Theranostics for hypertension-induced myocardial microbleeds

Abstract

The present invention relates to cardiovascular imaging methods and / or blood marker measurement methods for detecting, diagnosing, and / or prognosing cardiac or myocardial microhemorrhages, particularly in subjects with hypertension and cardiovascular disease. Treatment methods are also provided for subjects identified, diagnosed, prognosed, or detected as having cardiac or myocardial microhemorrhages. In some embodiments, the subject has hypertension-induced microhemorrhages but has not previously had a myocardial infarction or undergone reperfusion therapy. TIFF2022554269000002.tif107132

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 62 / 928,717, filed October 31, 2019, which is incorporated herein by reference in its entirety.

[0002] FIELD OF THE INVENTION The present invention relates to cardiovascular imaging methods for the detection, diagnosis, and / or prognosis of various conditions associated with hypertension and cardiovascular disease, as well as treatment thereof. [Background technology]

[0003] background All publications cited herein are incorporated by reference in their entirety to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. The following description contains information that may be useful in understanding the present invention. None of the information provided herein is admitted to be prior art to or relevant to the invention claimed herein, or that any publication specifically or implicitly mentioned is prior art.

[0004] Many people suffer from or are at risk of developing hypertension. Hypertension can lead to a variety of adverse outcomes, including cardiovascular disease, cardiac arrhythmias, sudden cardiac death, and death. Thus, there is a need for methods for detecting, diagnosing, and / or treating symptoms associated with hypertension to reduce adverse outcomes in people with hypertension, particularly those who have hypertension and are at risk of developing or suffering from cardiovascular disease or heart-related conditions. Summary of the Invention

[0005] The following embodiments and aspects thereof are described and illustrated in conjunction with compositions and methods that are intended to be representative and exemplary, not limiting in scope.

[0006] In various embodiments, a method for identifying and / or assessing at least one microhemorrhage in a subject's heart is provided, the method comprising obtaining at least one image of the subject's heart and detecting at least one iron deposit in the subject's heart from the at least one image, wherein the detection of the iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart. In other embodiments, a method for identifying and / or assessing at least one microhemorrhage in a subject's heart is provided, the method comprising detecting and / or measuring the presence of at least one iron deposit in the subject's heart by measuring a level of a hemoglobin breakdown product or a level of a ferroptosis marker from the subject's blood, wherein the detection of the iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart.

[0007] In various embodiments, at least one microhemorrhage in the heart is caused by or associated with hypertension. In some embodiments, the subject has hypertension. In some embodiments, the subject has been diagnosed with hypertension. In some embodiments, the subject has secondary hypertension caused by another condition, and thus the subject also has or has been diagnosed with chronic kidney disease, polycystic kidney disease, aortic coarctation of the heart, adrenal gland disorders, hyperthyroidism, renal artery stenosis, sleep disorders, or pheochromocytoma. In some embodiments, the subject has taken or is taking blood pressure medication. In some embodiments, the subject has stress-induced cardiomyopathy. In some embodiments, the subject has not had a myocardial infarction or reperfusion therapy-induced hemorrhage, and in some further aspects, the subject has not had a hemorrhagic myocardial infarction, and in some further aspects, the subject has not undergone reperfusion therapy and therefore does not have reperfusion hemorrhage. In some embodiments, the subject has not had a myocardial infarction or undergone reperfusion therapy within the past 3 days, 7 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 2 years, 3 years, or 5 years. In some embodiments, a subject in need of one or more methods disclosed herein has any one or more of the characteristics described above, and in some embodiments, a subject in need thereof desires identification, detection, prognosis, or diagnosis of myocardial microhemorrhage.

[0008] In various embodiments, a method is provided for detecting at least one microhemorrhage in the heart of a subject, the method comprising detecting and / or measuring the presence of at least one iron deposit in the heart of the subject by performing cardiac imaging and / or measuring a level of a hemoglobin breakdown product or a level of a ferroptosis marker from the subject's blood, wherein the detection of the at least one iron deposit in the heart of the subject is indicative of and / or is the detection of at least one microhemorrhage in the heart of the subject.

[0009] In various embodiments, a method for prognosing at least one microhemorrhage in a subject's heart is provided, the method comprising detecting and / or measuring the presence of at least one iron deposit in the subject's heart by performing cardiac imaging and / or measuring a level of a hemoglobin breakdown product or a level of a ferroptosis marker from the subject's blood, wherein detection of the at least one iron deposit in the subject's heart is prognostic of at least one microhemorrhage in the subject's heart.

[0010] In various embodiments, a method is provided for diagnosing at least one microhemorrhage in a subject's heart, the method comprising detecting and / or measuring the presence of at least one iron deposit in the subject's heart by performing cardiac imaging and / or measuring a level of a hemoglobin breakdown product or a level of a ferroptosis marker from the subject's blood, wherein detection of at least one iron deposit in the subject's heart is diagnostic of at least one microhemorrhage in the subject's heart.

[0011] In various embodiments, methods are provided for assessing and / or determining a subject's risk of developing at least one microhemorrhage in the heart, the methods comprising detecting and / or measuring the presence of at least one iron deposit in the subject's heart by performing cardiac imaging and / or measuring levels of hemoglobin breakdown products or levels of ferroptosis markers from the subject's blood, wherein detection of at least one iron deposit in the subject's heart indicates an increased risk of developing at least one microhemorrhage in the subject's heart.

[0012] In various embodiments, a method for identifying a subject with cardiac microhemorrhage is provided, the subject having hypertension, the method comprising: selecting a subject diagnosed with or suffering from hypertension; and performing a clinical imaging scan of at least a portion of the subject's heart to detect iron-containing deposits in the portion of the heart.Representative clinical imaging scans include single-photon emission computed tomography (SPECT), positron emission tomography (PET), computed tomography (CT), ultrasound (US), magnetic resonance imaging (MRI), and combinations thereof.One or more of these imaging techniques can tag the degradation products of hemoglobin or proteins associated with ferroptosis.

[0013] In various embodiments, a method is provided for identifying and / or assessing at least one microhemorrhage in at least one portion of a subject's heart, the method comprising detecting at least one iron deposit in at least one portion of the subject's heart by performing clinical imaging of the at least one portion of the heart, wherein detection of the at least one iron deposit in the at least one portion of the subject's heart is indicative of at least one microhemorrhage in the at least one portion of the subject's heart.

[0014] In various embodiments, a method is provided for detecting at least one microhemorrhage in at least one portion of a subject's heart, the method comprising detecting at least one iron deposit in at least one portion of the subject's heart by performing clinical imaging of the at least one portion of the heart, wherein the detection of the at least one iron deposit in the at least one portion of the subject's heart is indicative of and / or is the detection of at least one microhemorrhage in the at least one portion of the subject's heart.

[0015] In various embodiments, a method for prognosing at least one microhemorrhage in at least one portion of a subject's heart is provided, the method comprising detecting at least one iron deposit in at least one portion of the subject's heart by performing clinical imaging of the at least one portion of the heart, wherein the detection of the at least one iron deposit in the at least one portion of the subject's heart is prognostic of at least one microhemorrhage in the at least one portion of the subject's heart.

[0016] In various embodiments, a method is provided for diagnosing at least one microhemorrhage in at least one portion of a subject's heart, the method comprising detecting at least one iron deposit in at least one portion of the subject's heart by performing clinical imaging of the at least one portion of the heart, wherein the detection of the at least one iron deposit in the at least one portion of the subject's heart is diagnostic of at least one microhemorrhage in the at least one portion of the subject's heart.

[0017] In various embodiments, a method is provided for assessing and / or determining a subject's risk of developing at least one microhemorrhage in at least one portion of the heart, the method comprising detecting at least one iron deposit in at least one portion of the subject's heart by performing clinical imaging of the at least one portion of the heart, wherein detection of the at least one iron deposit in the at least one portion of the subject's heart is indicative of an increased risk of the subject for developing at least one microhemorrhage in the at least one portion of the subject's heart.

[0018] In various embodiments, a method is provided for identifying and / or assessing at least one microhemorrhage in at least one cardiac tissue of a subject, the method comprising detecting at least one iron deposit in the at least one cardiac tissue of the subject by performing clinical imaging of the at least one cardiac tissue, wherein the detection of the at least one iron deposit in the at least one cardiac tissue is indicative of at least one microhemorrhage in the at least one cardiac tissue of the subject.

[0019] In various embodiments, a method is provided for detecting at least one microhemorrhage in at least one cardiac tissue of a subject, the method comprising detecting at least one iron deposit in at least one cardiac tissue of the subject by performing clinical imaging of the at least one cardiac tissue, wherein the detection of the at least one iron deposit in the at least one cardiac tissue is indicative of and / or is the detection of at least one microhemorrhage in the at least one cardiac tissue of the subject.

[0020] In various embodiments, a method for prognosing at least one microhemorrhage in at least one cardiac tissue of a subject is provided, the method comprising detecting at least one iron deposit in at least one cardiac tissue of the subject by performing clinical imaging of the at least one cardiac tissue, wherein the detection of the at least one iron deposit in the heart is prognostic of at least one microhemorrhage in the subject's heart.

[0021] In various embodiments, a method is provided for diagnosing at least one microhemorrhage in at least one cardiac tissue of a subject, the method comprising detecting at least one iron deposit in at least one cardiac tissue of the subject by performing clinical imaging of the at least one cardiac tissue, wherein the detection of the at least one iron deposit in the at least one cardiac tissue is diagnostic of at least one microhemorrhage in the at least one cardiac tissue of the subject.

[0022] In various embodiments, a method is provided for assessing and / or determining a subject's risk of developing at least one microhemorrhage in at least one cardiac tissue, the method comprising detecting at least one iron deposit in the subject's at least one cardiac tissue by performing clinical imaging of the at least one cardiac tissue, wherein detection of the at least one iron deposit in the at least one cardiac tissue is indicative of the subject's increased risk for developing at least one microhemorrhage in the subject's at least one cardiac tissue.

[0023] In various embodiments, a method is provided for identifying and / or assessing at least one microhemorrhage in a subject's heart, the method comprising detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart and comparing the image of the subject's heart with at least one image from a reference sample, wherein the presence of the at least one iron deposit in the subject's heart compared to the reference sample is indicative of at least one microhemorrhage in the subject's heart.

[0024] In various embodiments, a method for detecting at least one microhemorrhage in a subject's heart is provided, the method comprising detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart and comparing the image of the subject's heart with at least one image from a reference sample, wherein the presence of the at least one iron deposit in the subject's heart compared to the reference sample is indicative of at least one microhemorrhage in the subject's heart.

[0025] In various embodiments, a method for prognosing at least one microhemorrhage in a subject's heart is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart; and comparing the image of the subject's heart with at least one image from a reference sample, wherein the presence of the at least one iron deposit in the subject's heart compared to the reference sample is prognostic of at least one microhemorrhage in the subject's heart.

[0026] In various embodiments, a method is provided for diagnosing at least one microhemorrhage in a subject's heart, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart; and comparing the image of the subject's heart with at least one image from a reference sample, wherein the presence of the at least one iron deposit in the subject's heart compared to the reference sample is diagnostic of at least one microhemorrhage in the subject's heart.

[0027] In various embodiments, a method is provided for assessing and / or determining a subject's risk of developing at least one microhemorrhage in the heart, the method comprising detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart and comparing the image of the subject's heart with at least one image from a reference sample, wherein the presence of the at least one iron deposit in the subject's heart compared to the reference sample indicates an increased risk of the subject for developing at least one microhemorrhage in the subject's heart.

[0028] In various embodiments, a method is provided for determining the progression of at least one microhemorrhage in a subject's heart, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart; and comparing the image of the subject's heart with at least one image from a reference sample, wherein a change in the image of the subject's heart compared to the at least one image from the reference sample indicates the progression of at least one microhemorrhage in the subject's heart.

[0029] In various embodiments, a method for identifying and / or assessing cardiovascular disease in a subject is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical cardiac imaging, wherein the detection of the iron deposit in the subject's heart is indicative of the presence of at least one microhemorrhage in the subject's heart; and comparing the image of the subject's heart with at least one image from a reference sample, wherein a change in the image of the subject's heart compared to the at least one image from the reference sample is indicative of cardiovascular disease in the subject.

[0030] In various embodiments, a method for detecting cardiovascular disease in a subject is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart, wherein the detection of the at least one iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the at least one image of the subject's heart with at least one image from a reference sample, wherein a change in the at least one image of the subject's heart compared to the at least one image from the reference sample is indicative of cardiovascular disease in the subject.

[0031] In various embodiments, a method for detecting cardiovascular disease in a subject is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart, wherein the detection of the at least one iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the at least one image of the subject's heart with at least one image from a reference sample, wherein a change in the at least one image of the subject's heart compared to the at least one image from the reference sample is indicative of cardiovascular disease in the subject.

[0032] In various embodiments, a method for prognosing cardiovascular disease in a subject is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical cardiac imaging, wherein the detection of the iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the image of the subject's heart with at least one image from a reference sample, wherein a change in the image of the subject's heart compared to the at least one image from the reference sample is prognostic of cardiovascular disease in the subject.

[0033] In various embodiments, a method for diagnosing cardiovascular disease in a subject is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart, wherein the detection of the iron deposit in the subject's heart is indicative of the presence of at least one microhemorrhage in the subject's heart; and comparing the image of the subject's heart with at least one image from a reference sample, wherein a change in the image of the subject's heart compared to the at least one image from the reference sample is diagnostic of cardiovascular disease in the subject.

[0034] In various embodiments, a method for assessing and / or determining a risk of developing cardiovascular disease in a subject is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical cardiac imaging, wherein the detection of the iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the image of the subject's heart with at least one image from a reference sample, wherein a change in the image of the subject's heart compared to the at least one image from the reference sample indicates an increased risk of the subject for developing cardiovascular disease in the subject.

[0035] In various embodiments, a method for determining the progression of cardiovascular disease in a subject is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart, wherein the detection of the iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the image of the subject's heart with at least one image from a reference sample, wherein a change in the image of the subject's heart compared to the at least one image from the reference sample indicates the progression of cardiovascular disease in the subject.

[0036] In various embodiments, the cardiovascular disease can be one, two, or multiple cardiac conditions.

[0037] In various embodiments, a method is provided for identifying at least one biomarker for cardiovascular disease in a subject, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart; correlating the at least one iron deposit in the at least one image of the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject with a biomarker signature from a reference sample, wherein the at least one biomarker for cardiovascular disease is identified.

[0038] In various embodiments, a method for identifying and / or assessing cardiovascular disease in a subject is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject with a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is indicative of cardiovascular disease in the subject.

[0039] In various embodiments, a method for detecting cardiovascular disease in a subject is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject with a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is indicative of and / or detection of cardiovascular disease in the subject.

[0040] In various embodiments, a method for prognosing cardiovascular disease in a subject is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject with a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is prognostic of cardiovascular disease in the subject.

[0041] In various embodiments, a method for diagnosing cardiovascular disease in a subject is provided, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject with a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is diagnostic of cardiovascular disease in the subject.

[0042] In various embodiments, a method is provided for determining the progression of cardiovascular disease in a subject, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject with a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is indicative of the progression of cardiovascular disease in the subject.

[0043] In some embodiments, there is provided a method for assessing and / or determining a risk of developing cardiovascular disease in a subject, the method comprising: detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject with a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is indicative of an increased risk of the subject for developing cardiovascular disease.

[0044] In various embodiments, a method is provided for assessing and / or determining the risk of at least one cardiac arrhythmia in a subject, the method comprising detecting at least one iron deposit in the subject's heart by performing clinical cardiac imaging, wherein the presence of the at least one iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and wherein the at least one microhemorrhage in the subject's heart is indicative of an increased risk of at least one cardiac arrhythmia in the subject.

[0045] In various embodiments, a method is provided for assessing and / or determining risk of sudden cardiac death in a subject, the method comprising detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart, wherein the presence of the at least one iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and wherein the at least one microhemorrhage in the subject's heart is indicative of an increased risk of sudden cardiac death in the subject.

[0046] In various embodiments, a method for assessing and / or determining risk of cardiovascular disease in a subject is provided, the method comprising detecting at least one iron deposit in the subject's heart by performing clinical imaging of the heart, wherein the presence of the at least one iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and wherein the at least one microhemorrhage in the subject's heart is indicative of an increased risk of cardiovascular disease in the subject.

[0047] In some embodiments, the subject is a human. In some embodiments, the subject has at least one selected from the group consisting of hypertension, hypertensive emergency, acute hypertensive emergency, persistent hypertension, and combinations thereof. In some embodiments, the subject has cardiomyopathy, such as stress-induced cardiomyopathy.

[0048] Various embodiments provide a method for treating or reducing the severity or risk of cardiovascular disease associated with myocardial microhemorrhage in a subject with hypertension or a hypertensive emergency, the method comprising administering to the subject an effective amount of a composition comprising an iron chelator, a heme-binding protein, a heme-degrading protein, a heme-blocking protein, or an agent effective to increase the amount of a heme-binding protein or a heme-degrading protein.

[0049] Further embodiments of these methods of treatment include selecting a subject having iron deposits in at least one portion of the heart and administering to the subject an effective amount of the composition. In some aspects, the subject having cardiac iron deposits is selected and / or identified by performing a clinical imaging scan and / or measuring blood-based markers to detect the presence of iron in the heart.

[0050] In some embodiments, the method further comprises treating the subject with a therapeutic agent and / or selecting a therapeutic agent for the subject and / or providing a therapeutic agent to the subject and / or administering a therapeutic agent to the subject. In some embodiments, the therapeutic agent is at least one chelating agent. In some embodiments, the chelating agent is selected from the group consisting of deferoxamine, deferasirox, deferiprone, hinokitiol, dexrazoxane, 2,2'-bipyridyl, and combinations thereof. In some embodiments, the chelating agent is an iron chelating agent. In some embodiments, the iron chelating agent is selected from the group consisting of an intracellular iron chelator, an extracellular iron chelator, or a combination thereof. In some embodiments, the intracellular iron chelator chelates ferrous iron, ferric iron, and combinations thereof. In some embodiments, the extracellular iron chelator chelates ferrous iron, ferric iron, and combinations thereof. In some embodiments, the therapeutic agent is selected from the group consisting of at least one heme-binding protein, at least one heme-degrading protein, at least one heme-blocking agent / protein, and combinations thereof. In some embodiments, the method further comprises administering to the subject an amount of an agent effective to increase the amount of at least one selected from the group consisting of at least one heme-binding protein, at least one heme-degrading protein, and combinations thereof in the subject. In some embodiments, the heme-binding protein is selected from the group consisting of hemopexin, haptoglobin, albumin, ferritin, alpha1-microglobulin, alpha1-antitrypsin, glutathione-S-transferase, heme-binding protein / liver fatty acid-binding protein, heme-binding protein 23 / peroxiredoxin, p22 heme-binding protein, and glyceraldehyde-3-phosphate dehydrogenase, and combinations thereof. In some embodiments, the heme-degrading protein is selected from the group consisting of heme oxygenase 1, nuclear factor E2-related factor 2 (Nrf2), and combinations thereof. In some embodiments, the heme blocking agent / protein is hepcidin.In some embodiments, the factor is selected from FLVCR1a (feline leukemia virus subgroup C receptor 1a), ABCG2 (ATP-binding cassette subfamily G member 2), FLVCR2, and combinations thereof. In some embodiments, the heme-binding protein is a heme scavenger.

[0051] In some embodiments, the iron deposits (also known as iron-containing deposits) comprise iron oxide. In some embodiments, the iron deposits comprise at least one compound comprising iron. In some embodiments, the compound is selected from the group consisting of heme, heme derivatives, and combinations thereof. In some embodiments, the iron is in any possible oxidation state. In some embodiments, the oxidation state is selected from +2, +2 / +3, +3, +4, and combinations thereof. In some embodiments, the iron is selected from the group consisting of ferrous iron, ferric iron, and combinations thereof. In some embodiments, the iron is in any form.

[0052] In some embodiments, the portion of the heart is selected from the group consisting of epicardium, myocardium, endocardium, valve tissue, and combinations thereof. In some embodiments, the portion of the heart is selected from the group consisting of valve tissue, endocardium, pulmonary valve, brachiocephalic trunk, superior vena cava, right pulmonary artery, right pulmonary vein, tricuspid valve, right ventricle, inferior vena cava, descending aorta, papillary muscle, interventricular septum, epicardium, left ventricle, myocardium, mitral valve, aortic valve, left pulmonary vein, left pulmonary artery, aorta, left subclavian artery, pulmonary trunk, left common carotid artery, and combinations thereof. In some embodiments, the portion of the heart is or includes myocardium.

[0053] In some embodiments, the change is in at least one iron deposit, hi some embodiments, the change is in at least one selected from the group consisting of amount, quantity, concentration, shape, size, and pattern of the at least one iron deposit.

[0054] In some embodiments, the reference sample is the heart from a control subject, and the control subject does not have at least one iron deposit in the heart.In some embodiments, the reference sample is the heart from a control subject, and the control subject does not have at least one microhemorrhage in the heart.In some embodiments, the reference sample is the heart from a subject at an earlier time point.

[0055] In some embodiments, the cardiovascular disease is selected from the group consisting of infarcted myocardium, coronary artery disease, coronary heart disease, ischemic heart disease, cardiomyopathy, stroke, hypertensive heart disease, heart failure, pulmonary heart disease, ischemic syndrome, coronary microvascular disease, cardiac arrhythmia, rheumatic heart disease, aortic aneurysm, cardiomyopathy, atrial fibrillation, congenital heart disease, endocarditis, inflammatory heart disease, endocarditis, inflammatory cardiac hypertrophy, myocarditis, valvular heart disease, cerebrovascular disease, peripheral arterial disease, acute stress-induced cardiomyopathy (takotsubo cardiomyopathy), diffuse myocardial fibrosis, and combinations thereof. In some embodiments, the cardiovascular disease comprises sudden heart failure or death, myocardial electrical or mechanical abnormalities, myocardial inflammation, ventricular arrhythmia, atrial fibrillation, myocardial fibrosis, and / or adverse cardiac remodeling. In some embodiments, the cardiovascular disease comprises myocardial fibrosis. In some embodiments, the cardiovascular disease comprises myocardial fibrosis. In some embodiments, the cardiovascular disease consists essentially of myocardial fibrosis.

[0056] In some embodiments, the reference sample is a heart from a subject before the subject is treated for cardiovascular disease, hi some embodiments, the reference sample is a heart from a subject who has been successfully treated for cardiovascular disease.

[0057] In some embodiments, the reference sample is a heart from a control subject, and the control subject does not have cardiovascular disease. In some embodiments, the control subject does not have hypertension, hypertensive emergency, acute hypertensive emergency, or persistent hypertension. In some embodiments, the reference sample is a heart from a control subject, and the control subject has cardiovascular disease.

[0058] In some embodiments, the reference sample is from the heart of a subject who has been successfully treated for hypertension, hi some embodiments, the reference sample is from the heart of the subject at an earlier time point.

[0059] In some embodiments, obtaining at least one clinical image or performing at least one clinical imaging comprises operating a clinical imaging machine / scanner to obtain at least one clinical image. In some embodiments, obtaining at least one CT image comprises operating a CT imaging machine / scanner to obtain at least one CT image. In some embodiments, obtaining at least one SPECT image comprises operating a SPECT imaging machine / scanner to obtain at least one SPECT image. In some embodiments, obtaining at least one PET image comprises operating a PET imaging machine / scanner to obtain at least one PET image. In some embodiments, obtaining at least one magnetic resonance image comprises operating a magnetic resonance imaging machine / scanner to obtain at least one magnetic resonance image.

[0060] In some embodiments, the cardiac tissue is selected from the group consisting of valve tissue, endocardium, pulmonary valve, brachiocephalic artery, superior vena cava, right pulmonary artery, right pulmonary vein, tricuspid valve, right ventricle, inferior vena cava, descending aorta, papillary muscle, interventricular septum, epicardium, left ventricle, myocardium, mitral valve, aortic valve, left pulmonary vein, left pulmonary artery, aorta, left subclavian artery, pulmonary trunk, left common carotid artery, and combinations thereof. [The present invention 1001] 1. A method for identifying, detecting, prognosing, and / or diagnosing at least one microhemorrhage in the heart of a subject in need thereof, comprising: detecting and / or measuring the presence of at least one iron deposit in the heart of the subject by performing imaging of the heart and / or measuring the level of a hemoglobin breakdown product or a level of a ferroptosis marker from the blood of the subject. Including, the presence of said iron deposits in the heart of said subject is indicative of at least one microhemorrhage in the heart of said subject. The method. [The present invention 1002] 1001. The method of claim 1001, wherein said subject has been diagnosed with hypertension, a hypertensive emergency, an acute hypertensive emergency, persistent hypertension, or a combination thereof. [The present invention 1003] The method of claim 1001, wherein said detecting and / or measuring comprises performing imaging of the subject's heart, said imaging being selected from the group consisting of single photon emission computed tomography (SPECT), positron emission tomography (PET), computed tomography (CT), ultrasound (US), magnetic resonance imaging (MRI), and combinations thereof. [The present invention 1004] 1004. The method of claim 1003, wherein said detecting and / or measuring comprises performing a SPECT scan, a PET scan, or both, of said subject's heart. [The present invention 1005] The method of claim 1001, wherein the step of detecting and / or measuring the presence of at least one iron deposit comprises the level of a hemoglobin degradation product or the level of a ferroptosis marker from the subject's blood, wherein the hemoglobin degradation product is selected from the group consisting of iron, bilirubin, globin, and combinations thereof, and the ferroptosis marker comprises glutathione peroxidase 4 (GPX4). [The present invention 1006] comparing the image of the subject's heart, the level of the hemoglobin breakdown product, or the level of the ferroptosis marker with that obtained from a reference sample. The method of the present invention 1001 further comprising: [The present invention 1007] 1006. The method of claim 1006, wherein the presence of said at least one iron deposit in the heart of said subject compared to said reference sample is indicative of at least one microhemorrhage in the heart of said subject, or is prognostic or diagnostic of said microhemorrhage. [The present invention 1008] a change in the image of the subject's heart, the level of the hemoglobin breakdown product, or the level of the ferroptosis marker compared to that obtained from the reference sample indicates the development of at least one microhemorrhage in the subject's heart. 1006. The method of claim 1006, for determining the progression of at least one microhemorrhage in the heart of said subject. [The present invention 1009] 1. A method for identifying, detecting, prognosing, diagnosing, and / or determining the progression of cardiovascular disease in a subject in need thereof, comprising: detecting at least one iron deposit in the subject's heart by performing cardiac imaging and / or measuring a level of a hemoglobin breakdown product or a level of a ferroptosis marker from the subject's blood, wherein the detection of the iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; comparing the image of the subject's heart, the level of the hemoglobin breakdown product, or the level of the ferroptosis marker with that obtained from a reference sample, wherein a change in the image of the subject's heart, the level of the hemoglobin breakdown product, or the level of the ferroptosis marker compared to that obtained from the reference sample is indicative of cardiovascular disease in the subject, or is prognostic or diagnostic of the cardiovascular disease, or is indicative of the progression of the cardiovascular disease; The method comprising: [The present invention 1010] correlating the at least one iron deposit in the at least one image of the subject's heart, or the at least one iron deposit indicated by the level of the hemoglobin breakdown product or the level of the ferroptosis marker in the subject, with at least one microhemorrhage in the subject's heart, to obtain a biomarker signature from at least one MR image of the subject. further comprising the comparing step compares the biomarker signature obtained from the image of the subject with a biomarker signature from a reference sample, thereby identifying at least one biomarker for cardiovascular disease; a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is indicative of the cardiovascular disease in the subject, or is detection, prognosis, or diagnosis of the cardiovascular disease, or is indicative of the progression of the cardiovascular disease. The method of the present invention 1009. [The present invention 1011] 1009. The method of claim 1009, wherein said subject has been diagnosed with hypertension, a hypertensive emergency, an acute hypertensive emergency, persistent hypertension, or a combination thereof. [The present invention 1012] 1009. The method of claim 1009, wherein said cardiovascular disease comprises cardiomyopathy, myocardial fibrosis, cardiac arrhythmia, or sudden cardiac death. [The present invention 1013] 1. A method of treating a subject experiencing, recovering from, and / or at risk of developing cardiovascular disease associated with myocardial microhemorrhage, comprising: administering an effective amount of the composition to said subject. Including, the composition comprises a chelating agent, a heme-binding protein, a heme-degrading protein, a heme-blocking agent, an agent effective to increase the amount of a heme-binding protein or a heme-degrading protein, or a combination thereof; The method. [The present invention 1014] The method of claim 1013, wherein said subject has hypertension, a hypertensive emergency, an acute hypertensive emergency, chronic hypertension, or a combination thereof. [The present invention 1015] Detecting and / or measuring the presence of at least one iron deposit in the heart of said subject, thereby linking cardiovascular disease to myocardial microhemorrhage. further comprising The detecting and / or measuring comprises performing imaging of the heart and / or measuring the level of a hemoglobin degradation product or a ferroptosis marker from the subject's blood. The method of the present invention 1013. [The present invention 1016] the chelating agent is selected from the group consisting of deferoxamine, deferasirox, deferiprone, hinokitiol, dexrazoxane, 2,2'-bipyridyl, and combinations thereof; the heme-binding protein is selected from the group consisting of hemopexin, haptoglobin, albumin, ferritin, alpha 1-microglobulin, alpha 1-antitrypsin, glutathione-S-transferase, heme-binding protein / liver fatty acid-binding protein, heme-binding protein 23 / peroxiredoxin, p22 heme-binding protein, and glyceraldehyde-3-phosphate dehydrogenase, and combinations thereof; the heme-degrading protein is selected from the group consisting of heme oxygenase 1, nuclear factor E2-related factor 2 (Nrf2), and combinations thereof; the heme blocker comprises hepcidin, and / or The factor is selected from feline leukemia virus subgroup C receptor (FLVCR) 1a (FLVCR1a), FLVCR2, ATP-binding cassette subfamily G member 2 (ABCG2), and combinations thereof. The method of the present invention 1013. [The present invention 1017] imaging the subject's heart and / or measuring levels of hemoglobin breakdown products or levels of ferroptosis markers from the subject's blood after administering the composition to detect levels of iron deposition in the myocardium or other parts of the heart. The method of the present invention 1013 further comprises: [The present invention 1018] administering a subsequent dose of the composition to the subject if the myocardium or other portion of the heart exhibits the same or higher level of iron deposition compared to the level before the subject was administered the previous dose, or discontinuing administration of the composition if there are no iron deposits in the myocardium or other portion of the heart. The method of the present invention 1017 further comprising: [The present invention 1019] The method of claim 1013, wherein the subject is a human. [The present invention 1020] The method of claim 1013, wherein the subject has not had a myocardial infarction or reperfusion therapy-induced hemorrhage, or the subject has not had a myocardial infarction or reperfusion therapy-induced hemorrhage in the past 3 days, 7 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 2 years, 3 years, or 5 years. [The present invention 1021] The method of any of claims 1001 to 1008, for identifying at least one microhemorrhage in the heart of said subject. [The present invention 1022] The method of any of claims 1001 to 1008, for detecting at least one microhemorrhage in the heart of said subject. [The present invention 1023] The method of any of claims 1001 to 1008, which is for prognosticating at least one microbleed in the heart of said subject. [The present invention 1024] The method of any of claims 1001 to 1008, which is for diagnosing at least one microbleed in the heart of said subject. [The present invention 1025] The method of any of claims 1009 to 1012, which is for identifying said cardiovascular disease in said subject, or for identifying a subject having said cardiovascular disease. [The present invention 1026] The method of any one of claims 1009 to 1012, which is for detecting said cardiovascular disease in said subject. [The present invention 1027] The method of any of claims 1009 to 1012, which is for predicting the prognosis of said cardiovascular disease in said subject. [The present invention 1028] The method of any of claims 1009 to 1012, which is for determining the progression of said cardiovascular disease in said subject. [Brief explanation of the drawings]

[0061] Representative embodiments are illustrated in the referenced figures. It is intended that the embodiments and figures disclosed herein be considered illustrative rather than restrictive.

[0062] [Figure 1] A is a representative Prussian blue stained image illustrating iron in myocardial tissue, and B is a representative Masson's trichrome stained image of elastin illustrating the resulting myocardial fibrosis, both from wild-type rats treated with angiotensin II to induce hypertension. DETAILED DESCRIPTION OF THE INVENTION

[0063] Detailed Description of the Invention All references cited herein are incorporated by reference in their entirety as if fully set forth. Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Allen et al., Remington: The Science and Practice of Pharmacy, 22nd ed., Pharmaceutical Press (September 15, 2012); Hornyak et al., Introduction to Nanoscience and Nanotechnology, CRC Press (2008); Smith, March's Advanced Organic Chemistry Reactions, Mechanisms and Structure, 7th ed., J. Wiley & Sons (New York, NY 2013); Singleton, Dictionary of DNA and Genome Technology, 3rd ed., Wiley-Blackwell (November 28, 2012); and Green and Sambrook, Molecular Cloning: A Laboratory Manual, 4th ed., Cold Spring Harbor Laboratory Press (Cold Spring Harbor, NY 2012) will provide those of skill in the art with a general guide to many of the terms used herein.

[0064] Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. Other features and advantages of the present invention will become apparent from the following detailed description, taken in conjunction with the accompanying drawings, which illustrate, by way of example, various features of embodiments of the present invention. Indeed, the present invention is in no way limited to the methods and materials described. For convenience, certain terms used herein in the specification, examples, and appended claims are collected here.

[0065] Unless otherwise specified or implicit from context, the following terms and phrases include the meanings provided below. Unless expressly specified otherwise or apparent from context, the following terms and phrases do not exclude the meaning that such term or phrase has acquired in the relevant art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It is to be understood that the invention is not limited to the particular methodology, protocols, and reagents, etc., described herein, as such may vary. Since the scope of the present invention is limited only by the claims, the definitions and terminology used herein are provided to aid in describing particular embodiments and are not intended to limit the claimed invention.

[0066] The terms "comprising" or "comprise" are used in reference to compositions, methods, systems, articles of manufacture, and their respective component(s) that are useful for an embodiment, but are open to including unspecified elements, whether or not still useful. In general, those of skill in the art will understand that the terms used herein are generally intended as "open-ended" terms (e.g., the term "including" should be interpreted as "including but not limited to," the term "having" should be interpreted as "having at least," the term "include" should be interpreted as "include but not limited to," etc.). Although the open-ended term "comprising" is used herein to describe and claim the present invention as synonymous with terms such as including, containing, or having, the invention, or embodiments thereof, may alternatively be described using alternative terms such as "consisting of" or "consisting essentially of."

[0067] Unless otherwise specified, the terms "a," "an," and "the" and similar referents when used in the context of describing particular embodiments of the present application (especially in the context of the claims) can be construed to cover both the singular and the plural. The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of individually referring to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated herein as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "etc."), provided with respect to certain embodiments herein is intended merely to better clarify the application and does not impose a limitation on the scope of the application as otherwise claimed. The abbreviation "eg," derived from the Latin "exempli gratia," is used herein to indicate a non-limiting example. Thus, the abbreviation "eg" is synonymous with the term "for example." No language in the specification should be construed as indicating any non-claimed element essential to the practice of the application.

[0068] "Optional" or "optionally" means that the subsequently described circumstance may or may not occur, so that the description includes instances in which the circumstance occurs and instances in which it does not occur.

[0069] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limiting. Members of each group may be referred to and claimed individually or in any combination with other members of the group or other elements found herein. One or more members of a group may be included in, or deleted from, a group for reasons of convenience and / or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified to satisfy all Markush group descriptions used in the appended claims.

[0070] The terms "treat," "treatment," "treating," or "amelioration," when used in reference to a disease, disorder, or medical condition, refer to both therapeutic and prophylactic or preventative measures, the purpose of which is to reverse, alleviate, improve, inhibit, alleviate, slow, or halt the progression or severity of a symptom or condition. The term "treating" includes reducing or alleviating at least one adverse effect or symptom of a condition. Treatment is generally "effective" if one or more symptoms or clinical markers are reduced. Alternatively, treatment is "effective" if the progression of a disease, disorder, or medical condition is reduced or halted. That is, "treatment" includes not only the improvement of symptoms or markers, but also the cessation of symptoms, or at least a slowing of their progression or worsening, that would be expected in the absence of treatment. "Treatment" can also mean pursuing or achieving a beneficial result or reducing an individual's likelihood of developing a condition, even if the treatment is ultimately unsuccessful. Those in need of treatment include those who already have the condition, as well as those prone to have the condition or those in whom the condition is to be prevented. Non-limiting examples of treatments or therapeutic treatments include at least one selected from pharmacological therapy, biological therapy, interventional surgical treatment, and combinations thereof. Non-limiting examples of therapeutic treatments include any one or more of coronary revascularization by stent insertion, coronary artery bypass grafting, or medical therapy, or combinations thereof. Non-limiting examples of medical therapy (blood pressure medications) include statins, LDL lowering, beta-blockers, angiotensin-converting enzyme (ACE) inhibitors, calcium channel blockers, angiotensin II antagonists, vasodilators, diuretics, renin inhibitors, aspirin, etc., or combinations thereof.

[0071] The term "prophylactic treatment" means maintaining or improving the healthy or disease-free state of a healthy or disease-free subject. The term "prophylactic treatment" also means preventing or slowing the appearance of symptoms associated with a condition, disease, or disorder. The term "prophylactic treatment" also means preventing or slowing a subject from acquiring a condition, disease, or disorder.

[0072] A "beneficial result" or "desired result" can include, but is not limited to, reducing or alleviating the severity of a condition, preventing a condition from worsening, curing a condition, preventing a condition from developing, reducing the likelihood that a patient will develop a condition, reducing morbidity and mortality, and increasing a patient's life expectancy or life expectancy. As non-limiting examples, a "beneficial result" or "desired result" can be the alleviation of one or more symptom(s), a reduction in the degree of deficit, a stabilized (i.e., not worsening) state of cardiovascular disease, a delay or slowing of cardiovascular disease, and an improvement or palliative relief of symptoms associated with cardiovascular disease.

[0073] The term "disease" refers to an abnormal condition affecting the body of an organism. The term "disorder" refers to a functional abnormality or disturbance. The terms disease and disorder are used interchangeably herein unless otherwise noted or clear given the context in which the terms are used.

[0074] As used herein, "disease," "pathological condition," and "condition" can include, but are in no way limited to, any form of cardiovascular condition, disease, or disorder. Cardiovascular disease is a type of disease that affects the heart or blood vessels. Non-limiting examples of cardiovascular disease include coronary artery disease, coronary heart disease, ischemic heart disease (IHD), cardiomyopathy, stroke, hypertensive heart disease, heart failure, pulmonary heart disease, ischemic syndrome, coronary microvascular disease, cardiac arrhythmia, rheumatic heart disease (RHD), aortic aneurysm, cardiomyopathy, atrial fibrillation, congenital heart disease, endocarditis, inflammatory heart disease, endocarditis, inflammatory cardiac hypertrophy, myocarditis, valvular heart disease, cerebrovascular disease, and peripheral arterial disease (PAD).

[0075] A "healthy subject" or "normal subject" is a subject that does not have a disease or disorder. In some embodiments, a healthy or normal subject does not have hypertension or a hypertensive emergency.

[0076] The term "administering" refers to placing an agent disclosed herein into a subject by a method or route that results in at least partial localization of the agent at the desired site. "Route of administration" can refer to any route of administration known in the art, including, but not limited to, aerosol, nasal, inhalation, oral, anal, intraanal, perianal, transmucosal, transdermal, parenteral, enteral, topical, or local. "Parenteral" refers to administration routes generally associated with injection, including intratumoral, intracranial, intraventricular, intrathecal, epidural, intradural, intraorbital, infusion, intravesical, intracardiac, intradermal, intramuscular, intraperitoneal, intrapulmonary, intraspinal, intrasternal, intrathecal, intrauterine, intravascular, intravenous, intraarterial, subarachnoid, subcapsular, subcutaneous, transmucosal, or transtracheal. Via the parenteral route, the composition can be in the form of a solution or suspension for infusion or injection, or as a lyophilized powder. Through the enteral route, the pharmaceutical composition can be in the form of a tablet, gel capsule, sugar-coated tablet, syrup, suspension, solution, powder, granule, emulsion, microsphere or nanosphere, or lipid vesicle or polymer vesicle, which allows controlled release.Through the topical route, the pharmaceutical composition can be in the form of an aerosol, lotion, cream, gel, ointment, suspension, solution, or emulsion.According to the present invention, "administering" can be self-administration.For example, a subject's consumption of the composition disclosed herein is considered to be "administering."

[0077] "Diagnosis" means identifying the presence or nature of a condition, disease, or disorder, including identifying patients at risk for developing a particular condition, disease, or disorder. Diagnostic methods vary in their sensitivity and specificity. The "sensitivity" of a diagnostic assay is the percentage of diseased individuals who test positive (percent of "true positives"). Diseased individuals not detected by the assay are "false negatives." Subjects who are not diseased and test negative in the assay are referred to as "true negatives." The "specificity" of a diagnostic assay is 1 minus the false positive rate, where the "false positive" rate is defined as the proportion of those without the disease who test positive. While a particular diagnostic method may not provide a definitive diagnosis of a condition, disease, or disorder, it suffices if the method provides a positive indication that aids in diagnosis.

[0078] "At risk of" is intended to mean that there is an increased risk compared to a normal subject or compared to a control group, e.g., a patient population. Thus, subjects carrying a particular marker may have an increased risk of a particular condition, disease, or disorder and may be identified as requiring further testing. "Increased risk" or "elevated risk" refers, for example, to any statistically significant increase in the probability that a subject will have a disorder. In some embodiments, the risk is increased by at least 10% relative to the control group being compared. In some embodiments, the risk is increased by at least 20% relative to the control group being compared. In some embodiments, the risk is increased by at least 50% relative to the control group being compared.

[0079] The terms "detection," "detecting," and the like may be used in the context of detecting a pathological condition, disease, or disorder (eg, when a positive assay result is obtained).

[0080] The term "diagnosis" or "dx" refers to the determination of the nature and cause of a particular phenomenon. Diagnosis typically refers to medical diagnosis, which is the process of determining a disease or disorder that explains the symptoms and signs. A diagnostic procedure, often a diagnostic test or assay, can be used to provide a diagnosis.

[0081] The term "prognosis" or "px" refers to predicting the likely outcome of a current condition. For example, a prognosis can include the expected duration and course of a disease or disorder, such as progressive decline or expected recovery.

[0082] The term "theranostics" or "tx" refers to diagnostics or prognostics used in the context of medical treatment. For example, theranostics can include diagnostic tests used to select appropriate and optimal therapies (or vice versa) based on the context of genetic material or other molecular or cellular analysis. Theranostics includes pharmacogenomics, personalized medicine, and precision medicine.

[0083] The term "sample" is used herein in its broadest sense. The term "biological sample" refers to a sample taken or isolated from an organism. Exemplary biological samples include, but are not limited to, buccal swabs, mucus, whole blood, blood, serum, plasma, urine, saliva, semen, lymph, fecal extracts, sputum, other bodily or biological fluids, cell samples, and tissue samples. The term also includes mixtures of the above samples. The term "sample" also includes unprocessed or preprocessed (or pre-processed) biological samples. In some embodiments, a sample may contain one or more cells from a subject. In some embodiments, the sample is a tissue or tissue sample. In some embodiments, the sample is a heart. In some embodiments, the sample is at least one portion of a heart. In some embodiments, the sample is at least one cardiac tissue. In some embodiments, the sample (e.g., heart) is examined non-invasively, and thus the sample is (within) a living subject.

[0084] "Subject" refers to a human or an animal. Typically, an animal is a vertebrate, such as a primate, a rodent, a domestic animal, or a game animal. Primates include chimpanzees, cynomolgus monkeys, spider monkeys, and macaques, e.g., rhesus monkeys. Rodents include mice, rats, woodchucks, ferrets, rabbits, and hamsters. Domestic and game animals include cattle, horses, pigs, deer, bison, water buffalo, feline species, e.g., domestic cats, and canine species, e.g., dogs, foxes, and wolves. The terms "patient," "individual," and "subject" are used interchangeably herein. In some embodiments, a subject is a mammal. A mammal may be a human, a non-human primate, a mouse, a rat, a dog, a cat, a horse, or a cow, but is not limited to these examples. Additionally, the methods described herein can be used to treat domestic animals and / or pets. In some embodiments, the subject is a human.

[0085] "Mammal" refers to any member of the class Mammalia, including, but not limited to, humans and non-human primates such as chimpanzees and other ape and monkey species; farm animals such as cows, sheep, pigs, goats, and horses; domestic mammals such as dogs and cats; and laboratory animals, including rodents such as mice, rats, and guinea pigs. The term does not denote a particular age or sex. Thus, adult and newborn subjects, as well as fetuses, whether male or female, are intended to be included within the scope of this term.

[0086] The subject may be a person who has been previously diagnosed with or identified as suffering from or having a condition requiring treatment (e.g., microhemorrhage, cardiovascular disease; or hypertension, hypertensive emergency) or one or more complications associated with that condition, and optionally, who has already received treatment for that condition or one or more complications associated with that condition. Alternatively, the subject may also be a person who has not previously been diagnosed with a condition or one or more complications associated with that condition. For example, the subject may be a person who exhibits one or more risk factors for a condition or one or more complications associated with that condition, or a subject who does not exhibit risk factors. A "subject in need" of treatment for a particular condition may be a subject suspected of having the condition, a subject diagnosed with the condition, a subject who has already been treated or is being treated for the condition, a subject who has not been treated for the condition, or a subject at risk of developing the condition.

[0087] "Microhemorrhage" refers to small bleeding likely caused by structural abnormalities in small blood vessels in an organ. In some embodiments, microhemorrhage refers to a condition in which red blood cells are found outside the vasculature, which may be due to abnormal permeability of the vasculature or rupture of blood vessels. Microhemorrhages typically occur adjacent to small blood vessels affected by amyloid angiopathy or hypertensive arteriopathy. In some cases, a subject may have persistent microhemorrhages, while in other cases, the microhemorrhages are acute microhemorrhages. In some embodiments, the methods disclosed herein are directed to the treatment, prevention, evaluation, and / or diagnosis of myocardial microhemorrhages, particularly in subjects with hypertension. In some embodiments, the methods disclosed herein are directed to the treatment, prevention, evaluation, and / or diagnosis of microhemorrhages, excluding cerebral microhemorrhages. In some embodiments, the methods disclosed herein are directed to myocardial or cardiac tissue, but not the brain. In some embodiments, the microhemorrhages in the methods disclosed herein are new or secondary microhemorrhages in myocardium or another tissue following an ischemic stroke.

[0088] Symptoms associated with microbleeds, particularly acute microbleeds, can include sudden tingling sensations (e.g., in one or both sides of the face, arms, chest, legs), heaviness in the arms and / or legs, double vision, and / or nausea.

[0089] "Hypertension," also known as high blood pressure, refers to blood pressure that is higher than normal (especially when measured repeatedly over time), where normal blood pressure levels are below 120 / 80 mmHg. In some embodiments, a subject with a systolic blood pressure of 130 mmHg or greater has hypertension. In some embodiments, a subject with hypertension also has a diastolic blood pressure of 80 mmHg or greater.

[0090] In various embodiments, the subject is an adult human and is diagnosed with hypertension if his or her blood pressure is 130 / 80 mmHg or higher. In some embodiments, a medical professional diagnoses a patient with hypertension if the patient's blood pressure is consistently 130 / 80 mmHg or higher, according to the American College of Cardiology / American Heart Association Guideline for the Prevention, Detection, Evaluation, and Management of High Blood Pressure in Adults (2017 guidelines). According to the 2017 guidelines, a patient is diagnosed with elevated blood pressure if they have a systolic blood pressure of 120-129 mmHg and a diastolic blood pressure of less than 80 mmHg. In some embodiments, a medical professional diagnoses a patient with hypertension if the patient's blood pressure is consistently 140 / 90 mmHg or higher, according to the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (2003 guidelines). According to the 2003 guidelines, patients are diagnosed as at risk or prehypertensive if they have a systolic blood pressure of 120-139 mmHg and a diastolic blood pressure of 80-89 mmHg.

[0091] "Hypertensive emergency" generally refers to an acute event that causes a transient increase in blood pressure due to psychological distress. An example of a hypertensive emergency is takotsubo cardiomyopathy (also known as stress cardiomyopathy).

[0092] In some embodiments, the subject is a child or adolescent human who is diagnosed with hypertension if his or her mean blood pressure, when measured multiple times over three or more clinic visits, is greater than or equal to the 95th percentile for the subject's age, sex, and height.

[0093] "Ferroptosis" refers to an iron-dependent form of cell death. The small molecule erastin inhibits cysteine ​​import, leading to glutathione depletion and inactivation of the phospholipid peroxidase glutathione peroxidase 4 (GPX4). GPX4 converts potentially harmful lipid hydroperoxides into non-toxic lipid alcohols. Inactivation of GPX4 by erastin or through GSH depletion by the direct GPX4 inhibitor (1S,3R)-RSL3 ultimately leads to overwhelming lipid peroxidation, which causes cell death. Iron is required for the accumulation of lipid peroxides and the execution of ferroptosis.Genes involved in the regulation of ferroptosis or serving as markers of ferroptosis include GXP4, acyl-CoA synthetase long-chain family member 4 (ACSL-4), aldo-keto reductase family 1 member C1 (AKR1C1-3), lipoxygenase, ATP synthase H+ transporting mitochondrial Fo complex subunit C3 (ATP5G3), cystathionine beta synthase (CBS), CD44 molecule, ChaC glutathione-specific gamma-glutamylcyclotransferase (GCTA). ferase 1 (CHAC1), CDGSH iron-sulfur domain 1 (CISD1), citrate synthase (CS), dipeptidyl-peptidase-4 (DPP4), Fanconi anemia complementation group D2 (FANCD2), glutamate-cysteine ​​ligase (GCLC / GCLM), glutaminase 2 (GLS2), glutathione synthetase (GSS), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), heat shock protein beta 1 / 5 (HSPB1 / 5), kiss of death (kiss These include kinase activator of death (KOD), lysophosphatidylcholine acyltransferase 3 (LPCAT3), metallothionein-1G (MT1G), nuclear receptor coactivator 4 (NCOA4), erythroid nuclear transcription factor 2-like 2 (NFE2L2), prostaglandin-endoperoxide synthase 2 (PTGS2), ribosomal protein L8 (RPL8), spermidine / spermine N1-acetyltransferase 1 (SAT2), solute carrier family 7 member 11 (SLC7A11), squalene synthase (SQS), transferrin receptor (TFRC), tumor protein 53 (TP53), and ER membrane protein complex subunit 2 (TT35 / EMC2). In some embodiments, the ferroptosis markers in the methods disclosed herein include any 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 of the genes involved in regulating ferroptosis, and can be measured using quantification of the or their DNA, mRNA, cDNA, or protein expression levels.

[0094] In various embodiments, the therapeutic agent may be provided as a pharmaceutical composition. In various embodiments, the pharmaceutical composition according to the present invention may be formulated for delivery via a route of administration. In certain embodiments, the pharmaceutical composition is formulated for intravascular, intravenous, or intra-arterial administration. In one embodiment, the pharmaceutical composition is formulated for intravenous administration as a single bolus.

[0095] In various embodiments, the pharmaceutical composition may contain any pharmaceutically acceptable excipient. "Pharmaceutically acceptable excipient" refers to an excipient that is generally safe, non-toxic, and useful in preparing a desirable pharmaceutical composition, including excipients acceptable for veterinary and human pharmaceutical use. Such excipients may be solid, liquid, semisolid, or, in the case of aerosol compositions, gaseous. Examples of excipients include, but are not limited to, starch, sugar, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrants, wetting agents, emulsifiers, colorants, release agents, coating agents, sweeteners, flavoring agents, fragrances, preservatives, antioxidants, plasticizers, gelling agents, thickeners, hardeners, setting agents, suspending agents, surfactants, humectants, carriers, stabilizers, and combinations thereof.

[0096] In various embodiments, the pharmaceutical composition according to the present invention may contain any pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle involved in carrying or transporting a compound of interest from one tissue, organ, or part of the body to another tissue, organ, or part of the body. For example, the carrier may be a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, or a combination thereof. Each component of the carrier must be "pharmaceutically acceptable" in that it must be compatible with the other components of the formulation. It must also be suitable for use in contact with any tissue or organ it may come into contact with; that is, it must not pose a risk of toxicity, irritation, allergic reaction, immunogenicity, or any other complication that unduly outweighs the benefits of its imaging.

[0097] The pharmaceutical compositions of the present invention can also be encapsulated, tableted, or prepared in emulsion or syrup for oral administration. Pharmaceutically acceptable solid or liquid carriers may be added to enhance or stabilize the composition or to facilitate preparation of the composition. Liquid carriers include syrup, peanut oil, olive oil, glycerin, saline, alcohol, and water. Solid carriers include starch, lactose, calcium sulfate, dihydrate, terra alba, magnesium stearate or stearic acid, talc, pectin, acacia, agar, or gelatin. The carrier may also include a sustained-release material, such as glyceryl monostearate or glyceryl distearate, alone or with a wax.

[0098] The pharmaceutical compositions are prepared according to conventional compounding techniques, such as dry milling, mixing, and compounding for powder forms; milling, mixing, granulating, and compressing (if necessary) for tablet forms; or milling, mixing, and filling for hard gelatin capsule forms. When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion, or aqueous or non-aqueous suspension. Such liquid preparations can be administered orally directly or by filling into soft gelatin capsules.

[0099] The pharmaceutical composition can be delivered in an amount effective to reduce or remove iron deposits from the heart, which can be accessed for efficacy via cardiac magnetic resonance imaging. The precise effective amount is the amount of composition that will produce the most effective results in terms of removing iron deposits and improving cardiac function and / or remodeling. This amount will vary depending on various factors, including, but not limited to, the characteristics of the therapeutic agent (including activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological state of the subject (age, sex, type and stage of disease, general health, responsiveness to a given dosage, and type of drug), the nature of the pharmaceutically acceptable carrier(s) in the formulation, and the route of administration. Those skilled in the clinical and pharmacological fields will be able to determine an amount effective for stress imaging through routine experimentation, for example, by monitoring image quality and adjusting dosage accordingly.

[0100] Before administration to a patient, a formulant may be added to the pharmaceutical composition. In some embodiments, the formulant is a liquid formulant. For example, these formulants may include oils, polymers, vitamins, carbohydrates, amino acids, salts, buffers, albumin, surfactants, bulking agents, or combinations thereof.

[0101] Carbohydrate formulating agents include sugars or sugar alcohols, such as monosaccharides, disaccharides, or polysaccharides, or water-soluble glucans. Sugars or glucans include fructose, dextrose, lactose, glucose, mannose, sorbose, xylose, maltose, sucrose, dextran, pullulan, dextrin, alpha- and beta-cyclodextrin, soluble starch, hydroxyethyl starch, and carboxymethylcellulose, or mixtures thereof. "Sugar alcohol" is defined as a C4-C8 hydrocarbon having an -OH group, including galactitol, inositol, mannitol, xylitol, sorbitol, glycerol, and arabitol. These sugars or sugar alcohols may be used individually or in combination. There is no set limit to the amount used, as long as the sugar or sugar alcohol is soluble in the aqueous preparation. In some embodiments, the sugar or sugar alcohol concentration is 1.0% to 7.0% w / v. In some embodiments, the sugar or sugar alcohol concentration is 2.0 to 6.0 w / v %.

[0102] Amino acid formulants include the levorotatory (L) forms of carnitine, arginine, and betaine, although other amino acids may be added.

[0103] Polymer formulants include polyvinylpyrrolidone (PVP) having an average molecular weight of 2,000 to 3,000, or polyethylene glycol (PEG) having an average molecular weight of 3,000 to 5,000.

[0104] A buffer may also be used in the pharmaceutical composition to minimize pH changes in the solution before lyophilization or after reconstitution. Almost any physiological buffer may be used, including, but not limited to, citrate, phosphate, succinate, and glutamate buffers, or mixtures thereof. In some embodiments, the concentration is 0.01 to 0.3 molar. Surfactants that can be added to the formulation are listed in European Patent Nos. 270,799 and 268,110.

[0105] Another drug delivery system for increasing circulation half-life is liposome.The preparation method of liposome delivery system is discussed in Gabizon et al., Cancer Research (1982) 42:4734, Cafiso, Biochem Biophys Acta (1981) 649:129, and Szoka, Ann Rev Biophys Eng (1980) 9:467.Other drug delivery systems are known in the art and are described, for example, in Poznansky et al., DRUG DELIVERY SYSTEMS (RL Juliano, ed., Oxford, NY 1980), pp.253-315, and ML Poznansky, Pharm Revs (1984) 36:277.

[0106] After the liquid pharmaceutical composition is prepared, it can be lyophilized to prevent degradation and preserve sterility. Methods for lyophilizing liquid compositions are known to those skilled in the art. Immediately before use, the composition can be reconstituted with a sterile diluent (e.g., Ringer's solution, distilled water, or sterile saline), which may contain additional ingredients. Once reconstituted, the composition is administered to a subject using methods known to those skilled in the art.

[0107] The pharmaceutical compositions of the present invention can be sterilized by conventional, well-known sterilization techniques. The resulting solution can be packaged for use or filtered and lyophilized under aseptic conditions, with the lyophilized preparation being combined with a sterile solution prior to administration. The compositions may contain pharmaceutically acceptable auxiliary substances, such as pH adjusting and buffering agents, tonicity adjusting agents, etc., such as sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, and stabilizers (e.g., 1-20% maltose), as needed to approximate physiological conditions.

[0108] Many variations and alternative elements are disclosed in the embodiments of the present invention. Still further variations and alternative elements will be apparent to those skilled in the art. Among these variations are, but are not limited to, the steps for the methods, pharmaceutical compositions, routes of administration and devices, the selection of imaging techniques, and the diseases and other clinical conditions that can be diagnosed, prognosed, or treated therewith. Various embodiments of the present invention can specifically include or exclude any of these variations or elements.

[0109] In some embodiments, numbers expressing properties such as amounts and concentrations of ingredients, reaction conditions, and the like, used to describe and claim certain embodiments of the present invention are to be understood as being modified in some instances by the term "about." Accordingly, in some embodiments, the numerical parameters set forth in the specification and appended claims are approximations that may vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the present invention are approximations, the numerical values ​​set forth in the specific examples are reported as precisely as practicable. The numerical values ​​presented in some embodiments of the present invention may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements.

[0110] Many people suffer from or are at risk of developing hypertension. Hypertension can lead to a variety of adverse outcomes, including cardiovascular disease, cardiac arrhythmias, sudden cardiac death, and death. It is not known how hypertension leads to heart failure or other major adverse cardiovascular events.

[0111] Applicants have discovered that microbleeds, detected as iron deposits, occur in the hearts of subjects with hypertension. Specifically, Applicants have discovered that microbleeds, detected as iron deposits, occur in the myocardium of spontaneously hypertensive rats. This is an important finding because the presence of iron in the myocardium may be a major mediator of adverse cardiac remodeling and fatal outcomes in patients with hypertension or hypertensive emergencies. Applicants' discovery that microbleeds occur in hypertensive hearts and lead to iron deposition in the myocardium may help provide an understanding of the pathophysiology of cardiac remodeling in hypertension.

[0112] Accordingly, it is a non-limiting object of the present invention to provide methods for detecting, diagnosing, and / or treating microhemorrhages in the heart so as to reduce adverse outcomes in people with hypertension.

[0113] Various Non-Limiting Embodiments of the Invention In various embodiments, the method includes the heart of the subject. In some embodiments, the heart is a human heart. In some embodiments, the subject is a human.

[0114] In various embodiments, the method includes administering at least one portion of the subject's heart.

[0115] In various embodiments, the method includes administering at least one cardiac tissue of a subject. In some embodiments, the cardiac tissue is human cardiac tissue. In some embodiments, the subject is a human.

[0116] In various embodiments, the method includes a heart of a control subject. In some embodiments, the heart is a human heart. In some embodiments, the control subject is a human.

[0117] In various embodiments, the method includes measuring at least one portion of a heart of a control subject. In some embodiments, the heart is a human heart. In some embodiments, the control subject is a human.

[0118] In various embodiments, the method includes at least one cardiac tissue of a control subject. In some embodiments, the cardiac tissue is human cardiac tissue. In some embodiments, the control subject is a human.

[0119] In various embodiments, methods are provided for identifying and / or assessing at least one microhemorrhage in a subject's heart, the methods including obtaining at least one clinical image (e.g., SPECT, PET, CT, ultrasound, or magnetic resonance image) of the subject's heart and detecting at least one iron deposit in the subject's heart from the at least one clinical image, wherein the detection of the iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart. In various embodiments, the subject has or is diagnosed with hypertension. In some embodiments, the detection of at least one iron deposit in the subject's heart identifies the subject as having myocardial microhemorrhage. In further embodiments, the detection of at least one iron deposit in the subject's heart indicates the subject has hypertension. In further embodiments, a subject identified as having myocardial microhemorrhage has a higher content of adipocyte infiltration compared to a reference sample, particularly if the subject has had myocardial microhemorrhage or hypertension for at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 1 year, 2 years, 3 years, 5 years, or longer.

[0120] In various embodiments, a method is provided for detecting at least one microhemorrhage in a subject's heart, the method comprising obtaining at least one clinical image (e.g., SPECT, PET, CT, ultrasound, or magnetic resonance image) of the subject's heart and detecting at least one iron deposit in the subject's heart from the at least one clinical image, wherein the detection of the at least one iron deposit in the subject's heart is indicative of and / or is the detection of at least one microhemorrhage in the subject's heart.

[0121] In various embodiments, a method for prognosing at least one microhemorrhage in a subject's heart is provided, the method comprising obtaining at least one clinical image of the subject's heart and detecting at least one iron deposit in the subject's heart from the at least one clinical image, wherein the detection of the at least one iron deposit in the subject's heart is prognostic of at least one microhemorrhage in the subject's heart.

[0122] In various embodiments, a method is provided for diagnosing at least one microhemorrhage in a subject's heart, the method comprising obtaining at least one clinical image of the subject's heart and detecting at least one iron deposit in the subject's heart from the at least one clinical image, wherein the detection of the at least one iron deposit in the subject's heart is diagnostic of at least one microhemorrhage in the subject's heart.

[0123] In various embodiments, a method is provided for assessing and / or determining a subject's risk of developing at least one microhemorrhage in the heart, the method comprising obtaining at least one clinical image of the subject's heart and detecting at least one iron deposit in the subject's heart from the at least one image, wherein detection of the at least one iron deposit in the subject's heart indicates an increased risk of developing at least one microhemorrhage in the subject's heart.

[0124] In various embodiments, a method is provided for identifying and / or assessing at least one microhemorrhage in at least one portion of a subject's heart, the method comprising obtaining at least one clinical image of the at least one portion of the subject's heart and detecting at least one iron deposit in the at least one portion of the subject's heart from the at least one clinical image, wherein detection of the at least one iron deposit in the at least one portion of the subject's heart is indicative of at least one microhemorrhage in the at least one portion of the subject's heart.

[0125] In various embodiments, a method is provided for detecting at least one microhemorrhage in at least one portion of a subject's heart, the method comprising obtaining at least one clinical image of at least one portion of the subject's heart and detecting at least one iron deposit in the at least one portion of the subject's heart from the at least one clinical image, wherein the detection of the at least one iron deposit in the at least one portion of the subject's heart is indicative of and / or is the detection of at least one microhemorrhage in the at least one portion of the subject's heart.

[0126] In various embodiments, a method for prognosing at least one microhemorrhage in at least one portion of a subject's heart is provided, the method comprising obtaining at least one clinical image of at least one portion of the subject's heart and detecting at least one iron deposit in the at least one portion of the subject's heart from the at least one clinical image, wherein the detection of the at least one iron deposit in the at least one portion of the subject's heart is prognostic of at least one microhemorrhage in the at least one portion of the subject's heart.

[0127] In various embodiments, a method is provided for diagnosing at least one microhemorrhage in at least one portion of a subject's heart, the method comprising obtaining at least one clinical image of at least one portion of the subject's heart and detecting at least one iron deposit in the at least one portion of the subject's heart from the at least one clinical image, wherein the detection of the at least one iron deposit in the at least one portion of the subject's heart is diagnostic of at least one microhemorrhage in the at least one portion of the subject's heart.

[0128] In various embodiments, a method is provided for assessing and / or determining a subject's risk of developing at least one microhemorrhage in at least one portion of the subject's heart, the method comprising obtaining at least one clinical image of at least one portion of the subject's heart and detecting at least one iron deposit in the at least one portion of the subject's heart from the at least one clinical image, wherein detection of the at least one iron deposit in the at least one portion of the subject's heart indicates an increased risk of the subject for developing at least one microhemorrhage in the at least one portion of the subject's heart.

[0129] In various embodiments, a method is provided for identifying and / or assessing at least one microhemorrhage in at least one cardiac tissue of a subject, the method comprising obtaining at least one clinical image of the at least one cardiac tissue and detecting at least one iron deposit in the at least one cardiac tissue from the at least one image, wherein detection of the at least one iron deposit in the at least one cardiac tissue is indicative of at least one microhemorrhage in the at least one cardiac tissue of the subject.

[0130] In various embodiments, a method is provided for detecting at least one microhemorrhage in at least one cardiac tissue of a subject, the method comprising obtaining at least one clinical image of the at least one cardiac tissue and detecting at least one iron deposit in the at least one cardiac tissue from the at least one clinical image, wherein the detection of the at least one iron deposit in the at least one cardiac tissue is indicative of and / or is the detection of at least one microhemorrhage in the at least one cardiac tissue of the subject.

[0131] In various embodiments, a method is provided for prognosing at least one microhemorrhage in at least one cardiac tissue of a subject, the method comprising obtaining at least one clinical image of the heart and detecting at least one iron deposit in the heart from the at least one clinical image, wherein the detection of the at least one iron deposit in the heart is prognostic of at least one microhemorrhage in the subject's heart.

[0132] In various embodiments, a method is provided for diagnosing at least one microhemorrhage in at least one cardiac tissue of a subject, the method comprising obtaining at least one clinical image of the at least one cardiac tissue and detecting at least one iron deposit in the at least one cardiac tissue from the at least one clinical image, wherein the detection of the at least one iron deposit in the at least one cardiac tissue is diagnostic of at least one microhemorrhage in the at least one cardiac tissue of the subject.

[0133] In various embodiments, a method is provided for assessing and / or determining a subject's risk of developing at least one microhemorrhage in at least one cardiac tissue, the method comprising obtaining at least one clinical image of the at least one cardiac tissue and detecting at least one iron deposit in the at least one cardiac tissue from the at least one clinical image, wherein detection of the at least one iron deposit in the at least one cardiac tissue indicates an increased risk of the subject for developing at least one microhemorrhage in the subject's at least one cardiac tissue.

[0134] In various embodiments, a method is provided for identifying and / or assessing at least one microhemorrhage in a subject's heart, the method comprising obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying the presence of at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart; and comparing the clinical image of the subject's heart with at least one clinical image from a reference sample, wherein the presence of the at least one iron deposit in the subject's heart compared to the reference sample is indicative of at least one microhemorrhage in the subject's heart.

[0135] In various embodiments, a method for detecting at least one microhemorrhage in a subject's heart is provided, the method comprising the steps of obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying the presence of at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart; and comparing the clinical image of the subject's heart with at least one clinical image from a reference sample, wherein the presence of the at least one iron deposit in the subject's heart compared to the reference sample is indicative of at least one microhemorrhage in the subject's heart.

[0136] In various embodiments, a method for prognosing at least one microhemorrhage in a subject's heart is provided, the method comprising obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart; and comparing the clinical image of the subject's heart with at least one clinical image from a reference sample, wherein the presence of the at least one iron deposit in the subject's heart compared to the reference sample is prognostic of the at least one microhemorrhage in the subject's heart.

[0137] In various embodiments, a method is provided for diagnosing at least one microhemorrhage in a subject's heart, the method comprising obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart; and comparing the clinical image of the subject's heart with at least one clinical image from a reference sample, wherein the presence of the at least one iron deposit in the subject's heart compared to the reference sample is diagnostic of at least one microhemorrhage in the subject's heart.

[0138] In various embodiments, a method is provided for assessing and / or determining a subject's risk of developing at least one microhemorrhage in the heart, the method comprising the steps of obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying at least one iron deposit in the heart from the at least one clinical image of the subject's heart; and comparing the clinical image of the subject's heart with at least one clinical image from a reference sample, wherein the presence of the at least one iron deposit in the subject's heart compared to the reference sample indicates an increased risk of the subject for developing at least one microhemorrhage in the subject's heart.

[0139] In various embodiments, a method is provided for determining progression of at least one microhemorrhage in a subject's heart, the method comprising the steps of obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart; and comparing the clinical image of the subject's heart with at least one clinical image from a reference sample, wherein a change in the clinical image of the subject's heart compared to the at least one clinical image from the reference sample is indicative of progression of at least one microhemorrhage in the subject's heart.

[0140] In various embodiments, a method for identifying and / or assessing cardiovascular disease in a subject is provided, the method comprising: obtaining at least one clinical image of the subject's heart; detecting at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart, wherein detection of the iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the clinical image of the subject's heart with at least one clinical image from a reference sample, wherein a change in the clinical image of the subject's heart compared to the at least one clinical image from the reference sample is indicative of cardiovascular disease in the subject.

[0141] In various embodiments, a method for detecting cardiovascular disease in a subject is provided, the method comprising: obtaining at least one clinical image of the subject's heart; detecting at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart, wherein detection of the at least one iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the at least one clinical image of the subject's heart with at least one clinical image from a reference sample, wherein a change in the at least one clinical image of the subject's heart compared to the at least one clinical image from the reference sample is indicative of cardiovascular disease in the subject.

[0142] In various embodiments, a method for detecting cardiovascular disease in a subject is provided, the method comprising: obtaining at least one clinical image of the subject's heart; detecting at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart, wherein detection of the at least one iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the at least one clinical image of the subject's heart with at least one clinical image from a reference sample, wherein a change in the at least one clinical image of the subject's heart compared to the at least one clinical image from the reference sample is indicative of cardiovascular disease in the subject.

[0143] In various embodiments, a method for prognosing cardiovascular disease in a subject is provided, the method comprising: obtaining at least one clinical image of the subject's heart; detecting at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart, wherein the detection of iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the clinical image of the subject's heart with at least one clinical image from a reference sample, wherein the change in the clinical image of the subject's heart compared with the at least one clinical image from the reference sample is a prognostic prediction of cardiovascular disease in the subject.In some embodiments, the subject with myocardial microhemorrhage has a worse cardiac remodeling outcome (for example, in terms of worse cardiac function) or a longer remodeling period compared with the control subject.

[0144] In various embodiments, a method for diagnosing cardiovascular disease in a subject is provided, the method comprising: obtaining at least one clinical image of the subject's heart; detecting at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart, wherein detection of the iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the clinical image of the subject's heart with at least one clinical image from a reference sample, wherein a change in the clinical image of the subject's heart compared to the at least one clinical image from the reference sample is diagnostic of cardiovascular disease in the subject.

[0145] In various embodiments, a method for assessing and / or determining a risk of developing cardiovascular disease in a subject is provided, the method comprising the steps of obtaining at least one clinical image of the subject's heart; detecting at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart, wherein the detection of the iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the clinical image of the subject's heart with at least one clinical image from a reference sample, wherein a change in the clinical image of the subject's heart compared to the at least one clinical image from the reference sample indicates an increased risk of the subject for developing cardiovascular disease in the subject.

[0146] In various embodiments, a method for determining the progression of cardiovascular disease in a subject is provided, the method comprising the steps of obtaining at least one clinical image of the subject's heart; detecting at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart, wherein the detection of the iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's heart; and comparing the clinical image of the subject's heart with at least one clinical image from a reference sample, wherein a change in the clinical image of the subject's heart compared to the at least one clinical image from the reference sample indicates the progression of cardiovascular disease in the subject.

[0147] In various embodiments, a method is provided for identifying at least one biomarker for cardiovascular disease in a subject, the method comprising: obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying at least one iron deposit in the subject's heart from the at least one clinical image of the subject's heart; correlating the at least one iron deposit in the at least one clinical image of the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject with a biomarker signature from a reference sample, wherein at least one biomarker for cardiovascular disease is identified.

[0148] In various embodiments, a method for identifying and / or assessing cardiovascular disease in a subject is provided, the method comprising obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying at least one iron deposit in the subject's heart from the at least one clinical image; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject to a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is indicative of cardiovascular disease in the subject.

[0149] In various embodiments, a method for detecting cardiovascular disease in a subject is provided, the method comprising obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying at least one iron deposit in the subject's heart from the at least one clinical image; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject with a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is indicative of and / or detection of cardiovascular disease in the subject.

[0150] In various embodiments, a method for prognosing cardiovascular disease in a subject is provided, the method comprising obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying at least one iron deposit in the subject's heart from the at least one clinical image; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject to a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is prognostic of cardiovascular disease in the subject.

[0151] In various embodiments, a method for diagnosing cardiovascular disease in a subject is provided, the method comprising obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying at least one iron deposit in the subject's heart from the at least one clinical image; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject to a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is diagnostic of cardiovascular disease in the subject.

[0152] In various embodiments, a method is provided for determining the progression of cardiovascular disease in a subject, the method comprising obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying at least one iron deposit in the subject's heart from the at least one clinical image; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject with a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is indicative of the progression of cardiovascular disease in the subject.

[0153] In some embodiments, there is provided a method for assessing and / or determining a risk of developing cardiovascular disease in a subject, the method comprising: obtaining at least one clinical image of the subject's heart; detecting and / or measuring and / or quantifying at least one iron deposit in the subject's heart from the at least one clinical image; correlating the at least one iron deposit in the subject's heart with at least one microhemorrhage in the subject's heart to obtain a biomarker signature for the subject; and comparing the biomarker signature from the subject with a biomarker signature from a reference sample, wherein a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is indicative of an increased risk of the subject for developing cardiovascular disease.

[0154] In various embodiments, a method is provided for assessing and / or determining the risk of at least one cardiac arrhythmia in a subject, the method comprising obtaining at least one clinical image of the subject's heart and detecting the presence of at least one iron deposit in the subject's heart from the at least one clinical image, wherein the presence of the at least one iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and wherein the at least one microhemorrhage in the subject's heart is indicative of an increased risk of at least one cardiac arrhythmia in the subject.

[0155] In various embodiments, a method is provided for assessing and / or determining risk of sudden cardiac death in a subject, the method comprising obtaining at least one clinical image of the subject's heart and detecting the presence of at least one iron deposit in the subject's heart from the at least one clinical image, wherein the presence of the at least one iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and wherein the at least one microhemorrhage in the subject's heart is indicative of an increased risk of sudden cardiac death in the subject.

[0156] In various embodiments, a method is provided for assessing and / or determining risk of cardiovascular disease in a subject, the method comprising obtaining at least one clinical image of the subject's heart and detecting the presence of at least one iron deposit in the subject's heart from the at least one clinical image, wherein the presence of the at least one iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and wherein the at least one microhemorrhage in the subject's heart is indicative of an increased risk of cardiovascular disease in the subject.

[0157] Various embodiments provide methods for identifying, detecting, prognosing, and / or diagnosing at least one microhemorrhage in a subject's heart, the method comprising detecting and / or measuring the presence of at least one iron deposit in the subject's heart by performing a cardiac single-photon emission computed tomography (SPECT) scan, wherein the presence of the iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and the subject has been diagnosed with hypertension, hypertensive emergency, acute hypertensive emergency, persistent hypertension, or a combination thereof. In some embodiments, the method comprises identifying myocardial microhemorrhages in a subject, the method comprises performing a SPECT scan of the subject's heart or myocardium, respectively, to detect the presence of iron deposits in the heart or myocardium, the presence of which is indicative of at least one myocardial microhemorrhage. In some embodiments, the method comprises detecting myocardial microhemorrhages in a subject, the method comprises performing a SPECT scan of the subject's heart or myocardium, respectively, to detect the presence of iron deposits in the heart or myocardium, the presence of which is indicative of at least one myocardial microhemorrhage. In some embodiments, a method for prognosing myocardial microhemorrhage in a subject comprises performing a SPECT scan of the subject's heart or myocardium to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which indicates at least one myocardial microhemorrhage.In some embodiments, a method for diagnosing myocardial microhemorrhage in a subject comprises performing a SPECT scan of the subject's heart or myocardium to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which indicates at least one myocardial microhemorrhage.

[0158] In some embodiments, SPECT scan is replaced with or performed in addition to measuring the level of hemoglobin degradation product or ferroptosis marker in the blood of the subject, and the presence of this product or marker indicates at least one myocardial microhemorrhage.In some embodiments, this is for the method of identifying microhemorrhage in the heart or myocardium of the subject.In some embodiments, this is for the method of detecting microhemorrhage in the heart or myocardium of the subject.In some embodiments, this is for the method of prognosticating microhemorrhage in the heart or myocardium of the subject.In some embodiments, this is for the method of diagnosing microhemorrhage in the heart or myocardium of the subject.

[0159] Various embodiments provide methods for identifying, detecting, prognosing, and / or diagnosing at least one microhemorrhage in the myocardium of a subject, the method comprising detecting and / or measuring the presence of at least one iron deposit in the subject's heart by performing a single photon emission computed tomography (SPECT) scan of the subject's myocardium, wherein the presence of the iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's myocardium or heart, and the subject has been diagnosed with hypertension, hypertensive emergency, acute hypertensive emergency, persistent hypertension, or a combination thereof.

[0160] Various embodiments provide methods for identifying, detecting, prognosing, and / or diagnosing at least one microhemorrhage in a subject's heart, the method comprising detecting and / or measuring the presence of at least one iron deposit in the subject's heart by performing a cardiac positron emission tomography (PET) scan, wherein the presence of the iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and the subject has been diagnosed with hypertension, hypertensive emergency, acute hypertensive emergency, persistent hypertension, or a combination thereof. In some embodiments, the method comprises identifying cardiac or myocardial microhemorrhages in a subject, the method comprises performing a PET scan of the subject's heart or myocardium, respectively, to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage. In some embodiments, the method comprises detecting cardiac or myocardial microhemorrhages in a subject, the method comprises performing a PET scan of the subject's heart or myocardium, respectively, to detect the presence of iron deposits in the heart or myocardium, the presence of which is indicative of at least one myocardial microhemorrhage. In some embodiments, there is provided a method for prognosing cardiac or myocardial microhemorrhage in a subject, comprising performing a PET scan of the subject's heart or myocardium to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage. In some embodiments, there is provided a method for diagnosing cardiac or myocardial microhemorrhage in a subject, comprising performing a PET scan of the subject's heart or myocardium to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage.

[0161] Various embodiments provide methods for identifying, detecting, prognosing, and / or diagnosing at least one microhemorrhage in a subject's heart, the method comprising detecting and / or measuring the presence of at least one iron deposit in the subject's heart by performing a cardiac computed tomography (CT) scan, wherein the presence of the iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and the subject has been diagnosed with hypertension, hypertensive emergency, acute hypertensive emergency, persistent hypertension, or a combination thereof. In some embodiments, the method comprises identifying cardiac or myocardial microhemorrhages in a subject, the method comprises performing a CT scan of the subject's heart or myocardium, respectively, to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage. In some embodiments, the method comprises detecting cardiac or myocardial microhemorrhages in a subject, the method comprises performing a CT scan of the subject's heart or myocardium, respectively, to detect the presence of iron deposits in the heart or myocardium, the presence of which is indicative of at least one myocardial microhemorrhage. In some embodiments, there is provided a method for prognosing cardiac or myocardial microhemorrhage in a subject, comprising performing a CT scan of the subject's heart or myocardium to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage. In some embodiments, there is provided a method for diagnosing cardiac or myocardial microhemorrhage in a subject, comprising performing a CT scan of the subject's heart or myocardium to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage.

[0162] Various embodiments provide methods for identifying, detecting, prognosing, and / or diagnosing at least one microhemorrhage in a subject's heart, the method comprising detecting and / or measuring the presence of at least one iron deposit in the subject's heart by performing a cardiac ultrasound (US) scan, wherein the presence of the iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and the subject has been diagnosed with hypertension, hypertensive emergency, acute hypertensive emergency, persistent hypertension, or a combination thereof. In some embodiments, the method comprises identifying cardiac or myocardial microhemorrhages in a subject, the method comprises performing a US scan of the subject's heart or myocardium, respectively, to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage. In some embodiments, the method comprises detecting cardiac or myocardial microhemorrhages in a subject, the method comprises performing a US scan of the subject's heart or myocardium, respectively, to detect the presence of iron deposits in the heart or myocardium, the presence of which is indicative of at least one myocardial microhemorrhage. In some embodiments, there is provided a method of prognosing cardiac or myocardial microhemorrhage in a subject, comprising performing a US scan of the subject's heart or myocardium to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage. In some embodiments, there is provided a method of diagnosing cardiac or myocardial microhemorrhage in a subject, comprising performing a US scan of the subject's heart or myocardium to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage.

[0163] Various embodiments provide methods for identifying, detecting, prognosing, and / or diagnosing at least one microhemorrhage in a subject's heart, the method comprising detecting and / or measuring the presence of at least one iron deposit in the subject's heart by performing a cardiac magnetic resonance imaging (MRI) scan, wherein the presence of the iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and the subject has been diagnosed with hypertension, hypertensive emergency, acute hypertensive emergency, persistent hypertension, or a combination thereof. In some embodiments, the method comprises identifying cardiac or myocardial microhemorrhages in a subject, the method comprises performing an MRI scan of the subject's heart or myocardium, respectively, to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage. In some embodiments, the method comprises detecting cardiac or myocardial microhemorrhages in a subject, the method comprises performing an MRI scan of the subject's heart or myocardium, respectively, to detect the presence of iron deposits in the heart or myocardium, the presence of which is indicative of at least one myocardial microhemorrhage. In some embodiments, there is provided a method for prognosing cardiac or myocardial microhemorrhage in a subject, comprising performing an MRI scan of the subject's heart or myocardium to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage. In some embodiments, there is provided a method for diagnosing cardiac or myocardial microhemorrhage in a subject, comprising performing an MRI scan of the subject's heart or myocardium to detect the presence of iron deposits in the heart or myocardium, respectively, the presence of which is indicative of at least one cardiac or myocardial microhemorrhage.

[0164] Various embodiments provide a method for identifying, detecting, prognosing, and / or diagnosing at least one microhemorrhage in a subject's heart, the method comprising detecting and / or measuring the presence of at least one iron deposit in the subject's heart and measuring a level of a hemoglobin breakdown product from the subject's blood, wherein the presence of the iron deposit in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and the subject has been diagnosed with hypertension, a hypertensive emergency, an acute hypertensive emergency, persistent hypertension, or a combination thereof. Exemplary hemoglobin breakdown products include iron, bilirubin, globin, and combinations thereof.

[0165] Various embodiments provide methods for identifying, detecting, prognosing, and / or diagnosing at least one microhemorrhage in a subject's heart, the methods comprising: detecting and / or measuring the presence of at least one iron deposit in the subject's heart; and measuring a level of a ferroptosis marker from the subject's blood, wherein the presence of iron deposits in the subject's heart is indicative of at least one microhemorrhage in the subject's heart, and the subject has been diagnosed with hypertension, hypertensive emergency, acute hypertensive emergency, persistent hypertension, or a combination thereof. Exemplary ferroptosis markers include those listed in connection with the description of ferroptosis above. In some embodiments, the ferroptosis marker in the methods comprises glutathione peroxidase 4 (GPX4). A method for identifying cardiac or myocardial microhemorrhage in a subject is provided, the method comprising or consisting of measuring a level of a ferroptosis marker, including GPX4, from the subject's blood, wherein the presence of GPX4 or a higher level of GPX4 than a reference subject is indicative of cardiac or myocardial microhemorrhage in the subject. A method for detecting cardiac or myocardial microhemorrhage in a subject is provided, the method comprising or consisting of measuring the level of a ferroptosis marker, including GPX4, from the subject's blood, wherein the presence of GPX4 or a higher level of GPX4 than a reference subject indicates cardiac or myocardial microhemorrhage in the subject.A method for prognosing cardiac or myocardial microhemorrhage in a subject is provided, the method comprising or consisting of measuring the level of a ferroptosis marker, including GPX4, from the subject's blood, wherein the presence of GPX4 or a higher level of GPX4 than a reference subject indicates cardiac or myocardial microhemorrhage in the subject.A method for diagnosing cardiac or myocardial microhemorrhage in a subject is provided, the method comprising or consisting of measuring the level of a ferroptosis marker, including GPX4, from the subject's blood, wherein the presence of GPX4 or a higher level of GPX4 than a reference subject indicates cardiac or myocardial microhemorrhage in the subject.In a further embodiment, the ferroptosis markers in the method further comprise another 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 markers in addition to GPX4.

[0166] In various embodiments, particularly for cardiac tissue, the technique of detecting iron using MRI is T2 * MRI sequences selected from T1, T2, proton density weighted sequences, and susceptibility weighted imaging (SWI) are used, thereby obtaining T1 maps, T1 weighted images, susceptibility weighted images including quantitative susceptibility maps, T2 maps, and T2 * Map, T2 or T2 * Map, T2 or T2 * Weighted, corrected T2 or T2 * The method includes obtaining an enhanced image. Other techniques for detecting iron in cardiac tissue include histological staining of cardiac tissue specimens or several invasive techniques, such as inductively coupled plasma mass spectrometry. Details of detecting iron or iron deposits in cardiac tissue are described in U.S. Pat. No. 10,694,961, which is incorporated herein by reference in its entirety.

[0167] In some embodiments, the subject is a human. In some embodiments, the subject has at least one selected from the group consisting of hypertension, a hypertensive emergency, an acute hypertensive emergency, persistent hypertension, and combinations thereof.

[0168] In some embodiments, the subject has not had a myocardial infarction or reperfusion therapy-induced hemorrhage, and in some further aspects, the subject has not had a hemorrhagic myocardial infarction, and in some further aspects, the subject has not undergone reperfusion therapy and therefore does not have reperfusion hemorrhage. In some embodiments, the subject has not had a myocardial infarction or undergone reperfusion therapy in the past 3 days, 7 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 2 years, 3 years, or 5 years. In some embodiments, a subject in need of one or more of the methods disclosed herein has any one or more of the above characteristics, and in some embodiments, a subject in need thereof desires identification, detection, prognosis, or diagnosis of myocardial microhemorrhage.

[0169] In some embodiments, the subject has idiopathic hypertension. In some embodiments, the subject has long-term or chronic idiopathic hypertension that develops hypertensive microhemorrhages. In some embodiments, the subject has symptoms, e.g., transient focal symptoms associated with acute microhemorrhages, such as sudden tingling sensations (e.g., in one or both sides of the face, arms, chest, or limbs), heaviness in the arms and / or legs, double vision, and / or nausea. In some embodiments, the subject has been diagnosed with hypertension. In some embodiments, the subject has been diagnosed with hypertension and is taking an anticoagulant (e.g., apixaban, dabigatran, edoxaban, enoxaparin, heparin, rivaroxaban, warfarin). In some embodiments, the subject has been diagnosed with a stress-induced condition (cardiomyopathy), such as takotsubo cardiomyopathy.

[0170] In some embodiments, methods for treating a subject with myocardial microhemorrhage are provided. In some embodiments, one or more of the methods disclosed above for identifying, detecting, prognosing, or diagnosing myocardial microhemorrhage, or thereby associated cardiovascular disease, further include treating the subject with a therapeutic agent and / or selecting a therapeutic agent for the subject and / or providing a therapeutic agent to the subject and / or administering a therapeutic agent to the subject. In some embodiments, the therapeutic agent is a therapeutic composition comprising at least one chelating agent. In some embodiments, the chelating agent is selected from the group consisting of deferoxamine (also known as desferrioxamine), deferasirox, deferiprone, hinokitiol, dexrazoxane, 2,2'-bipyridyl, pyridoxal isonicotinoyl hydrazone, salicylaldehyde isonicotinoyl hydrazone, exochelin (including desferri-exochelin), and combinations thereof. Desferriexochelins are hexadentate molecules that form a one-to-one binding relationship with iron, thereby preventing free radical reactions, while deferiprone is a bidentate molecule and deferasirox is a tridentate molecule. In some embodiments, the chelator is an iron chelator. In some embodiments, the iron chelator is selected from the group consisting of an intracellular iron chelator, an extracellular iron chelator, or a combination thereof. In some embodiments, the intracellular iron chelator chelates ferrous iron, ferric iron, and a combination thereof. In some embodiments, the extracellular iron chelator chelates ferrous iron, ferric iron, and a combination thereof. In some embodiments, the therapeutic agent is selected from the group consisting of at least one heme-binding protein, at least one heme-degrading protein, and a combination thereof. In some embodiments, the method further comprises administering to the subject an amount of an agent effective to increase the amount of at least one selected from the group consisting of at least one heme-binding protein, at least one heme-degrading protein, at least one heme-blocking protein, and a combination thereof in the subject.In some embodiments, the heme-binding protein is selected from the group consisting of hemopexin, haptoglobin, albumin, ferritin, alpha1-microglobulin, alpha1-antitrypsin, glutathione-S-transferase, heme-binding protein / liver fatty acid-binding protein, heme-binding protein 23 / peroxiredoxin, p22 heme-binding protein, and glyceraldehyde-3-phosphate dehydrogenase, and combinations thereof. In some embodiments, the heme-degrading protein is selected from the group consisting of heme oxygenase 1, hinokitiol, nuclear factor E2-related factor 2 (Nrf2), and combinations thereof. In some embodiments, a heme-blocking protein refers to a protein that binds to a protein that otherwise transports heme, reducing the uptake of heme into cells and thereby neutralizing the function of the protein to which it is bound, thus indirectly blocking heme. For example, the heme-blocking agent may be an inhibitor of heme carrier protein 1 (HCP1) or an inhibitor of divalent metal transporter 1 (DMT1). In another example, the heme blocker is hepcidin, which binds to ferroportin, and the complex undergoes endocytosis and degradation, resulting in the loss of ferroportin from the cell surface and thus inducing a secondary decrease in iron release from the cell. In some embodiments, the factor is selected from FLVCR1a (feline leukemia virus subgroup C receptor 1a), ABCG2 (ATP-binding cassette subfamily G member 2), FLVCR2, and combinations thereof. In some embodiments, the heme-binding protein is a heme scavenger.

[0171] Additional agents can be included or administered in combination with the therapeutic composition, including, but not limited to, anti-inflammatory agents, lipid-lowering agents, agents that can promote iron clearance by enhancing macrophage activity, phagocytosis-enhancing agents, or agents that can interfere with the biosynthesis of iron oxide crystals or prevent nanocrystal aggregation, or combinations thereof. When administered "in combination," the agents can be premixed before administration, administered simultaneously, or administered sequentially (including consecutively or separated by administration-free periods). Representative anti-inflammatory agents include colchicine, corticosteroids, nonsteroidal anti-inflammatory drugs (NSAIDs), anti-IL-1 beta (e.g., anakinra), anti-TNF-α (e.g., etanercept and infliximab), anti-IL-6 (e.g., tocilizumab), anti-MMP (e.g., PG-116800 and doxycycline), macrophage modulators (e.g., phosphatidylserine-presenting liposomes), NLRP3 inflammasome inhibitors (e.g., 16673-34-0 (5-chloro-2-methoxy-N-[2-(4-sulfamoylphenyl)ethyl]benzamide)), inflammasome antagonists (e.g., P2X7 antagonists), or antidiabetic agents (e.g., insulin (e.g., Humulin, Novolin, Humalog), metformin (e.g., Glycerin), These include steroids such as steroids (e.g., steroids for steroid use, e ...Representative lipid-lowering agents include statins, cholesterol absorption inhibitors (eg, ezetimbie), bile acid binding resins / sequestrants (eg, cholestyramine), niacin, or vitamin B3.

[0172] In some embodiments, the chelator, heme-binding protein, heme-blocking protein, and / or heme-degrading protein is administered for a period of 1 day to 1 year, e.g., 1 week, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 1 year, or for at least 1 year, 2 years, 3 years, 4 years, or 5 years.

[0173] In some embodiments, the therapeutic composition or treatment is initiated or administered to the subject immediately after the onset of symptoms or within 5 minutes, 10 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 12 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 1 month of the onset of symptoms. In further embodiments, the therapeutic composition or treatment is initiated or administered to the subject after the subject receives a diagnosis confirming the presence of myocardial iron deposits or microhemorrhages.

[0174] Further embodiments provide that the treatment method also includes monitoring or reassessing the subject. In some embodiments, the method includes imaging the subject's heart after administration of the composition (one dose) to detect levels of iron deposits in the myocardium or other parts of the heart. In further embodiments, the method includes administering a subsequent dose of the composition to the subject if the myocardium or other parts of the heart exhibits the same or higher levels of iron deposits compared to the levels before the subject was administered the previous dose, or discontinuing administration of the composition if no iron deposits are present in the myocardium or other parts of the heart. In further embodiments, myocardial iron levels of 1-5 μg / g tissue, 5-10 μg / g tissue, 0.01 mg / g tissue, 0.02 mg / g tissue, 0.03 mg / g tissue, 0.04 mg / g tissue, or greater indicate that the subject has developed or recurring myocardial microhemorrhage.

[0175] In the above methods, exemplary dosages of the chelating agent, heme-binding protein, heme-degrading protein, heme-blocking protein, or factor disclosed above per unit body weight of the subject include 10-100 μg, 100-200 μg, 200-300 μg, 300-400 μg, 400-500 μg, 500-600 μg, 600-700 μg, 700-800 μg, 800-900 μg, 900-1000 μg, 1000-1200 μg, 1300-1400 μg, 1400-1500 μg, 1500-1600 μg, 1600-1700 μg, 1700-1800 μg, 1800-2000 μg, 1900-2100 μg, 2000-2200 μg, 2100-2400 μg, 2200-2400 μg, 2300-2500 μg, 2400-2600 μg, 2500-3000 μg, 2600-3200 μg, 2700-3400 μg, 2800-3600 μg, 2900-3800 μg, 3000-3900 μg, 3100-3200 μg, 3300-3400 μg, 3500-3600 μg, 3600-3800 μg, 3700-3800 μg, 3800-4000 μg, 3900-4100 μg, 3900-4200 μg, The dosages include 00-900 μg, 1-5 mg, 5-10 mg, 10-20 mg, 20-30 mg, 30-40 mg, 40-50 mg, 50-60 mg, 60-70 mg, 70-80 mg, 80-90 mg, 90-100 mg, 100-200 mg, 200-300 mg, 300-400 mg, 400 mg-500 mg, 500 mg-1 g, or 1 g-10 g. The unit weight of the subject can be per kg of body weight or per subject. In another embodiment, the chelating agent (e.g., deferoxamine, deferasirox, deferiprone) may be used at any one or more of the following doses: 100-200 mg / period, 200-300 mg / period, 300-400 mg / period, 400-500 mg / period, 500-600 mg / period, 600-700 mg / period, 700-800 mg / period, 800-900 mg / period, 900-100 mg / period, or combinations thereof. The dosing period may be daily, within 24 hours, 48 ​​hours, 96 hours, weekly, every two weeks, or monthly.

[0176] In some embodiments, the iron deposits comprise iron oxide. In some embodiments, the iron deposits comprise at least one compound comprising iron. In some embodiments, the compound is selected from the group consisting of heme, heme derivatives, and combinations thereof. In some embodiments, the iron is in any possible oxidation state. In some embodiments, the oxidation state is selected from +2, +2 / +3, +3, +4, and combinations thereof. In some embodiments, the iron is selected from the group consisting of ferrous iron, ferric iron, and combinations thereof. In some embodiments, the iron is in any form.

[0177] In some embodiments, the portion of the heart is selected from the group consisting of epicardium, myocardium, endocardium, valve tissue, and combinations thereof. In some embodiments, the portion of the heart is selected from the group consisting of valve tissue, endocardium, pulmonary valve, brachiocephalic trunk, superior vena cava, right pulmonary artery, right pulmonary vein, tricuspid valve, right ventricle, inferior vena cava, descending aorta, papillary muscle, interventricular septum, epicardium, left ventricle, myocardium, mitral valve, aortic valve, left pulmonary vein, left pulmonary artery, aorta, left subclavian artery, pulmonary trunk, left common carotid artery, and combinations thereof.

[0178] In some embodiments, the change is in at least one iron deposit, hi some embodiments, the change is in at least one selected from the group consisting of amount, quantity, concentration, shape, size, and pattern of the at least one iron deposit.

[0179] In some embodiments, the reference sample is the heart, heart tissue or part of the heart from a control subject, and the control subject does not have at least one iron deposit in the heart, heart tissue or part of the heart.In some embodiments, the reference sample is the heart, heart tissue or part of the heart from a control subject, and the control subject does not have at least one microhemorrhage in the heart, heart tissue or part of the heart.In some embodiments, the reference sample is the heart, heart tissue or part of the heart from a subject at an earlier time point.

[0180] In some embodiments, the cardiovascular disease is selected from the group consisting of infarcted myocardium, coronary artery disease, coronary heart disease, ischemic heart disease, cardiomyopathy, stroke, hypertensive heart disease, heart failure, pulmonary heart disease, ischemic syndrome, coronary microvascular disease, cardiac arrhythmia, rheumatic heart disease, aortic aneurysm, cardiomyopathy, atrial fibrillation, congenital heart disease, endocarditis, inflammatory heart disease, endocarditis, inflammatory cardiac hypertrophy, myocarditis, valvular heart disease, cerebrovascular disease, peripheral arterial disease, acute stress-induced cardiomyopathy (takotsubo cardiomyopathy), diffuse myocardial fibrosis, and combinations thereof.

[0181] In some embodiments, the reference sample is a heart, heart tissue, or a portion of a heart from a subject before the subject is treated for cardiovascular disease, hi some embodiments, the reference sample is a heart, heart tissue, or a portion of a heart from a subject who has been successfully treated for cardiovascular disease.

[0182] In some embodiments, the reference sample is a heart, heart tissue, or a portion of a heart from a control subject, wherein the control subject does not have cardiovascular disease. In some embodiments, the reference sample is a heart, heart tissue, or a portion of a heart from a control subject, wherein the control subject has cardiovascular disease.

[0183] In some embodiments, the reference sample is from the heart, heart tissue, or portion of the heart of a subject who has been successfully treated for cardiovascular disease, hi some embodiments, the reference sample is from the heart, heart tissue, or portion of the heart of a subject at an earlier time point.

[0184] In some embodiments, obtaining at least one magnetic resonance image comprises operating a magnetic resonance imaging machine / scanner to obtain the at least one magnetic resonance image.

[0185] In some embodiments, the cardiac tissue is selected from the group consisting of valve tissue, endocardium, pulmonary valve, brachiocephalic artery, superior vena cava, right pulmonary artery, right pulmonary vein, tricuspid valve, right ventricle, inferior vena cava, descending aorta, papillary muscle, interventricular septum, epicardium, left ventricle, myocardium, mitral valve, aortic valve, left pulmonary vein, left pulmonary artery, aorta, left subclavian artery, pulmonary trunk, left common carotid artery, and combinations thereof. [Example]

[0186] The present invention is further illustrated by the following examples, which are intended to be purely exemplary of the present invention and should not be construed as limiting the present invention in any way. The following examples are provided to better illustrate the claimed invention and should not be construed as limiting the scope of the present invention. To the extent that specific materials are mentioned, they are for illustrative purposes only and are not intended to limit the present invention. Those skilled in the art may develop equivalent means or reactants without the exercise of inventive capacity and without departing from the scope of the present invention.

[0187] Example 1 FIG. 1A shows iron in myocardial tissue in wild-type rats treated with angiotensin II to induce hypertension, and FIG. 1B shows the resulting myocardial fibrosis.

[0188] To provide aspects of the present disclosure, embodiments may employ any number of programmable processing devices executing software or stored instructions. Physical processors and / or machines used by embodiments of the present disclosure for any processing or evaluation may include one or more networked (Internet, cloud, WAN, LAN, satellite, wired, or wireless (RF, cellular, WiFi, Bluetooth, etc.)) or non-networked general-purpose computer systems, microprocessors, field programmable gate arrays (FPGAs), digital signal processors (DSPs), microcontrollers, smart devices (e.g., smartphones), computer tablets, handheld computers, etc., programmed according to the teachings of the exemplary embodiments. Additionally, the devices and subsystems of the exemplary embodiments may be implemented by the preparation of application-specific integrated circuits (ASICs) or by interconnecting an appropriate network of conventional component circuits. Thus, the exemplary embodiments are not limited to any specific combination of hardware circuitry and / or software.

[0189] When stored on any one or combination of computer-readable media, exemplary embodiments of the present disclosure may include software for controlling the devices and subsystems of the exemplary embodiments, for driving the devices and subsystems of the exemplary embodiments, for enabling the devices and subsystems of the exemplary embodiments to interact with a human user, etc. Such software may include, but is not limited to, device drivers, firmware, operating systems, development tools, application software, database management software, etc. The computer code devices of the exemplary embodiments may include any suitable interpretable or executable code mechanism, including, but not limited to, scripts, interpretable programs, dynamic link libraries (DLLs), Java classes and applets, full executable programs, etc. Moreover, processing power may be distributed across multiple processors for better performance, reliability, cost, or other benefits.

[0190] Common forms of computer-readable media may include, for example, floppy disks, flexible disks, hard disks, magnetic tape, any other suitable magnetic media, CD-ROMs, CDRWs, DVDs, any other suitable optical media, punch cards, paper tape, optical mark sheets, any other suitable physical media having a pattern of holes or other optically recognizable indicia, RAM, PROMs, EPROMs, FLASH-EPROMs, any other suitable memory chips or cartridges, carrier waves, or any other suitable media from which a computer can read. Such storage media may also be used to store other types of data, such as data organized in databases, for access, processing, and communication by a processing device.

[0191] The various methods and techniques described above provide several means for implementing the present application. Of course, it should be understood that not necessarily all of the described objects or advantages can be achieved in accordance with any particular embodiment described herein. Thus, for example, one skilled in the art will recognize that the methods can be implemented in a manner that achieves or optimizes one advantage or group of advantages taught herein without necessarily achieving other objects or advantages as taught or suggested herein. Various alternatives are mentioned herein. It should be understood that some embodiments specifically include one, another, or several features, while other embodiments specifically exclude one, another, or several features, and still other embodiments reduce a particular feature by including one, another, or several advantageous features.

[0192] Additionally, those skilled in the art will recognize the applicability of various features from different embodiments. Similarly, the various elements, features, and steps discussed above, as well as other known equivalents to each such element, feature, or step, may be used in various combinations by those skilled in the art to implement methods in accordance with the principles described herein. Some of the various elements, features, and steps are specifically included, while others are specifically excluded, in various embodiments.

[0193] Although the present application has been disclosed in terms of certain embodiments and examples, it will be understood by those skilled in the art that the embodiments of the present application extend beyond the specifically disclosed embodiments to other alternative embodiments and / or uses, and modifications and equivalents thereof.

[0194] Various embodiments of the present application are described herein, including the best mode known to the inventors for carrying out the application. Variations on these embodiments will become apparent to those skilled in the art upon reading the foregoing description. Those skilled in the art will be able to employ such variations as appropriate, and it is contemplated that the present application may be practiced otherwise than as specifically described herein. Accordingly, many embodiments of the present application include all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by this application unless otherwise indicated herein or clearly contradicted by context.

[0195] All patents, patent applications, published patent applications, and other materials, such as articles, books, specifications, publications, documents, objects, and / or the like, referenced herein are hereby incorporated by reference in their entirety for all purposes, except for any intermediate processing file history associated therewith, either of which is inconsistent or contradictory with this specification, or which may have a limiting effect on the broadest scope of any claims now or hereafter associated with this specification. By way of example, in the event of a conflict or inconsistency between the explanation, definition, and / or use of a term associated with any of the incorporated materials and that associated with this specification, the explanation, definition, and / or use of the term in this specification shall control.

[0196] It should be understood that the embodiments of the present application disclosed herein are illustrative of the principles of the embodiments of the present application. Other modifications that may be employed may be within the scope of the present application. Thus, by way of example, and not of limitation, alternative configurations of the embodiments of the present application may be utilized in accordance with the teachings herein. Accordingly, the embodiments of the present application are not limited to the exact ones shown and described.

[0197] Various embodiments of the present invention have been described above in the detailed description. While these descriptions directly describe the above embodiments, it should be understood that those skilled in the art may envision modifications and / or variations to the specific embodiments shown and described herein. Any such modifications or variations that fall within the scope of this specification are also intended to be included herein. Unless otherwise noted, it is the inventors' intention that the words and phrases in this specification and claims be given the ordinary and accustomed meaning to those of ordinary skill in the applicable technical field(s).

[0198] The foregoing description of various embodiments of the invention known to applicant at the time of filing this application has been presented and is intended for purposes of illustration and description. It is not intended to be exhaustive or to limit the invention to the precise form disclosed, many modifications and variations being possible in light of the above teachings. The described embodiments serve to illustrate the principles of the invention and its practical application, and to enable those skilled in the art to utilize the invention in various embodiments and with various modifications as suited to the particular use contemplated. Therefore, it is intended that the invention not be limited to the particular embodiments disclosed for carrying out the invention.

[0199] While particular embodiments of the present invention have been shown and described, it will be apparent to those skilled in the art that, based on the teachings herein, changes and modifications may be made without departing from this invention and its broader aspects, and it is therefore intended that the appended claims encompass within their scope all such changes and modifications that are within the true spirit and scope of this invention.

Claims

1. 1. A method for identifying and / or detecting at least one microhemorrhage in the myocardium of a subject, comprising: detecting and / or measuring the presence of at least one iron deposit in the heart of the subject by performing cardiac imaging, wherein the subject has been diagnosed with hypertension, a hypertensive emergency, an acute hypertensive emergency, persistent hypertension, or a combination thereof. Including, the presence of said iron deposits in the heart of said subject is indicative of at least one microhemorrhage in the myocardium of said subject. The method.

2. 10. The method of claim 1, wherein the subject has not had a myocardial infarction or reperfusion therapy-induced hemorrhage, or the subject has not had a myocardial infarction or reperfusion therapy-induced hemorrhage in the past 3 days, 7 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 2 years, 3 years, or 5 years.

3. 3. The method of claim 1 or 2, wherein the detecting and / or measuring comprises performing imaging of the subject's heart, the imaging being selected from the group consisting of single photon emission computed tomography (SPECT), positron emission tomography (PET), computed tomography (CT), ultrasound (US), magnetic resonance imaging (MRI), and combinations thereof.

4. The method of claim 3 , wherein the detecting and / or measuring comprises performing a SPECT scan, a PET scan, or both, of the subject's heart.

5. 5. The method of claim 1, wherein the step of detecting and / or measuring the presence of at least one iron deposit further comprises measuring a level of a hemoglobin degradation product or a level of a ferroptosis marker from the subject's blood, wherein the hemoglobin degradation product is selected from the group consisting of iron, bilirubin, globin, and combinations thereof, and the ferroptosis marker comprises glutathione peroxidase 4 (GPX4).

6. comparing images of the subject's heart with those obtained from a reference sample; The method of any one of claims 1 to 5, further comprising:

7. 7. The method of claim 6, wherein the presence of the at least one iron deposit in the subject's heart compared to the reference sample indicates at least one microhemorrhage in the subject's myocardium.

8. a change in the image of the subject's heart compared to that obtained from the reference sample is indicative of the progression of at least one microhemorrhage in the subject's myocardium.

7. The method of claim 6 for detecting the progression of at least one microhemorrhage in the myocardium of the subject.

9. 1. A method for identifying and / or detecting cardiovascular disease or the progression of said cardiovascular disease in a subject, comprising: detecting at least one iron deposit in the subject's heart by performing cardiac imaging, wherein the subject has been diagnosed with hypertension, hypertensive emergency, acute hypertensive emergency, persistent hypertension, or a combination thereof, and wherein the detection of the iron deposit in the subject's heart indicates the presence of at least one microhemorrhage in the subject's myocardium; comparing an image of the subject's heart with one obtained from a reference sample, wherein a change in the image of the subject's heart compared to that obtained from the reference sample is indicative of cardiovascular disease in the subject or is indicative of the progression of the cardiovascular disease; The method comprising:

10. correlating the at least one iron deposit in the at least one image of the subject's heart with at least one microhemorrhage in the subject's myocardium to obtain a biomarker signature from the at least one image of the subject. further comprising the comparing step compares the biomarker signature obtained from the image of the subject with a biomarker signature from a reference sample, thereby identifying at least one biomarker for cardiovascular disease; a change in the biomarker signature from the subject compared to the biomarker signature from the reference sample is indicative of the cardiovascular disease in the subject, or is the detection of the cardiovascular disease, or is indicative of the progression of the cardiovascular disease.

10. The method of claim 9.

11. 11. The method of claim 9 or 10, wherein the subject has not had a myocardial infarction or reperfusion therapy-induced hemorrhage, or the subject has not had a myocardial infarction or reperfusion therapy-induced hemorrhage in the past 3 days, 7 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 2 years, 3 years, or 5 years.

12. The method of any one of claims 9 to 11, wherein the cardiovascular disease comprises cardiomyopathy, myocardial fibrosis, cardiac arrhythmia, or sudden cardiac death.

13. A pharmaceutical composition for treating a subject experiencing, recovering from, and / or at risk of developing cardiovascular disease associated with myocardial microbleeds, comprising: the pharmaceutical composition comprises a chelating agent, a heme-binding protein, a heme-degrading protein, a heme-blocking agent, an agent effective to increase the amount of a heme-binding protein or a heme-degrading protein, or a combination thereof; the subject has hypertension, a hypertensive emergency, an acute hypertensive emergency, chronic hypertension, or a combination thereof; the presence of at least one iron deposit in the heart of the subject is detected and / or measured, whereby cardiovascular disease is associated with microhemorrhages in the myocardium; the detecting and / or measuring includes performing imaging of the heart. The pharmaceutical composition.

14. the chelating agent is selected from the group consisting of deferoxamine, deferasirox, deferiprone, hinokitiol, dexrazoxane, 2,2'-bipyridyl, and combinations thereof; the heme-binding protein is selected from the group consisting of hemopexin, haptoglobin, albumin, ferritin, alpha 1-microglobulin, alpha 1-antitrypsin, glutathione-S-transferase, heme-binding protein / liver fatty acid-binding protein, heme-binding protein 23 / peroxiredoxin, p22 heme-binding protein, and glyceraldehyde-3-phosphate dehydrogenase, and combinations thereof; the heme-degrading protein is selected from the group consisting of heme oxygenase 1, nuclear factor E2-related factor 2 (Nrf2), and combinations thereof; the heme blocker comprises hepcidin, and / or The factor is selected from feline leukemia virus subgroup C receptor (FLVCR) 1a (FLVCR1a), FLVCR2, ATP-binding cassette subfamily G member 2 (ABCG2), and combinations thereof.

14. The pharmaceutical composition of claim 13.

15. 14. The pharmaceutical composition of claim 13, wherein the subject's heart is imaged after administration of the composition to detect levels of iron deposition in the myocardium or other parts of the heart.

16. a subsequent dose of the composition is administered to the subject if the subject exhibits the same or higher level of iron deposition in the myocardium or other portion of the heart compared to the level before the previous dose was administered to the subject, or administration of the composition is discontinued if there are no iron deposits in the myocardium or other portion of the heart.

16. The pharmaceutical composition of claim 15,

17. The pharmaceutical composition according to any one of claims 13 to 15, wherein the subject is a human.

18. 18. The pharmaceutical composition of any one of claims 13 to 17, wherein the subject has not had a myocardial infarction or reperfusion therapy-induced hemorrhage, or the subject has not had a myocardial infarction or reperfusion therapy-induced hemorrhage in the past 3 days, 7 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 2 years, 3 years, or 5 years.

19. The method of any one of claims 1 to 8, for identifying at least one microhemorrhage in the myocardium of the subject.

20. The method of any one of claims 1 to 8, for detecting at least one microhemorrhage in the myocardium of the subject.

21. The method of any one of claims 9 to 12, for identifying said cardiovascular disease in said subject or for identifying a subject having said cardiovascular disease.

22. The method according to any one of claims 9 to 12, for detecting said cardiovascular disease in said subject.

23. The method according to any one of claims 9 to 12, for detecting the progression of said cardiovascular disease in said subject.

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