as daunorubicin; and / or the leukemia treatment causes leukemia cell necrosis.

[0122] In a specific embodiment of the present invention, the leukemia is acute myeloid leukemia or acute lymphocytic leukemia; and / or the cardiac injury is selected from one or more of: cardiac metabolic disorders, abnormal cardiac function, cardiomyopathy and heart failure.

[0123] In a specific embodiment of the present invention, the acute lymphoblastic leukemia is acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia.

[0124] In a specific embodiment of the present invention, the acute myeloid leukemia is caused by MLL-AF9 fusion protein; and / or the cardiac metabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

[0125] On the basis of conforming to the common sense in this field, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0126] The reagents and raw materials used in the present invention are commercially available.

[0127] The positive progress effect of the present invention is:

[0128] The present invention is the first to propose that the release of tumor-derived cytokine IL-1α caused by leukemia treatment (such as chemotherapy or immunotherapy) is the key to causing cardiac energy metabolism remodeling and functional abnormalities, emphasizing that the interaction between drugs and tumors plays an important role in the cardiotoxicity of leukemia treatment drugs (such as chemotherapy drugs or CAR-T). The use of the IL-1α inhibitor provided by the present invention in the preparation of a drug for treating leukemia treatment-induced cardiac damage will effectively reduce cardiovascular complications during leukemia treatment, reduce the incidence of long-term adverse cardiovascular events, and help improve cardiac metabolic disorders and cardiac function abnormalities after treatment. It is likely to significantly improve the long-term prognosis of leukemia patients, benefiting many leukemia patients, thereby achieving significant social and economic effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0129] Figure 1 shows the construction of cardiac-specific Il1r1 knockout mice. (Top) Schematic diagram of the construction; (Bottom) Western blot analysis to verify the IL1R1 knockout effect. The control group was IL1R1 fl / fl, and the experimental group was IL1R1 cKO. The protein expression levels of IL1R1 relative to Actin are shown on the right. IL1R1: interleukin-1 receptor type 1; Actin: actin. Statistical differences between the two groups were calculated using a two-independent t-test ( * P<0.05).

[0130] Figure 2 shows the construction of AML cells with stable knockdown of Il1a. (Top) Construction diagram; (Bottom) RT-PCR verification of knockout effect. The control group used the scramble sequence, and the experimental group used the shIL-1α sequence. The right side shows the mRNA expression of Il1a relative to Gapdh. The statistical difference between the two groups of samples was calculated using a two-independent sample t-test ( *** P<0.001).

[0131] Figure 3 demonstrates the effect of myocardial-specific overexpression of PGC-1α. The control group was AAV9-NC, and the experimental group was AAV9-PGC1α. The right side shows the protein expression of PGC-1α relative to Actin. NC: negative control, PGC-1α: peroxisome proliferator-activated receptor γ coactivator 1-α, Actin: actin. The statistical difference between the two groups was calculated using a two-independent sample t-test ( ** P<0.01).

[0132] FIG4 shows the p65 overexpression plasmid and the PGC-1α promoter fluorescence reporter plasmid.

[0133] Figure 5 is a schematic diagram of the construction of the MLL-AF9 AML mouse model. AML: acute myeloid leukemia.

[0134] Figure 6A-Figure 6D show the changes in cardiac metabolism and function in AML mice after DNR chemotherapy.

[0135] Figure 6A shows that AML mice were divided into two groups, one group received DNR chemotherapy (AML+DNR group), and the other group received PBS as a control (AML group).

[0136] FIG6B shows cardiac ultrasound examination of the two groups of mice.

[0137] Figure 6C shows the cardiac PET-CT examination of the two groups of mice. The signal strengths of the fluorine-18 labeled FTHA and FDG probes reflect the levels of fatty acid and glucose utilization by the heart, respectively.

[0138] Figure 6D shows the evaluation of myocardial ATP content in the two groups of mice.

[0139] AML: acute myeloid leukemia; DNR: daunorubicin; LVEF: left ventricular ejection fraction; LVFS: left ventricular short-axis contraction fraction; FDG: deoxyglucose; FTHA: 6-thio-heptadecanoic acid; ATP: adenosine triphosphate; The statistical differences between the two groups were calculated using the two-independent sample t test ( * P < 0.05; ** P<0.01 *** P<0.001).

[0140] Figure 7 shows decreased expression of key enzymes of cardiac lipid metabolism in leukemic mice after chemotherapy. AML: acute myeloid leukemia; DNR: daunorubicin; CD36: leukocyte differentiation antigen 36; CPT1: carnitine palmitoyltransferase 1; LCAD: long-chain acyl-CoA dehydrogenase; MCAD: medium-chain acyl-CoA dehydrogenase; Actin: actin. Statistical differences between the two groups were calculated using a two-sample t-test (*P < 0.05; **P < 0.01; ***P < 0.001).

[0141] Figures 8A-8D show single-cell sequencing analysis of the myocardium of AML children after DNR chemotherapy and normal myocardium.

[0142] FIG8A is a UMAP image showing the cluster distribution of cardiac cells.

[0143] Figure 8B shows the scMetabolism metabolic scores of normal myocardium and myocardial cells after chemotherapy; the carbohydrate metabolism pathway is highlighted in red, and the fatty acid metabolism pathway is highlighted in blue.

[0144] FIG8C shows a pseudo-time series analysis of all cardiomyocytes.

[0145] FIG8D shows a pseudo-sequential branch point analysis of all cardiomyocytes.

[0146] The names of genes related to fatty acid metabolism that are downregulated in non-adapted cardiomyocytes are highlighted in blue, and the names of genes related to glucose metabolism that are upregulated are highlighted in red.

[0147] Figure 9A-Figure 9D show the changes in cardiac metabolism and function in normal tumor-free mice after DNR chemotherapy.

[0148] Figure 9A shows that healthy tumor-free mice were divided into two groups, one group received DNR chemotherapy (TF+DNR group), and the other group received PBS (TF group) as a control.

[0149] FIG9B shows cardiac ultrasound examination of the two groups of mice.

[0150] FIG9C shows the cardiac PET-CT examination of the two groups of mice.

[0151] FIG9D shows the detection of myocardial ATP content in two groups of mice.

[0152] TF: tumor-free; DNR: daunorubicin; LVEF: left ventricular ejection fraction; LVFS: left ventricular short-axis contraction fraction; FTHA: 6-thio-heptadecanoic acid; FDG: deoxyglucose; ATP: adenosine triphosphate; the statistical differences between the two groups were calculated using the two-sample t-test (NS: not significant).

[0153] FIG10A-FIG10C are cardiac transcriptomic analyses of AML mice after DNR chemotherapy.

[0154] Figure 10A shows the KEGG pathway enrichment analysis of differentially expressed genes in cardiac transcriptomics results after chemotherapy. The cytokine and cytokine interaction pathways were significantly enriched and highlighted in red.

[0155] Figure 10B shows the biological response pathway analysis of cardiac transcriptomics results after chemotherapy, with PPAR signaling significantly downregulated and highlighted in blue.

[0156] Figure 10C is a protein interaction network analysis of differentially expressed genes in the PPAR pathway, showing that IL-1 signaling and NF-KappaB signaling closely interact with the key metabolic protein PPARGC1A.

[0157] Figures 11A-11D show the detection of multiple factors in plasma of AML mice after DNR chemotherapy, and the boxes indicate IL-1α.

[0158] TF: tumor-free; AML: acute myeloid leukemia; DNR: daunorubicin. Differences between samples were calculated using analysis of variance followed by the Tukey-Kramer test (NS: not significant; *P < 0.05; **P < 0.01; ***P < 0.001).

[0159] Figures 12A-12C show that AML cells undergo necrosis and release IL-1α after DNR chemotherapy.

[0160] FIG12A and FIG12B are flow cytometric measurements of the ratio of live cells to necrotic cells in primary AML cell lines after 24 h of intervention with different DNR concentration gradients.

[0161] FIG12C shows the detection of IL-1α content in the cell culture supernatant after 24 h of 0.5 μM DNR intervention in the primary AML cell line.

[0162] IL-1α: interleukin 1α; DNR: daunorubicin; the statistical differences between two groups of samples were calculated using the two-independent sample t-test, and the differences between more than two groups of samples were calculated using the post-hoc Tukey-Kramer test for analysis of variance (*P<0.05; **P<0.01; ***P<0.001).

[0163] 13A-13D show the changes in cardiac metabolism and function in normal tumor-free mice after administration of IL-1α.

[0164] FIG13A shows healthy tumor-free mice divided into two groups, one group receiving IL-1α (TF+IL-1α group) and the other group receiving PBS as a control (TF group).

[0165] FIG13B shows cardiac ultrasound examination of the two groups of mice.

[0166] FIG13C shows the cardiac PET-CT examination of the two groups of mice.

[0167] FIG13D shows the detection of myocardial ATP content in two groups of mice.

[0168] TF: tumor-free; LVEF: left ventricular ejection fraction; LVFS: left ventricular short-axis contraction fraction; FTHA: 6-thio-heptadecanoic acid; FDG: deoxyglucose; ATP: adenosine triphosphate; The statistical differences between the two groups were calculated using the two-independent sample t test ( * P < 0.05; ** P<0.01 *** P<0.001).

[0169] Figure 14A-Figure 14C show that cardiac-specific Il1r1 knockout improves cardiac metabolism and function in mice after chemotherapy.

[0170] FIG14A is a schematic diagram of the experiment, showing DNR intervention in AML mice with cardiomyocyte-specific knockout of IL1R1 (IL1R1cKO group) and control AML mice (IL1R1 fl / fl group).

[0171] FIG14B shows cardiac ultrasound examination of the two groups of mice.

[0172] FIG14C shows the detection of myocardial ATP content in two groups of mice.

[0173] AML: acute myeloid leukemia; DNR: daunorubicin; LVEF: left ventricular ejection fraction; LVFS: left ventricular fractional contraction; ATP: adenosine triphosphate; The statistical differences between the two groups were calculated using the two-independent-sample t-test (***P<0.001).

[0174] Figure 15 shows the correlation analysis between the increase rate of plasma IL-1α and the decrease rate of LVEF in patients with acute myeloid leukemia before and after chemotherapy. LVEF: left ventricular ejection fraction.

[0175] Figure 16A-Figure 16C show that knockdown of Il1a in AML cells improves cardiac metabolism and function in mice after chemotherapy.

[0176] Figure 16A is a schematic diagram of the experiment, in which DNR intervention was performed on AML models constructed using control (scramble group) and IL-1α knockdown (shIL-1α group) cells.

[0177] FIG16B shows cardiac ultrasound examination of the two groups of mice.

[0178] FIG16C shows the detection of myocardial ATP content in two groups of mice.

[0179] WT: wild type; AML: acute myeloid leukemia; DNR: daunorubicin; LVEF: left ventricular ejection fraction; LVFS: left ventricular short-axis contraction fraction; ATP: adenosine triphosphate; the statistical differences between the two groups were calculated using the two-independent sample t test ( *** P<0.001).

[0180] Figure 17 shows the detection of the cardiac NF-KappaB signaling pathway in normal tumor-free mice and AML mice after DNR chemotherapy. (Top) Immunoblotting detection of the cardiac NF-KappaB pathway in normal tumor-free mice after chemotherapy; (Bottom) Immunoblotting detection of the cardiac NF-KappaB pathway in AML mice after chemotherapy. TF: tumor-free; AML: acute myeloid leukemia; DNR: daunorubicin; p-p65: phosphorylated transcription factor p65; p65: transcription factor p65; IκBα: nuclear factor κB inhibitor protein; Actin: actin. The statistical difference between the two groups of samples was calculated using a two-independent sample t-test (NS: no significant difference; * P < 0.05; ** P<0.01).

[0181] Figure 18 shows immunofluorescence and immunoblotting of myocardial tissues in AML patients treated with DNR chemotherapy and normal myocardium. (Top) Immunofluorescence analysis of IL-1α in myocardium after chemotherapy (patient) and normal myocardium (control); gray indicates ACTIN2, green indicates IL-1α, and blue indicates DAPI; scale bar: 50 μm; (Bottom) Immunoblotting analysis of the NF-κB signaling pathway in myocardium after chemotherapy and normal myocardium. PGC-1α: peroxisome proliferator-activated receptor γ coactivator 1-α; p65: transcription factor p65; p-p65: phosphorylated transcription factor p65; Actin: actin.

[0182] Figure 19 shows that an NF-κB signaling antagonist improves cardiac metabolism and function in mice after chemotherapy. (Top) Experimental schematic: DNR intervention in AML mice treated with PDTC (PDTC group) or untreated (control group); (Middle) Cardiac ultrasound examination of the two groups of mice; (Bottom) Myocardial ATP content in the two groups of mice. AML: acute myeloid leukemia; DNR: daunorubicin; PDTC: ammonium pyrrolidine dithiocarbamate; LVEF: left ventricular ejection fraction; LVFS: left ventricular short-axis contraction fraction; ATP: adenosine triphosphate; Statistical differences between the two groups were calculated using a two-sample t-test (**P < 0.01; ***P < 0.001).

[0183] Figure 20 shows the changes in the expression of PGC-1α in the heart of AML mice after DNR chemotherapy. (Top) Detection of PGC-1α protein expression in the heart of AML mice after chemotherapy. The right side shows the protein expression of PGC-1α relative to Actin; (Bottom) Detection of PGC-1α transcription level in the heart of AML mice after chemotherapy. AML: acute myeloid leukemia; DNR: daunorubicin; PGC-1α: peroxisome proliferator-activated receptor γ coactivator 1-α; Actin: actin. The statistical difference between the two groups of samples was calculated using a two-independent sample t-test ( ** P<0.01).

[0184] 21A-21D show the expression of myocardial PGC-1α in children with AML after DNR chemotherapy.

[0185] FIG21A is a UMAP image showing that PGC-1α is mainly expressed in cardiomyocytes in cardiac tissue.

[0186] FIG21B is a pseudo-time series analysis showing the cell fate of different cardiomyocyte subpopulations.

[0187] FIG21C shows the expression levels of PGC-1α in different cardiomyocyte subsets.

[0188] FIG21D is a pseudo-time series analysis showing the relationship between different cardiomyocyte fates and PGC-1α expression levels.

[0189] Figure 22 shows a dual luciferase reporter assay. Overexpression of p65 significantly inhibited the transcription of PGC-1α. TSS: transcription start site; LUC: firefly enzyme; RLU: relative fluorescence intensity; PGC-1α: peroxisome proliferator-activated receptor γ coactivator 1-α; p65: transcription factor p65 ( *** P<0.001).

[0190] Figure 23 shows that cardiac PGC-1α overexpression improves cardiac metabolism and function in mice after chemotherapy. (Top) Experimental diagram, DNR intervention in AML mice with cardiac overexpression of PGC-1α (AAV9-PGC1α group) and control AML mice (AAV9-NC group); (Middle) Cardiac ultrasound detection of the two groups of mice; (Bottom) Detection of myocardial ATP content in the two groups of mice. WT: wild type; AAV9: adeno-associated virus type 9; NC: negative control; PGC-1α: peroxisome proliferator-activated receptor γ coactivator 1-α; AML: acute myeloid leukemia; DNR: daunorubicin; LVEF: left ventricular ejection fraction; LVFS: left ventricular short-axis contraction rate; ATP: adenosine triphosphate; The statistical differences between the two groups of samples were calculated using a two-independent sample t-test ( * P < 0.05; *** P<0.001).

[0191] FIG24A and FIG24B show that IL-1α neutralizing antibodies improve cardiac metabolism and function in mice after chemotherapy.

[0192] Figure 24A (top) Schematic diagram of the experiment, DNR intervention in AML mice treated with IL-1α neutralizing antibody (IL-1αAb group) or isotype control antibody (IgG group); (middle) cardiac ultrasound detection of the two groups of mice; (bottom) detection of myocardial ATP content in the two groups of mice.

[0193] Figure 24B shows the percentage of residual GFP+ tumor cells in the bone marrow of the two groups of mice on day 4 after chemotherapy.

[0194] AML: acute myeloid leukemia; DNR: daunorubicin; Ab: antibody; LVEF: left ventricular ejection fraction; LVFS: left ventricular fraction; ATP: adenosine triphosphate; BM: bone marrow; GFP: green fluorescent protein; The statistical differences between the two groups were calculated using the two-sample t-test (NS: not significant; * P < 0.05; ** P<0.01).

[0195] Figure 25 illustrates the mechanism by which chemotherapy leads to cardiac metabolic disorders and functional impairment. Following chemotherapy, tumor cell necrosis releases IL-1α, which acts on the cardiac IL1R1 receptor, activating the NF-KappaB signaling pathway and inhibiting PGC-1α expression, leading to cardiac metabolic disorders and functional abnormalities.

[0196] Figure 26 shows that IL-1α neutralizing antibodies improve cardiac metabolism and function in T-ALL mice after chemotherapy. (Left) Experimental schematic: DNR intervention in T-ALL mice treated with IL-1α neutralizing antibodies (IL-1αAb group) or isotype control antibodies (IgG group); (Middle) Cardiac ultrasound examination of the two groups of mice; (Right) Myocardial ATP content in the two groups of mice. T-ALL: acute lymphoblastic T-cell leukemia; DNR: daunorubicin; LVEF: left ventricular ejection fraction; LVFS: left ventricular short-axis contraction fraction; ATP: adenosine triphosphate; Statistical differences between the two groups were calculated using a two-sample t-test (**P < 0.01; ***P < 0.001).

[0197] Figure 27 shows that IL-1α neutralizing antibodies improve cardiac metabolism and function in B-ALL mice after CAR-T therapy. (Left) Schematic diagram of the experiment: CD19 CAR-T cells were administered to B-ALL mice treated with either IL-1α neutralizing antibodies (IL-1αAb group) or isotype control antibodies (IgG group); (Middle) Cardiac ultrasound examination of the two groups of mice; (Right) Myocardial ATP content was measured in the two groups of mice. B-ALL: acute lymphoblastic B-cell leukemia; CAR-T: chimeric antigen receptor T cell immunotherapy; LVEF: left ventricular ejection fraction; LVFS: left ventricular short-axis contraction fraction; ATP: adenosine triphosphate; Statistical differences between the two groups were calculated using a two-sample t-test (*P < 0.05; **P < 0.01).

[0198] Figure 28 shows that Anakinra improves cardiac metabolism and function in mice after chemotherapy. (Left) Schematic diagram of the experiment: DNR intervention in AML mice treated with Anakinra (Anakinra group) or not treated with Anakinra (control group); (Middle) Cardiac ultrasound examination of the two groups of mice; (Right) Myocardial ATP content in the two groups of mice. AML: acute myeloid leukemia; DNR: daunorubicin; LVEF: left ventricular ejection fraction; LVFS: left ventricular fractional contraction; ATP: adenosine triphosphate; Statistical differences between the two groups were calculated using a two-sample t-test (***P < 0.001). DETAILED DESCRIPTION

[0199] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0200] Experimental methods

[0201] Mouse cardiac function assessment

[0202] The Fujifilm Vevo3100 small animal ultrasound system enables high-resolution imaging of the heart. This study used ultrasound to assess cardiac function in leukemia mice before and after chemotherapy, clarifying the changes in cardiac function caused by leukemia chemotherapy. More precise cardiac function data was acquired using a 9.4T Bruker BioSpin small animal magnetic resonance imaging system, equipped with a mouse-specific cardiac coil and cardiac gating device, enabling accurate assessment of mouse cardiac function.

[0203] Cardiac isotope tracer metabolic flux detection

[0204] Cardiac isotope tracer metabolic flux detection can accurately reflect the specific flow direction and flux of glycolipid metabolic substrates in cardiac energy metabolism. The present invention uses C13-labeled glucose and palmitic acid from Sigma. According to the method reported by Motoaki Sano et al. (J Mol Cell Cardiol. 2015 May; 82: 116-24.), a C13-glucose injection dose of 1 mg / g was selected. According to the BSA heating dissolution method reported by Kim et al. (Sci Rep. 2017; 7: 4335.), and based on the tolerance of mice, a C13-palmitic acid injection dose of 0.5 mg / g was selected. The mouse hearts were removed 15 minutes after injection for subsequent mass spectrometry analysis.

[0205] Preparatory Example

[0206] Construction of a leukemia mouse therapeutic model

[0207] AML mouse model construction:

[0208] The leukemia mouse chemotherapy model used in this invention is based on the transfection of primary myeloid stem / progenitor cells from normal mice with a lentiviral-packaged MLL-AF9 plasmid. GFP+ leukemia cells are then screened by flow cytometry and further transplanted into irradiated mice in which myeloid cells have been eliminated for expansion, thereby creating MLL-AF9 AML mice. This model preserves the immune function of the recipient mice and can simulate the pathophysiological process of childhood leukemia. It is currently a recognized AML model in the field of hematological tumors. Mice that receive leukemia cell bone marrow transplants develop leukemia symptoms such as splenomegaly and fatigue approximately 20-30 days after transplantation.

[0209] Construction of T-ALL mouse model (reference: PD-1 signaling defines and protects leukaemic stem cells from T cell receptor-induced cell death in T cell acute lymphoblastic leukaemia. DOI: 10.1038 / s41556-022-01050-3):

[0210] The T-ALL mouse model used is based on transfection of primary myeloid stem / progenitor cells from normal mice with a lentiviral NOTCH1 plasmid. GFP+ leukemic cells are then selected by flow cytometry and expanded by transplantation into irradiated mice. Overactivating mutations in NOTCH1 occur in over 60% of human T-ALL.

[0211] Construction of B-ALL mouse model (reference: Ink4a and Arf are crucial factors in the determination of the cell of origin and the therapeutic sensitivity of Myc-induced mouse lymphoid tumor.DOI:10.1038 / onc.2011.462):

[0212] The B-ALL mouse model used is based on transfection of primary hematopoietic stem cells of normal mice with lentiviral-packaged N-Myc plasmid, followed by flow cytometry screening of GFP+ leukemia cells, which are further transplanted into irradiated mice for expansion, thereby constructing B-ALL mice.

[0213] The chemotherapy regimen used in the present invention is based on the anthracycline dosage for clinical leukemia children during the induction chemotherapy phase, and adopts the method of administering 5 mg / kg of DNR for two consecutive days for chemotherapy.

[0214] The CAR-T therapy regimen used in the present invention (reference: Therapeutic efficacy of anti-CD19 CAR-T cells in a mouse model of systemic lupus erythematosus. DOI: 10.1038 / s41423-020-0472-1):

[0215] 1. Chimeric Antigen Receptor Composition: The variable regions of the light and heavy chains of the 1D3 anti-CD19 antibody were used as scFv fragments, with mCD8 as the transmembrane region, and m4-1BB and mCD3ζ signaling as the cytoplasmic region. These sequences were synthesized and ligated into a lentiviral vector carrying an IRES-EGFP fluorescent signal.

[0216] 2. After lentivirus infection of T cells, CD19 CAR-T cells were obtained.

[0217] 3. 1E6 CAR-T cells were administered to each mouse for two consecutive days (B-ALL mice were cleared of lymphocytes).

[0218] Construction of cardiac-specific Il1r1 knockout mice and systemic Il1a knockout mice

[0219] Il1r1-Flox mice were purchased from Shanghai Model Organisms Technology Co., Ltd. The flox site was inserted into the 3-4 exon region of the Il1r1 gene in this strain of mice. Il1r1 was then constructed by mating with Myh6-CreERT2 transgenic mice. flox / flox Myh6-CreERT2 mice were used. Immunoblotting revealed a significant decrease in IL1R1 protein expression in myocardial tissue of mice treated with 100 mg / kg tamoxifen for 5 days and a 3-week follow-up, confirming the effectiveness of this genetically engineered mouse strain (Figure 1). Systemic Il1a knockout mice were purchased from Shanghai Model Organisms Co., Ltd. This strain contains exons 5-6 of the Il1a gene, and their genotype has been verified by sequencing.

[0220] Il1a knockdown AML cell construction

[0221] Using molecular cloning techniques, the commercially available Sigma MISSION shRNA plasmid was modified to include a red fluorescent protein (RFP) gene sequence and a shRNA targeting mouse Il1a. The plasmid was then packaged into lentiviral vectors and transfected into primary AML cells. Finally, GFP+RFP+ cells were flow-sorted. RT-PCR confirmed that Il1a expression was significantly reduced in these GFP+RFP+ AML cells, demonstrating the successful establishment of an AML cell line with stable Il1a knockdown (Figure 2).

[0222] AAV9-PGC-1α virus construction

[0223] AAV9-PGC-1α virus carrying a myocardial specific promoter was purchased from Shanghai Hanheng Biotechnology Co., Ltd. In the present invention, the applicant used the tail vein injection method, and the number of viruses was selected as 4*10 13vg / kg dose injection (Gene Ther. 2011 Jan; 18(1): 43-52). Three weeks after the virus injection, immunoblotting confirmed that the expression of PGC-1α in the heart of wild-type mice was significantly increased (Figure 3).

[0224] Construction of p65 overexpression plasmid and PGC-1α promoter fluorescence reporter plasmid

[0225] The present invention cloned the amplified full-length human p65 coding sequence (NM_021975) into the CV702 vector, and simultaneously cloned the human PGC-1α (NM_013261) promoter sequence covering a 1100 bp fragment (from -1000 bp to +100 bp) into the GV238 luciferase vector ( FIG4 ).

[0226] Example 1: Leukemia chemotherapy induces cardiac energy metabolism disorder and heart damage

[0227] The heart is an organ with a high energy demand. It beats more than 100,000 times a day, transports 10 tons of blood to the body, and consumes 6 kilograms of ATP. 6 In the fasting state, 70% of the heart's energy is supplied by fatty acids, so energy conversion is the most core metabolic process of the heart. Heart energy metabolism in heart failure will be reshaped, from fatty acid energy supply to glucose energy supply, accompanied by decreased cardiac ATP synthesis and dysfunction. 8 Numerous studies have confirmed the close connection between cardiac energy metabolism disorders and heart damage. For example, in a recent study, the applicant found that energy imbalance induced by insulin resistance in adolescence directly leads to heart damage and dysfunction. 9 However, is the cardiac toxicity experienced by children with leukemia after chemotherapy related to abnormal energy metabolism? Can the cardiovascular damage caused by chemotherapy be alleviated by improving cardiac energy metabolism? These questions are still unclear and require further exploration.

[0228] It is generally believed that gene fusion is the main pathogenesis of childhood leukemia 10 MLL-AF9 fusion protein is present in 25% of newly diagnosed AML children 11, the AML mouse model carrying this fusion protein is widely used in the field of pediatric hematological tumor research. The inventors transfected the MLL-AF9 fusion gene into mouse myeloid cells through retrovirus and constructed an acute myeloid leukemia (MLL-AF9 AML) mouse tumor-bearing model (Figure 5). Then, after the model mice showed leukemia-related signs, the inventors divided them into two groups. One group used the anthracycline chemotherapy drug daunorubicin (DNR) for intervention according to the clinical AML short-term induction chemotherapy regimen (5 mg / kg of DNR was given for two consecutive days), and the other group used phosphate buffered saline (PBS) as a control (Figure 6A). The inventors further used small animal cardiac ultrasound to detect changes in left ventricular ejection fraction (LVEF) and left ventricular short-axis contraction fraction (LVFS) in mice after chemotherapy. The results showed that compared with the control group, the cardiac function of leukemia mice showed a significant decline after short-term induction chemotherapy (Figure 6B). Then, the inventors used PET-CT to evaluate changes in cardiac energy metabolism in leukemia mice after chemotherapy. The results showed that compared with the control group, the heart of leukemia mice had reduced fatty acid uptake and enhanced glucose uptake after chemotherapy (Figure 6C). Furthermore, the inventors detected the ATP content in the myocardial tissue of leukemia mice after chemotherapy, and the results showed that the ATP content in the heart decreased significantly after chemotherapy (Figure 6D). Finally, the inventors used immunoblotting to evaluate the expression of lipid metabolism-related proteins, and the results showed that the expression of key enzymes of cardiac lipid metabolism was significantly reduced after chemotherapy (Figure 7). These results demonstrate that leukemia mice had fatty acid metabolism disorders after chemotherapy, accompanied by a decline in cardiac function. In order to further verify the inventors' conclusions in human specimens, the inventors collected myocardial biopsy specimens from children with AML who developed abnormal cardiac function after using DNR chemotherapy and normal myocardial specimens from children donated due to brain death, and performed single-cell transcriptome analysis. The results also showed that myocardial fatty acid metabolism in children decreased and glucose metabolism increased after chemotherapy (Figure 8A-Figure 8D). In summary, the inventors believe that cardiac energy metabolism remodeling is closely related to cardiac damage and decreased cardiac function caused by chemotherapy, and impaired fatty acid metabolism is an important cause of its cardiac dysfunction.

[0229] Example 2 Tumor cell-derived IL-1α induces cardiac metabolic and functional disorders

[0230] What factors drive the changes in cardiac energy metabolism in leukemia mice after chemotherapy? Studies have shown that in vitro perfusion of anthracyclines in the heart reduces carnitine content and thus affects cardiac metabolism. 12 Drugs approved for the treatment of metabolic diseases such as diabetes, such as metformin (MET) and empagliflozin (EMPA), can prevent anthracycline-induced cardiotoxicity. 13 On the other hand, the release of inflammatory factors such as interferon gamma (IFN-γ) triggered by meningococcal disease has also been reported to cause cardiac damage.14 So is it the chemotherapy drugs themselves that cause cardiac energy metabolism disorders in mice? Or is it certain toxic cytokines released after chemotherapy destroys tumor cells that lead to abnormal cardiac energy metabolism?

[0231] In order to exclude the influencing factors of leukemia cells, the inventors first referred to the induction chemotherapy regimen of the previous experiment to perform chemotherapy in normal mice without leukemia, and observed the direct effects of chemotherapy drugs on the cardiac energy metabolism and cardiac function of mice. The inventors divided healthy tumor-free mice into two groups, one of which used DNR chemotherapy and the other used PBS as a control, and then performed cardiac ultrasound and PET-CT detection (Figure 9A). Interestingly, the inventors found that the cardiac metabolism and function of healthy tumor-free mice did not change significantly after chemotherapy (Figure 9B-Figure 9C), and the content of cardiac ATP did not show a significant decrease (Figure 9D). Therefore, the cardiac energy metabolism disorder after chemotherapy may not be a direct effect of chemotherapy drugs, but a comprehensive result of the interaction between chemotherapy drugs and leukemia cells. Further, the inventors performed transcriptomic sequencing on the heart tissue of leukemia mice after chemotherapy. The results showed that the cytokine and cytokine receptor interaction pathway was significantly enriched in the myocardial tissue after chemotherapy (Figure 10A-Figure 10C). This result suggests that chemotherapy may cause certain tumor-derived cytokines to be released into the blood, leading to energy metabolism disorders in the heart.

[0232] Based on the above conjecture, the inventors used Luminex multifactor detection technology to perform high-throughput screening of cytokines that changed in the plasma of leukemia mice and normal tumor-free mice after chemotherapy. The results showed that among all the cytokines detected, 9 cytokines were significantly increased in leukemia mice after chemotherapy. Since the specific cytokines released by tumor cells need to conform to the change pattern of increasing after chemotherapy in leukemia mice but not significantly increasing after chemotherapy in healthy tumor-free mice, the inventors verified these 9 cytokines one by one, and the results showed that only interleukin 1α (IL-1α) met this trend characteristic (Figure 11A-Figure 11D). The IL-1 family is divided into two subtypes: IL-1α and IL-1β. In the field of cardiovascular disease research, many studies have reported the effects of IL-1β on cardiomyocytes. 15 However, research on IL-1α has mainly focused on its effects on cardiac vascular endothelial cells and fibroblasts, and there are few studies on its direct effects on cardiomyocytes. It is currently believed that IL-1α is a damage-related pattern molecule that is constitutively expressed in blood or non-blood cells. After cell necrosis, IL-1α is released into the intercellular space or blood circulation to exert its biological functions locally or systemically. 16. The inventors' previous in vitro experiments showed that DNR can induce necrosis of leukemia cells, and the necrosis ratio increased in a dose-dependent manner (Figure 12A and Figure 12B). In addition, the inventors also found that the protein expression of IL-1α in the leukemia cell culture supernatant was significantly increased after the addition of DNR (Figure 12C). Therefore, IL-1α released by leukemia cell necrosis caused by chemotherapy drugs may be a potential cause of cardiac metabolic disorders and cardiac function damage.

[0233] In order to verify the role of IL-1α in cardiac metabolism and function, the inventors directly injected IL-1α recombinant protein into leukemia-free mice and found that IL-1α had a direct damaging effect on the heart (Figure 13A-Figure 13D). The inventors further used cardiac-specific IL-1α receptor (IL1R1) knockout mice to construct a leukemia model and found that cardiac IL-1α receptor knockout protected the cardiac metabolism and function of leukemia mice after chemotherapy. The above two experiments confirmed the key role of IL-1α in cardiac damage after chemotherapy from both positive and negative aspects (Figure 14A-Figure 14C). At the same time, the inventors collected blood samples from children with leukemia after chemotherapy, evaluated their plasma IL-1α concentration and cardiac function before and after chemotherapy, and proved that the increase in IL-1α in children was correlated with the decline in cardiac function (Figure 15).

[0234] To verify the source of IL-1α, the inventors used systemic IL-1α knockout mice to construct a leukemia model. At the same time, the inventors knocked down the IL-1α gene in primary leukemia cells using short hairpin RNA (shRNA) technology, and then constructed a leukemia model using the IL-1α knockdown cell line. Finally, by detecting the changes in plasma IL-1α levels in the two models after chemotherapy, it was confirmed that the increase in plasma IL-1α after chemotherapy was derived from the destruction and release of tumor cells (Figures 16A-16C).

[0235] Example 3 NF-KappaB signaling pathway mediates IL-1α-induced cardiac injury

[0236] So how does the increase in plasma IL-1α levels after chemotherapy cause cardiometabolic disorders? Existing studies have shown that activation of the IL-1α receptor IL1R1 can further activate the downstream NF-KappaB signaling pathway, thereby regulating gene expression. 17 It is generally believed that NF-KappaB signaling is a common pathway for many inflammatory cytokines to exert their biological functions, and plays an important role in the progression of myocardial infarction and heart failure. The NF-KappaB protein family exists in the form of a heterodimeric complex, of which p50 / p65 is the most important NF-KappaB complex in the heart. The p65 subunit is a transcription factor that can enter the cell nucleus after phosphorylation to regulate the transcription of downstream genes. 18Recent studies have shown that p65 plays an important role in the regulation of multiple metabolic genes in cells.

[0237] The transcriptomic results suggest that the NF-KappaB signaling pathway in the heart of leukemia mice is significantly enriched after chemotherapy (Figure 10A). Furthermore, the inventors preliminarily verified the activation level of the NF-KappaB signaling pathway in the heart of leukemia mice after chemotherapy by immunoblotting. The results showed that the level of p65 phosphorylation in the heart increased, and the expression level of the NF-KappaB inhibitory protein IkBα decreased. However, there was no significant change in the activation level of the NF-KappaB signaling pathway in the heart of healthy tumor-free mice after chemotherapy (Figure 17). In addition, the inventors previously performed immunofluorescence and immunoblotting tests on myocardial tissue after chemotherapy in leukemia children. The results showed that IL-1α was enriched in the myocardial tissue gap after chemotherapy, accompanied by an increase in the activation level of the NF-KappaB signaling pathway (Figure 18). Therefore, the inventors speculated that the increase in plasma IL-1α caused by chemotherapy may have activated the NF-KappaB signaling pathway in myocardial cells, thereby affecting the expression of downstream metabolic genes.

[0238] To verify this hypothesis, the inventors used PDTC, an inhibitor of the NF-KappaB signaling pathway, to intervene in leukemia mice. Compared with the control group, the cardiac metabolism and function of the PDTC intervention group were significantly improved after chemotherapy ( FIG19 ).

[0239] Example 4 PGC-1α reduction is the core of IL-1α-mediated cardiometabolic disorders

[0240] PGC-1α is a transcriptional coactivator that can bind to multiple transcription factors and play an important role in fatty acid oxidation, glycolysis, mitochondrial biogenesis, etc. 19 In recent years, PGC-1α has been found to be closely related to cardiac energy metabolism. Studies have shown that decreased expression of PGC-1α can inhibit cardiac fatty acid metabolism and further induce cardiac dysfunction. 20 Studies have shown that activation of the NF-KappaB signaling pathway can downregulate the expression of PGC-1α in cells, ultimately changing the cell metabolic pattern. 21 .

[0241] In previous studies, the inventors used the Ingenuity Pathway Analysis system to enrich the cardiac transcriptomic data of leukemia mice after chemotherapy. The results showed that the peroxisome proliferator-activated receptor (PPAR) signaling pathway, which is closely related to metabolism, was significantly downregulated (Figure 10B). Furthermore, the inventors performed a protein interaction network analysis on the differential genes in the PPAR pathway and found that PGC-1α and p65 interacted with each other in the central genes (Figure 10C). Then, the inventors detected the expression changes of PGC-1α at the transcriptional and translational levels in the hearts of leukemia mice after chemotherapy by RT-PCR and immunoblotting. The results showed that the expression of PGC-1α in the hearts of leukemia mice was significantly downregulated after chemotherapy (Figure 20). In addition, the inventors' previous single-cell transcriptomics results showed that the expression level of PGC-1α in cardiomyocytes of leukemia children decreased after chemotherapy, and its expression level determined the different fates of cardiomyocytes after chemotherapy (Figure 21A-Figure 21D). Therefore, the inventors speculate that the activation of the myocardial NF-KappaB signaling pathway after chemotherapy may cause cardiac energy metabolism disorders and functional damage by inhibiting the expression of PGC-1α.

[0242] The inventors used a dual-luciferase reporter assay to demonstrate that p65 can bind to the promoter region of the PGC-1α gene, thereby inhibiting PGC-1α transcription (Figure 22). Using adeno-associated virus type 9 (AAV9-PGC-1α) carrying a myocardial-specific promoter and the PGC-1α gene coding sequence to specifically overexpress PGC-1α in the hearts of leukemia mice, they corrected the cardiac energy metabolism disorder and functional abnormalities after chemotherapy, further demonstrating the central role of PGC-1α in chemotherapy-related cardiac dysfunction (Figure 23).

[0243] Example 5 IL-1α neutralizing antibodies alleviate cardiac damage and metabolic disorders induced by leukemia treatment

[0244] After clarifying the mechanism by which leukemia treatment (chemotherapy or immunotherapy, etc.) causes cardiac damage, the inventors believe that IL-1α neutralizing antibodies have translational value and broad application prospects for improving cardiac damage in treated patients. IL-1α plays a key role in both leukemia development and cardiac damage, and blocking the IL-1α-mediated cardiac NF-kappaB / PGC-1α axis may reduce the potential risk of cardiotoxicity.

[0245] The present invention found that the use of a 10 μg / mouse dose of IL-1α neutralizing antibody (Mouse IL-1alpha / IL-1F1 antib ody, catalog number: AB-400-NA, supplier: R&D Systems, product website: https: / / www.rndsystems.com / cn / products / mouse-il-1alpha-il-1f1-antibody_ab-400-na#product-details) can reduce cardiac metabolic and functional damage in leukemia mice (AML, T-ALL or B-ALL) after treatment without affecting the therapeutic effect ( Figures 24A and 24B, Figures 26 and 27 ).

[0246] Example 6 IL1R1 antagonists alleviate cardiac damage and metabolic disorders induced by leukemia treatment

[0247] The present invention found that the use of an IL1R1 antagonist (Anakinra, CAS No.: 143090-92-0) at a dose of 30 mg / kg / d×5 days can alleviate cardiac metabolic and functional damage in leukemia mice (AML mice) after treatment without affecting the therapeutic effect ( Figure 28 ).

[0248] summary

[0249] The introduction of the concept of "oncological cardiology" has made the complications of cardiac damage caused by anti-tumor treatment more and more concerned. However, many studies currently focus on cardiac damage directly caused by chemotherapy drugs, but ignore the interaction between tumors and chemotherapy drugs, and the differences in tumor load levels that may cause cardiac damage after chemotherapy. The present invention found that there is a close relationship between tumor cells, chemotherapy drugs, and cardiac damage. Chemotherapy releases IL-1α by inducing tumor cell necrosis, which acts on the myocardial IL1R1 receptor and activates the NF-KappaB signaling pathway, thereby inhibiting the expression of PGC-1α and ultimately leading to cardiac energy metabolism disorders and abnormal cardiac function (Figure 25).

[0250] The number of childhood leukemia survivors is increasing year by year, and life expectancy is continuously increasing. The resulting long-term cardiovascular burden continues to increase, and cardiovascular health problems caused by chemotherapy seriously affect their quality of life. Currently, there is a lack of effective protective strategies for leukemia chemotherapy-related cardiac damage. Early identification of cardiac pathological and physiological changes caused by leukemia chemotherapy and active intervention will effectively reduce the risk of cardiovascular diseases in children with leukemia after chemotherapy. It is urgent to develop personalized peri-chemotherapy cardiovascular protection strategies based on the unique spectrum of tumor diseases and physiological characteristics of children. This will effectively reduce cardiovascular complications during chemotherapy and reduce the incidence of long-term adverse cardiovascular events, benefiting many children with leukemia and thus achieving significant social and economic effects.

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Claims

1. Application of IL-1α inhibitors in the preparation of drugs for treating leukemia-induced cardiac damage.

2. The use according to claim 1, characterized in that The leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy; Preferably, the leukemia treatment satisfies one or more of the following conditions: The chemotherapy drug is an anthracycline chemotherapy drug, such as daunorubicin; The leukemia treatment results in necrosis of leukemia cells; and, The immunotherapy is CAR-T therapy, such as CD19 CAR-T therapy.

3. The use according to claim 1, characterized in that The leukemia is acute myeloid leukemia or acute lymphocytic leukemia, such as acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia; and / or the cardiac injury is selected from one or more of: cardiac metabolic disorder, abnormal cardiac function, cardiomyopathy and heart failure; Preferably, the acute myeloid leukemia is caused by MLL-AF9 fusion protein; and / or the cardiac metabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

4. The use according to claim 1, wherein The IL-1α inhibitor is an anti-IL-1α antibody; Preferably, the anti-IL-1α antibody is a monoclonal antibody and / or a neutralizing antibody; More preferably, the anti-IL-1α antibody is purchased from R&D Systems with the product number AB-400-NA.

5. Use of the antibody AB-400-NA purchased from R&D Systems in the preparation of a drug for the treatment of leukemia-induced cardiac damage; Preferably, the leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy; and / or, the leukemia is acute myeloid leukemia or acute lymphoblastic leukemia, such as acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia; More preferably, the leukemia treatment causes leukemia cell necrosis; and / or, the immunotherapy is CAR-T therapy, such as CD19 CAR-T therapy.

6. The use according to claim 5, characterized in that The chemotherapy drug is an anthracycline chemotherapy drug, preferably daunorubicin; and / or the acute myeloid leukemia is caused by MLL-AF9 fusion protein.

7. The use according to claim 5, characterized in that The cardiac injury is selected from one or more of: cardiac metabolic disorder, abnormal cardiac function, cardiomyopathy and heart failure; Preferably, the cardiometabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

8. Use of IL1R1 antagonists in the preparation of drugs for treating leukemia-induced cardiac damage; Preferably, the IL1R1 antagonist is a competitive inhibitory drug or a blocking drug of IL1R1; More preferably, the competitive inhibitory drug is Anakinra.

9. The use according to claim 8, characterized in that The leukemia treatment method is selected from: one or more of chemotherapy, radiotherapy, molecular targeted therapy and immunotherapy; Preferably, the leukemia treatment satisfies one or more of the following conditions: The chemotherapy drug is an anthracycline chemotherapy drug, such as daunorubicin; The leukemia treatment results in necrosis of leukemia cells; and, The immunotherapy is CAR-T therapy, such as CD19 CAR-T therapy.

10. The use according to claim 8, characterized in that The leukemia is acute myeloid leukemia or acute lymphocytic leukemia, such as acute lymphoblastic T-cell leukemia or acute lymphoblastic B-cell leukemia; and / or the cardiac injury is selected from one or more of: cardiac metabolic disorder, abnormal cardiac function, cardiomyopathy and heart failure; Preferably, the acute myeloid leukemia is caused by MLL-AF9 fusion protein; and / or the cardiac metabolic disorder is decreased myocardial fatty acid metabolism and enhanced glucose metabolism.

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