TECHNICAL FIELD

The present technology relates generally to compositions and methods for detecting and diagnosing diseases and conditions associated with defects in cardiolipin remodeling and/or mitochondrial dysfunction. In particular, the present technology relates to methods for detecting the presence or amount of cardiolipin isoforms and/or the presence or amount of enzymes involved in cardiolipin remodeling.

BACKGROUND

The following description is provided to assist the understanding of the reader. None of the information provided or references cited is admitted to be prior art to the present invention.

Heart failure is a leading cause of mortality and morbidity worldwide. In the United States, it affects nearly 5 million people and is the only major cardiovascular disorder on the rise. It is estimated that 400,000 to 700,000 new cases of heart failure are diagnosed each year in the U.S. and the number of deaths in the U.S. attributable to this condition has more than doubled since 1979, currently averaging 250,000 annually. Although heart failure affects people of all ages, the risk of heart failure increases with age and is most common among older people. Accordingly, the number of people living with heart failure is expected to increase significantly as the elderly population grows over the next few decades. The causes of heart failure have been linked to various disorders including coronary artery disease, atherosclerosis, past myocardial infarction, hypertension, abnormal heart valves, cardiomyopathy or myocarditis, congenital heart disease, severe lung disease, diabetes, severe anemia, hyperthyroidism, arrhythmia or dysrhythmia.

Multiple forms of heart failure are associated with defects in remodeling of the mitochondrial phospholipid cardiolipin, including genetic and sporadic forms of heart failure. Defects in cardiolipin remodeling thus serve as a diagnostic marker for heart failure, which may be monitored via the presence or level of cardiolipin isoforms present in a subject, and/or the presence or level of enzymes involved in cardiolipin remodeling.

SUMMARY

In one aspect, the present disclosure provides a method for diagnosing heart failure in a mammalian subject, the method comprising assessing cardiolipin remodeling in a biological sample from the subject.

In some embodiments, assessing cardiolipin remodeling comprises detecting levels of cardiolipin remodeling enzymes.

In some embodiments, detecting levels of cardiolipin remodeling enzymes comprises detecting the level of one or more of TAZ1, MLCL AT1, or ALCAT1 mRNA compared to a normal control subject.

In some embodiments, the level of MLCL AT1 or ALCAT1 mRNA is elevated about 2.5-fold compared to a normal control subject.

In some embodiments, the level of TAZ1 mRNA is reduced about 2.5-fold compared to a normal control subject.

In some embodiments, assessing cardiolipin remodeling comprises detecting levels of cardiolipin isoforms compared to a normal control subject.

In some embodiments, detecting one or more of TAZ1, MLCL AT1, or ALCAT1 mRNA comprises RT-PCR, in situ hybridization, or Northern blotting.

In some embodiments, detecting levels of cardiolipin isoforms comprises chromatography, mass spectrometry, ELISA, Western blotting, immunodetection, or immunoprecipitation.

In some embodiments, assessing cardiolipin remodeling comprises a mitochondrial function assay.

In some embodiments, the mitochondrial function assay comprises the use of peripheral blood mononuclear cells (PBMCs), leukocytes, or ex vivo tissues.

In some embodiments, the method further comprises administering to the subject a therapeutically effective amount of the peptide D-Arg-2′6′-Dmt-Lys-Phe-NH2or a pharmaceutically acceptable salt thereof.

In some embodiments, the peptide is administered daily for 6 weeks or more.

In some embodiments, the peptide is administered daily for 12 weeks or more.

In some embodiments, the heart failure results from hypertension; ischemic heart disease; exposure to a cardiotoxic compound; myocarditis; thyroid disease; viral infection; gingivitis; drug abuse; alcohol abuse; pericarditis; atherosclerosis; vascular disease; hypertrophic cardiomyopathy; acute myocardial infarction; left ventricular systolic dysfunction; coronary bypass surgery; starvation; an eating disorder; or a genetic defect.

In some embodiments, the subject is human.

In some embodiments, the peptide is administered orally, topically, systemically, intravenously, subcutaneously, intraperitoneally, or intramuscularly

In some embodiments, the method further comprises separately, sequentially or simultaneously administering a cardiovascular agent to the subject.

In some embodiments, the cardiovascular agent is selected from the group consisting of: an anti-arrhythmia agent, a vasodilator, an anti-anginal agent, a corticosteroid, a cardioglycosidc, a diuretic, a sedative, an angiotcnsin converting enzyme (ACE) inhibitor, an angiotensin II antagonist, a thrombolytic agent, a calcium channel blocker, a throboxane receptor antagonist, a radical scavenger, an anti-platelet drug, a β-adrenaline receptor blocking drug, α-receptor blocking drug, a sympathetic nerve inhibitor, a digitalis formulation, an inotrope, and an antihyperlipidemic drug.

In some embodiments, the pharmaceutically acceptable salt comprises acetate or trifluoroacetate salt.

In some embodiments, the biological sample comprises tissue, a cell, or a mitochondria from the subject.

In one aspect, the present disclosure provides a method for monitoring treatment for heart failure in a mammalian subject in need thereof, the method comprising assessing cardiolipin remodeling in a biological sample from the subject.

In some embodiments, assessing cardiolipin remodeling comprises detecting levels of cardiolipin remodeling enzymes.

In some embodiments, detecting levels of cardiolipin remodeling enzymes comprises detecting the level of one or more of TAZ1, MLCL AT1, or ALCAT1 mRNA compared to a normal control subject.

In some embodiments, the level of MLCL AT1 or ALCAT1 mRNA is elevated about 2 to about 2.5-fold compared to a normal control subject.

In some embodiments, the level of TAZ1 mRNA is reduced about 2 to about 2.5-fold compared to a normal control subject.

In some embodiments, assessing cardiolipin remodeling comprises detecting levels of cardiolipin isoforms compared to a normal control subject.

In some embodiments, detecting one or more of TAZ1, MLCL AT1, or ALCAT1 mRNA comprises RT-PCR, in situ hybridization, or Northern blotting.

In some embodiments, detecting levels of cardiolipin isoforms comprises chromatography, mass spectrometry, ELISA, Western blotting, immunodetection, or immunoprecipitation.

In some embodiments, assessing cardiolipin remodeling comprises a mitochondrial function assay.

In some embodiments, the mitochondrial function assay comprises the use of peripheral blood mononuclear cells (PBMCs), leukocytes, or ex vivo tissues.

In some embodiments, the method further comprises administering to the subject a therapeutically effective amount of the peptide D-Arg-2′6′-Dmt-Lys-Phe-NH2or a pharmaceutically acceptable salt thereof.

In some embodiments, the peptide is administered daily for 6 weeks or more.

In some embodiments, the peptide is administered daily for 12 weeks or more.

In some embodiments, the heart failure results from hypertension; ischemic heart disease; exposure to a cardiotoxic compound; myocarditis; thyroid disease; viral infection; gingivitis; drug abuse; alcohol abuse; pericarditis; atherosclerosis; vascular disease; hypertrophic cardiomyopathy; acute myocardial infarction; left ventricular systolic dysfunction; coronary bypass surgery; starvation; an eating disorder; or a genetic defect.

In some embodiments, the subject is human.

In some embodiments, the peptide is administered orally, topically, systemically, intravenously, subcutaneously, intraperitoneally, or intramuscularly

In some embodiments, the method further comprises separately, sequentially or simultaneously administering a cardiovascular agent to the subject.

In some embodiments, the cardiovascular agent is selected from the group consisting of: an anti-arrhythmia agent, a vasodilator, an anti-anginal agent, a corticosteroid, a cardioglycoside, a diuretic, a sedative, an angiotensin converting enzyme (ACE) inhibitor, an angiotensin II antagonist, a thrombolytic agent, a calcium channel blocker, a throboxane receptor antagonist, a radical scavenger, an anti-platelet drug, a β-adrenaline receptor blocking drug, α-receptor blocking drug, a sympathetic nerve inhibitor, a digitalis formulation, an inotrope, and an antihyperlipidemic drug.

In some embodiments, the pharmaceutically acceptable salt comprises acetate or trifluoroacetate salt.

In some embodiments, the biological sample comprises tissue, a cell, or a mitochondria from the subject.

In one aspect, the present disclosure provides a method for assessing mitochondrial dysfunction in a mammalian subject in need thereof, the method comprising assessing cardiolipin remodeling in a biological sample from the subject.

In some embodiments, assessing cardiolipin remodeling comprises detecting levels of cardiolipin remodeling enzymes.

In some embodiments, detecting levels of cardiolipin remodeling enzymes comprises detecting the level of one or more of TAZ1, MLCL AT1, or ALCAT1 mRNA compared to a normal control subject.

In some embodiments, the level of MLCL AT1 or ALCAT1 mRNA is elevated about 2.5-fold compared to a normal control subject.

In some embodiments, the level of TAZ1 mRNA is reduced about 2 to about 2.5-fold compared to a normal control subject.

In some embodiments, assessing cardiolipin remodeling comprises detecting levels of cardiolipin isoforms compared to a normal control subject.

In some embodiments, detecting one or more of TAZ1, MLCL AT1, or ALCAT1 mRNA comprises RT-PCR, in situ hybridization, or Northern blotting.

In some embodiments, detecting levels of cardiolipin isoforms comprises chromatography, mass spectrometry, ELISA, Western blotting, immunodetection, or immunoprecipitation.

In some embodiments, assessing cardiolipin remodeling comprises a mitochondrial function assay.

In some embodiments, the mitochondrial function assay comprises the use of peripheral blood mononuclear cells (PBMCs), leukocytes, or ex vivo tissues.

In some embodiments, the method further comprises administering to the subject a therapeutically effective amount of the peptide D-Arg-2′6′-Dmt-Lys-Phe-NH2or a pharmaceutically acceptable salt thereof.

In some embodiments, the peptide is administered daily for 6 weeks or more.

In some embodiments, the peptide is administered daily for 12 weeks or more.

In some embodiments, the mitochondrial dysfunction results from hypertension; ischemic heart disease; exposure to a cardiotoxic compound; myocarditis; thyroid disease; viral infection; gingivitis; drug abuse; alcohol abuse; pericarditis; atherosclerosis; vascular disease; hypertrophic cardiomyopathy; acute myocardial infarction; left ventricular systolic dysfunction; coronary bypass surgery; starvation; an eating disorder; or a genetic defect.

In some embodiments, the subject is human.

In some embodiments, the peptide is administered orally, topically, systemically, intravenously, subcutaneously, intraperitoneally, or intramuscularly

In some embodiments, the method further comprises separately, sequentially or simultaneously administering a cardiovascular agent to the subject.

In some embodiments, the cardiovascular agent is selected from the group consisting of: an anti-arrhythmia agent, a vasodilator, an anti-anginal agent, a corticosteroid, a cardioglycosidc, a diuretic, a sedative, an angiotensin converting enzyme (ACE) inhibitor, an angiotensin II antagonist, a thrombolytic agent, a calcium channel blocker, a throboxane receptor antagonist, a radical scavenger, an anti-platelet drug, a β-adrenaline receptor blocking drug, α-receptor blocking drug, a sympathetic nerve inhibitor, a digitalis formulation, an inotrope, and an antihyperlipidemic drug.

In some embodiments, the pharmaceutically acceptable salt comprises acetate or trifluoroacetate salt.

In some embodiments, the biological sample comprises tissue, a cell, or a mitochondria from the subject.

In one aspect, the present disclosure provides a method for assessing cardiolipin content and composition in a mammalian subject in need thereof, the method comprising assessing cardiolipin remodeling in a biological sample from the subject.

In some embodiments, assessing cardiolipin remodeling comprises detecting levels of cardiolipin remodeling enzymes.

In some embodiments, detecting levels of cardiolipin remodeling enzymes comprises detecting the level of one or more of TAZ1, MLCL AT1, or ALCAT1 mRNA compared to a normal control subject.

In some embodiments, the level of MLCL AT1 or ALCAT1 mRNA is elevated about 2.5-fold compared to a normal control subject.

In some embodiments, the level of TAZ1 mRNA is reduced about 2.5-fold compared to a normal control subject.

In some embodiments, assessing cardiolipin remodeling comprises detecting levels of cardiolipin isoforms compared to a normal control subject.

In some embodiments, detecting one or more of TAZ1, MLCL AT1, or ALCAT1 mRNA comprises RT-PCR, in situ hybridization, or Northern blotting.

In some embodiments, detecting levels of cardiolipin isoforms comprises chromatography, mass spectrometry, ELISA, Western blotting, immunodetection, or immunoprecipitation.

In some embodiments, assessing cardiolipin remodeling comprises a mitochondrial function assay.

In some embodiments, the mitochondrial function assay comprises the use of peripheral blood mononuclear cells (PBMCs), leukocytes, or ex vivo tissues.

In some embodiments, the method further comprises administering to the subject a therapeutically effective amount of the peptide D-Arg-2′6′-Dmt-Lys-Phe-NH2or a pharmaceutically acceptable salt thereof.

In some embodiments, the peptide is administered daily for 6 weeks or more.

In some embodiments, the peptide is administered daily for 12 weeks or more.

In some embodiments, the cardiolipin content and composition is aberrant due to hypertension; ischemic heart disease; exposure to a cardiotoxic compound; myocarditis; thyroid disease; viral infection; gingivitis; drug abuse; alcohol abuse; pericarditis; atherosclerosis; vascular disease; hypertrophic cardiomyopathy; acute myocardial infarction; left ventricular systolic dysfunction; coronary bypass surgery; starvation; an eating disorder; or a genetic defect.

In some embodiments, the subject is human.

In some embodiments, the peptide is administered orally, topically, systemically, intravenously, subcutaneously, intraperitoneally, or intramuscularly

In some embodiments, the method further comprises separately, sequentially or simultaneously administering a cardiovascular agent to the subject.

In some embodiments, the cardiovascular agent is selected from the group consisting of: an anti-arrhythmia agent, a vasodilator, an anti-anginal agent, a corticosteroid, a cardioglycoside, a diuretic, a sedative, an angiotensin converting enzyme (ACE) inhibitor, an angiotensin II antagonist, a thrombolytic agent, a calcium channel blocker, a throboxane receptor antagonist, a radical scavenger, an anti-platelet drug, a β-adrenaline receptor blocking drug, α-receptor blocking drug, a sympathetic nerve inhibitor, a digitalis formulation, an inotrope, and an antihyperlipidemic drug.

In some embodiments, the pharmaceutically acceptable salt comprises acetate or trifluoroacetate salt.

In some embodiments, the biological sample comprises tissue, a cell, or a mitochondria from the subject.

