[go: up one dir, main page]

WO2011029201A1 - Méthodes et kits pour la détection de cardiopathie - Google Patents

Méthodes et kits pour la détection de cardiopathie Download PDF

Info

Publication number
WO2011029201A1
WO2011029201A1 PCT/CA2010/001426 CA2010001426W WO2011029201A1 WO 2011029201 A1 WO2011029201 A1 WO 2011029201A1 CA 2010001426 W CA2010001426 W CA 2010001426W WO 2011029201 A1 WO2011029201 A1 WO 2011029201A1
Authority
WO
WIPO (PCT)
Prior art keywords
level
sample
subject
mindin
hacel
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CA2010/001426
Other languages
English (en)
Inventor
Sara Arab
Peter Liu
Mark Suchul Moon
Liyong Zhang
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University Health Network
University of Toronto
Original Assignee
University Health Network
University of Toronto
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University Health Network, University of Toronto filed Critical University Health Network
Publication of WO2011029201A1 publication Critical patent/WO2011029201A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/32—Cardiovascular disorders
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/32—Cardiovascular disorders
    • G01N2800/324—Coronary artery diseases, e.g. angina pectoris, myocardial infarction

Definitions

  • the disclosure relates to methods and compositions for the detection of heart disease and specifically to the use of biomarkers and compositions comprising agents that bind the biomarkers for the detection of heart disease.
  • Mindin (Spondin 2) is a highly conserved secreted extracellular matrix (ECM) protein of the Mindin-F-spondin family. Previous studies showed Mindin to be a regulator of host innate immunity, but despite its high expression in the heart, its role in cardiac stress response is unknown.
  • ECM extracellular matrix
  • Hace is a HECT family E3 ubiquitin-protein ligase gene, HACE1 , and a potential suppressor of multiple human cancers. Genetic inactivation of HACE1 in mice results in the development of spontaneous, late onset tumors (Zhang, 2007).
  • Biomarkers useful for diagnosing heart disease are disclosed herein.
  • Treatments targeting the biomarkers are useful for treating heart disease.
  • mice that have a loss of function for Mindin show lower mortality and morbidity post myocardial infarction.
  • Minden expression is associated with higher mortality and acute adverse remodelling post myocardial infarction.
  • Minden levels are increased in human serum of ischemic subjects compared to controls.
  • the disclosure provides a method of screening for, diagnosing or detecting heart disease in a subject, the method comprising: a. determining a level of Mindin in a sample of the subject; and b. comparing the level of Mindin in the sample with a control; wherein an increased level of Mindin in the sample compared to the control is indicative that the subject has heart disease.
  • the method comprises: a) obtaining a sample from the subject;
  • the disclosure provides a method for treating a subject having heart disease comprising:
  • a determining a level of Mindin in a sample from the subject, and b. treating the subject with a treatment for heart disease when the level of Mindin in the sample is increased compared to control.
  • Yet a further aspect provides a method of monitoring response to treatment comprising: a. determining a base-line level of Mindin in a base-line sample from the subject;
  • Another aspect provides a method of monitoring heart disease progression comprising: a. determining a base-line level of Mindin in a base-line sample from the subject;
  • an increase in the Mindin level in the post-base-line sample compared to the base-line level is indicative the heart disease is progressing, and a decrease in the biomarker level in the post base-line sample compared to the baseline level is indicative that the heart disease is not progressing.
  • the heart disease is ischemia. In another embodiment the heart disease dilated cardiomyopathy.
  • the level of Mindin is determined using a biomarker specific reagent, such an antibody or antibody fragment, which specifically binds Mindin.
  • Hacel is useful for differentiating systolic heart failure from diastolic heart failure.
  • Hacel gene and protein expression levels are significantly elevated in blood samples of systolic heart failure patients. Further Hacel knock out mice fail to develop hypertrophy following cardiac banding.
  • the disclosure provides a method of screening for, diagnosing or detecting heart disease in a subject, the method comprising: a. determining a level of Hacel in a sample from the subject; and b. comparing the level of Hacel in the sample with a control; wherein an increased level of Hacel in the sample compared to the control is indicative that the subject has heart disease.
  • the disclosure provides a method for treating a subject having heart disease comprising:
  • c. determining a level of Hacel in a sample from the subject, and d. treating the subject with a treatment for heart disease when the level of Hacel in the sample is increased compared to control.
  • Yet a further aspect provides a method of monitoring response to treatment comprising:
  • Another aspect provides a method of monitoring heart disease progression comprising: a. determining a base-line level of Hacel in a base-line sample from the subject; b. determining a level of Hacel in a sample taken subsequent to the base-line sample from the subject; and
  • an increase in the Hacel level in the post-base-line sample compared to the base-line level is indicative the heart disease is progressing, and a decrease in the biomarker level in the post base-line sample compared to the baseline level is indicative that the heart disease is not progressing.
  • the heart disease is systolic heart failure.
  • the disclosure provides a method of distinguishing systolic heart failure from diastolic heart failure in a subject with heart failure, the method comprising:
  • a determining a level of Hacel in a sample from the subject, and b. comparing the level of Hacel in the sample with a control, wherein the control is a level of Hacel in a subject without heart failure or a subject with diastolic heart failure;
  • an increased level of Hacel in the sample compared to the control is indicative the subject has systolic heart failure and wherein a level comparable in the sample to the control is indicative the subject has diastolic heart failure.
  • the level of Hacel is determined using a biomarker specific reagent, such an antibody or antibody fragment, which specifically binds Hacel .
  • the sample and/or control comprises polypeptide.
  • the sample and/or control is a blood sample and comprises blood or a fraction thereof such as serum.
  • the treatment is an agent that inhibits the biomarker such as an antibody specific for the biomarker or a siRNA molecule that inhibits the expression of the biomarker, for example injected in heart or administered intravenously.
  • an agent that inhibits the biomarker such as an antibody specific for the biomarker or a siRNA molecule that inhibits the expression of the biomarker, for example injected in heart or administered intravenously.
  • a further aspect provides a kit for detecting Mindin, and/or Hacel levels.
  • FIG. 3 Western blots shows HACE1 protein levels is elevated in blood samples of systolic heart failure patient but not in blood samples from diastolic heart failure patient. DHF-diastolic heart failure; SHF-systolic heart failure.
  • FIG. 1 Human heart tissue microarray showing Mindin imRNA expression.
  • FIG. 1 Western blott and graph showing Mindin expression ICM and DCM human serum.
  • screening for, diagnosing or detecting ischemia refers to a method or process of determining if a subject has or does not have heart disease.
  • subject refers to any member of the animal kingdom, preferably a human being including for example a subject that has or is suspected of having heart disease such as heart failure.
  • the term "level” as used herein refers to an amount or quantity (e.g. relative amount or concentration) of biomarker that is detectable or measurable in a sample.
  • the level can be a concentration such as pg/L or a relative amount such as 1.1 , 1.2, 1.3, 1.4, 1 .5, 1.6, 1.7, 1 .8, 1.9, 2.0, 2.2, 2.4, 2.6, 2.8, 3.0, 3.2, 3.4, 3.6, 3.8, 4.0, 4.2, 4.4, 4.6, 4.8, 5.0, 10, 15, 20, 25, 30, 40, 60, 80 and/or 100 times a control level, where for example, the control level is the level such as the average or median level in a normal sample (e.g.
  • the level is the level of polypeptide. In another embodiment, the level is the level of mRNA or cDNA.
  • the term "determining a level" as used in reference to a biomarker means the application of a method to a sample, for example a sample of the subject and/or a control sample, for ascertaining quantitatively, semi-quantitatively or qualitatively the amount of a biomarker, for example the amount of biomarker polypeptide or mRNA.
  • a level of a biomarker can be determined by a number of methods including for example mass spectrometric methods, including for example MS, MS/MS, LC- MS/MS, SRM etc where a peptide of a biomarker is labeled and the amount of labeled biomarker peptide is ascertained, immunoassays including for example immunohistochemistry, Western blotting, ELISA, immunoprecipitation and the like, where a biomarker biomarker specific reagent such as an antibody for example, a labeled antibody specifically binds the biomarker and permits for example relative or absolute ascertaining of the amount of polypeptide biomarker, hybridization and PCR protocols, where a probe or primer or primer set are used to ascertain the amount of nucleic acid biomarker, including for example RT-PCR.
  • mass spectrometric methods including for example MS, MS/MS, LC- MS/MS, SRM etc where a peptide of a biomarker is labeled and the amount of
  • the term "increase in the level” as used herein refers to an increase in the level or quantity of a biomarker described herein, for example Mindin or Hacel , in a sample from the subject that is measurable compared to a suitable control and/or reference.
  • the term “decrease in the level” as used herein refers to a decrease in the level or quantity of a biomarker described herein, for example Mindin or Hacel , in a sample from the subject that is measurable compared to a suitable control and/or reference
  • the increase or decrease can be an increase or decrease in the steady-state level of a gene transcript, including for example a difference resulting from a difference in the level of transcription and/or translation and/or transcript degradation.
  • the increase or decrease in the level can refer to an increase or decrease in the measurable polypeptide or fragment thereof level of a given biomarker as measured by the amount of steady state level of and/or expressed polypeptide or fragment thereof in a test sample as compared with the measurable expression level of a given biomarker or fragment thereof in a control, population of control samples and/or previously taken or reference sample, for example in a blood sample.
  • the difference in the level can refer to an increase or decrease in the measurable nucleic acid transcript level of a given biomarker as measured by the amount of transcript e.g. biomarker mRNA or cDNA, for example as detectable in a sample comprising heart tissue.
  • an increase or decrease in the level can refer to an increase or decrease in the level of the biomarker in the subsequent sample (e.g. post treatment sample) compared to a reference sample (e.g. baseline sample), wherein an increase is indicative of negative therapeutic response and/or disease progression and a decrease is indicative of a positive therapeutic response and/or disease stabilization or amelioration.
  • a level of biomarker level is detected if a ratio of the level in a test sample as compared with a control is greater than or less than 1 .0 and/or if the ratio of the level in a reference sample as compared with a subsequent sample is greater than or less than 1 .0.
  • control refers to a sample from an subject or a group of individuals who are known as not having heart disease or to a biomarker level or value reflective of individuals who are known as not having heart disease, at which or below which individuals are likely to not have heart disease or the particular subtype being determined by the method (e.g. diastolic heart failure).
  • control is optionally derived from tissue of the same type as the sample of the subject being tested.
  • the control can be a serum sample where the sample from the subject being tested is a serum sample.
  • the numerical value or range is a predetermined value or range that corresponds to a level of the biomarker or range of levels of the biomarker in a group of subjects known as not having heart disease (e.g. threshold or cutoff level; or control range) or a particular subtype of heart disease (e.g. ischemic heart disease).
  • the control can be a cut-off or threshold level, above or below which (depending on the biomarker and subtype) which a subject is identified as having heart disease or a particular type thereof. For example, a test subject that has an increased level of a biomarker above a cut-off or threshold level is indicated to have or is more likely to have heart disease or a particular type of heart disease.
  • the term "positive control” as used herein refers to a sample of an individual or a group of individuals with heart disease and/or a value e.g. corresponding to a level of one or more biomarkers associated with the disease class, e.g. systolic heart failure, cardiac ischemia, post-myocardial infarction etc.
  • the term "baseline level” as used herein refers to a level that is used for comparison to a sample taken at a later time point. For example, in methods related to monitoring response to treatment or disease progression, “base-line level” can refer to a level of a biomarker in a sample taken prior to a subsequent sample, e.g. base line sample is taken before treatment, comparison to which provides an indication of response to treatment.
  • biomarker includes without limitation, a polypeptide or fragment, a polypeptide or fragment thereof that is secreted or released, for example which is secreted or released into blood (e.g. soluble biomarker), a nucleic acid sequence including a gene, or corresponding RNA, for example a RNA level in a heart tissue sample.
  • the biomarkers of the disclosure include for example, Mindin, Hacel , their homologs and fragments thereof.
  • polypeptide biomarker refers to polypeptide and/or fragments thereof of a biomarker of the present disclosure and includes polypeptides translated from the RNA transcripts of biomarkers described herein. Polypeptide biomarkers include modified (e.g. post-translational modifications such as glycosylation), expressed, as well as soluble biomarkers such as secreted, cleaved, released, and shed polypeptide products.
  • polypeptide and “protein” are intended to be used interchangeably.
  • soluble biomarker refers to a polypeptide biomarker, that is released in any manner from a cell and detectable in a blood sample, such as blood, serum, plasma.
  • prognosis refers to an expected clinical outcome group such as a poor survival group or a good survival group associated with or reflected by an increased biomarker level or levels when compared to a control, wherein the biomarker(s) is/are selected from the Mindin or Hacel or combinations thereof.
  • sample refers to any biological fluid, cell or tissue sample from a subject which can be assayed for biomarkers (e.g. RNA and/or polypeptide products), such as soluble biomarkers in subjects having or not having heart failure.
  • biomarkers e.g. RNA and/or polypeptide products
  • the sample is optionally or comprises blood, serum, plasma, or heart cells.
  • the sample can for example be a "post-treatment” sample wherein the sample is obtained after one or more treatments, or a "base-line sample” which is for example used as a base line for assessing disease progression.
  • the sample is a blood sample.
  • blood sample refers to any fraction or whole blood, which can comprise cells or be substantially cell free, which can be assayed for biomarkers, including for example blood, serum, and plasma.
  • antibody as used herein is intended to include monoclonal antibodies, polyclonal antibodies, and chimeric antibodies. The antibody may be from recombinant sources and/or produced in transgenic animals. Antibodies can be fragmented using conventional techniques. For example, F(ab')2 fragments can be generated by treating the antibody with pepsin. The resulting F(ab')2 fragment can be treated to reduce disulfide bridges to produce Fab' fragments. Papain digestion can lead to the formation of Fab fragments.
  • Fab, Fab' and F(ab')2, scFv, dsFv, ds- scFv, dimers, minibodies, diabodies, bispecific antibody fragments and other fragments can also be synthesized by recombinant techniques.
  • Antibodies having specificity for a specific protein may be prepared by conventional methods.
  • a mammal e.g. a mouse, hamster, or rabbit
  • an immunogenic form of the peptide which elicits an antibody response in the mammal.
  • Techniques for conferring immunogenicity on a peptide include conjugation to carriers or other techniques well known in the art.
  • the peptide can be administered in the presence of adjuvant.
  • the progress of immunization can be monitored by detection of antibody titers in plasma or serum. Standard ELISA or other immunoassay procedures can be used with the immunogen as antigen to assess the levels of antibodies.
  • antibody producing cells can be harvested from an immunized animal and fused with myeloma cells by standard somatic cell fusion procedures thus immortalizing these cells and yielding hybridoma cells.
  • myeloma cells can be harvested from an immunized animal and fused with myeloma cells by standard somatic cell fusion procedures thus immortalizing these cells and yielding hybridoma cells.
  • Hybridoma cells can be screened immunochemically for production of antibodies specifically reactive with the peptide and the monoclonal antibodies can be isolated.
  • the antibody is a purified or isolated antibody.
  • purified or isolated is meant when for example referring to an antibody, that a given antibody or fragment thereof, whether one that has been removed from nature (isolated from blood serum) or synthesized (produced by recombinant means), has been increased in purity, wherein “purity” is a relative term, not “absolute purity.”
  • a purified antibody is 60% free, preferably at least 75% free, and more preferably at least 90% free from other components with which it is naturally associated or associated following synthesis and/or during isolation.
  • biomarker specific reagent refers to any molecule or compound that can bind to a biomarker product described herein, including polypeptides such as antibodies, aptamers, nucleic acids and peptide mimetics.
  • the "biomarker specific reagent” can for example be coupled to or labeled with a detectable marker.
  • the label is preferably capable of producing, either directly or indirectly, a detectable signal.
  • the label may be radio-opaque or a radioisotope, such as 3 H, 14 C, 32 P, 35 S, 23 l, 125 l, 131 l; a fluorescent (fluorophore) or chemiluminescent (chromophore) compound, such as fluorescein isothiocyanate, rhodamine or luciferin; an enzyme, such as alkaline phosphatase, beta- galactosidase or horseradish peroxidase; an imaging agent; or a metal ion.
  • a radioisotope such as 3 H, 14 C, 32 P, 35 S, 23 l, 125 l, 131 l
  • a fluorescent (fluorophore) or chemiluminescent (chromophore) compound such as fluorescein isothiocyanate, rhodamine or luciferin
  • an enzyme such as alkaline phosphatase, beta- galactosidase
  • hybridize or “hybridizable” refers to the sequence specific non-covalent binding interaction with a complementary nucleic acid.
  • the hybridization is under high stringency conditions. Appropriate stringency conditions which promote hybridization are known to those skilled in the art, or can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1 6.3.6. For example, 6.0 X sodium chloride/sodium citrate (SSC) at about 45°C, followed by a wash of 2.0 X SSC at 50° C may be employed.
  • SSC sodium chloride/sodium citrate
  • polynucleotide refers to a sequence of nucleotide or nucleoside monomers consisting of naturally occurring bases, sugars, and intersugar (backbone) linkages, and is intended to include DNA and RNA which can be either double stranded or single stranded, represent the sense or antisense strand.
  • primer refers to a polynucleotide, whether occurring naturally as in a purified restriction digest or produced synthetically, which is capable of acting as a point of synthesis when placed under conditions in which synthesis of a primer extension product, which is complementary to a nucleic acid strand is induced (e.g. in the presence of nucleotides and an inducing agent such as DNA polymerase and at a suitable temperature and pH).
  • the primer must be sufficiently long to prime the synthesis of the desired extension product in the presence of the inducing agent.
  • the exact length of the primer will depend upon factors, including temperature, sequences of the primer and the methods used.
  • a primer typically contains 15-25 or more nucleotides, although it can contain less. The factors involved in determining the appropriate length of primer are readily known to one of ordinary skill in the art.
  • probe refers to a nucleic acid sequence that will hybridize to a nucleic acid target sequence.
  • the probe hybridizes to a biomarker RNA or a nucleic acid sequence complementary to the biomarker RNA.
  • the length of probe depends for example, on the hybridization conditions and the sequences of the probe and nucleic acid target sequence.
  • the probe can be for example, at least 15, 20, 25, 50, 75, 100, 150, 200, 250, 400, 500 or more nucleotides in length.
  • the term "specifically binds" as used herein refers to a binding reaction that is determinative of the presence of the biomarker (e.g. polypeptide or nucleic acid) often in a heterogeneous population of macromolecules.
  • the biomarker specific reagent is an antibody
  • specifically binds refers to the specified antibody binding with greater affinity to the cognate antigenic determinant than to another antigenic determinant, for example binds with at least 2, at least 3, at least 5, or at least 10 times greater specificity
  • a probe specifically binds refers to the specified probe under hybridization conditions binds to a particular gene sequence at least 1.5, at least 2 at least 3, or at least 5 times background.
  • kit control means a suitable assay control useful when determining a level of a biomarker level
  • the kit control can comprise an antibody control, useful for example for detecting non-specific binding and/or for standardizing the amount of protein in the sample.
  • the kit control can comprise an antibody that specifically binds vinculin.
  • biomarker specific reagent refers to a reagent that is a highly sensitive and specific biomarker reagent used with standard immunohistochemistry (ICC) and immunohistochemistry (IHC) techniques to detect the level of a biomarker associated with heart disease.
  • ICC immunohistochemistry
  • IHC immunohistochemistry
  • Mindin also known as DIL-1 , DILI , M-spondin and Spondin-2 precursor, refers to a Mindin gene (e.g. gene name SPON2 and DILI ) or expression products(e.g.
  • mRNA and polypeptide products including naturally occurring variants, and including mammalian Mindin, such as human Mindin, including those deposited in Genbank with accession number NM 012445, NM_001 128325, NP_001 121797, NP_036577, AAH36341 , AAH02707, 3D34_B, 3D34_A, BC002707, BC036341 , BD 88997, BD188996, AX576915, BD006593 and BD006592 and/or with UniProtKB/Swiss-Protien accession numbers Q9BUD6, Q4W5N4, Q9ULW1 (SPON2_HUMAN) and sequences included herein.
  • Mindin is an extracellular matrix protein and the gene encoding Mindin is located on chromosome 4p16.3.
  • Hacel or "HECT domain and ankyrin repeat containing, E3 ubiquitin protein ligase 1 " refers to a Hacel gene or expression products (e.g. mRNA and polypeptide products) or cDNA including naturally occurring variants, and including mammalian Hacel , such as human Hacel , including those deposited in Genbank with accession number NP 065822, NM 020771 and sequences included herein. [0058] Sequences of accession numbers referred to herein are hereby incorporated by reference.
  • heart disease refers to a variety of disorders and diseases that affect the heart includes for example ischemic heart disease, cardiomyopathy including for example dilated cardiomyopathy (DCM), cardiovascular disease, heart failure, hypertrophy, and myocardial infarction induced injuries including initial injuries during the acute adverse remodeling phase.
  • ischemic heart disease CAD
  • cardiomyopathy including for example dilated cardiomyopathy (DCM)
  • cardiovascular disease CAD
  • hypertrophy hypertrophy
  • myocardial infarction induced injuries including initial injuries during the acute adverse remodeling phase.
  • heart disease excluding myocardial infarction induced initial injuries refers to a variety of disorders and diseases that affect the heart includes for example ischemic heart disease, cardiomyopathy including for example dilated cardiomyopathy (DCM), cardiovascular disease, heart failure, hypertrophy and myocardial infarction related injuries but not including initial injuries during the acute adverse remodeling phase.
  • ischemic heart disease CAD
  • cardiomyopathy including for example dilated cardiomyopathy (DCM)
  • cardiovascular disease CAD
  • heart failure hypertrophy
  • myocardial infarction related injuries but not including initial injuries during the acute adverse remodeling phase.
  • heart failure refers to a to a clinical syndrome characterized by distinctive symptoms and signs resulting from disturbances in cardiac output or from increased venous pressure and includes chronic, acute and end-stage heart failure.
  • Heart failure includes systolic heart failure and diastolic heart failure.
  • ischemic heart disease refers to disorders characterized by reduced blood supply to the heart muscle.
  • cardiac hypertrophy refers to thickening and enlargement of the cardiac chamber, and includes established compensatory cardiac hypertrophy, and pathological decompensated cardiac hypertrophy.
  • DCM diastolic cardiomyopathy
  • Ml myocardial infarction
  • the infarcted tissue typically forms a fibrotic scar.
  • Long-term consequences include ventricular remodeling of the remaining myocardium (e.g., development of compensatory hypertrophy and/or dilation), ventricular failure, arrhythmias and sudden death.
  • Improvement in a myocardial infarction induced injury can for example be determined by detecting improvement in systolic function (e.g. ejection fraction, fractional shortening) and improvement in diastolic function as result of inhibition of adverse cardiac remodeling (decreased infarct size, decreased replacement fibrosis and fewer dying cells), inhibition of hypertrophy, inhibition of chamber dilation, and consequently inhibition of ventricular fibrillation and arrhythmias, well as for example secondary pulmonary congestion and edema.
  • systolic function e.g. ejection fraction, fractional shortening
  • diastolic function as result of inhibition of adverse cardiac remodeling (decreased infarct size, decreased replacement fibrosis and fewer
  • treatment is an approach for obtaining beneficial or desired results, including clinical results.
  • beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, reversal of disease, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable.
  • Treatment can also mean prolonging survival as compared to expected survival if not receiving treatment.
  • an "agent that inhibits a biomarker” refers to an agent that inhibits the expression or function of the biomarker and includes for example a neutralizing antibody specific for the biomarker that inhibits for example biomarker function or a siRNA molecule that inhibits the expression of the biomarker.
  • the agents can, for example injected in heart.
  • Mindin levels are increased in serum samples of subjects with ischemic heart disease and dilated cardiomyopathy. Mindin levels are increased in both ischemic and non-ischemic (e.g. dilated cardiomyopathy) heart disease, for example supporting that Mindin levels are increased in heart disease generally.
  • Hace-1 gene and protein expression levels are significantly elevated in the blood samples of systolic heart failure patients.
  • Hace-1 knockout mice also fail to develop hypertrophy following cardiac banding, suggesting a role for Hace-1 in heart disease.
  • the disclosure provides a method of screening for, diagnosing or detecting heart disease in a subject, the method comprising:
  • the method comprises:
  • the disclosure provides a method for treating a subject having heart disease comprising:
  • the method comprises: a. obtaining a sample from the subject;
  • Yet a further aspect provides a method of monitoring response to treatment comprising:
  • Another aspect provides a method of monitoring heart disease progression comprising:
  • an increase in the Mindin and/or Hacel level in the post-base-line sample compared to the base-line level is indicative the heart disease is progressing
  • a decrease in the Mindin and/or Hacel level in the post base-line sample compared to the base-line level is indicative that the heart disease is not progressing
  • the level of Mindin is determined. In an embodiment, the level of Hacel is determined.
  • the Mindin level determined is a polypeptide level.
  • the Mindin polypeptide comprises SEQ ID NO:3 and/or 5 and/or a polypeptide with at least 95%, 96%, 97%, 98% or at least 99% sequence identity with SEQ ID NO:3 and/or 5.
  • the Mindin level determined is a nucleic acid level.
  • the Mindin nucleic acid comprises SEQ ID NO:1 and/or 2 and/or a nucleic acid molecule with at least 95%, 96%, 97%, 98% or at least 99% sequence identity with SEQ ID NO: 1 and/or 2.
  • the Hace-1 level determined is a polypeptide level.
  • the Hace-1 polypeptide comprises SEQ ID NO:6 and/or a polypeptide with at least 95%, 96%, 97%, 98% or at least 99% sequence identity with SEQ ID NO:6.
  • the Hace-1 level determined is a nucleic acid level.
  • the Hace-1 nucleic acid comprises SEQ ID NO:7 and/or a nucleic acid molecule with at least 95%, 96%, 97%, 98% or at least 99% sequence identity with SEQ ID NO: 7.
  • the biomarker determined is soluble biomarker.
  • the heart disease comprises ischemic heart disease.
  • the heart disease comprises a cardiomyopathy.
  • the cardiomyopathy comprises dilated cardiomyopathy (DCM).
  • the heart disease comprises cardiovascular disease.
  • the heart disease comprises heart failure.
  • the heart disease comprises hypertrophy.
  • the heart disease comprises injuries sustained resulting from a myocardial infarction.
  • the heart disease is heart disease excluding myocardial infarction induced initial injuries.
  • the heart disease indicated comprises ischemic heart disease. In another embodiment, wherein the subject has an increased level of Mindin the heart disease indicated comprises myocardial infarction induced injury. In yet another embodiment, wherein the subject has an increased level of Mindin, the heart disease indicated comprises DCM.
  • the method is for the early detection of heart disease.
  • the method is for the early detection of ischemic heart disease.
  • it is demonstrated herein using an animal model, that Mindin shows elevation in the acute and early stage of disease (e.g. see Figure 1).
  • the heart disease is systolic heart failure.
  • the disclosure provides a method of distinguishing systolic heart failure from diastolic heart failure in a subject with heart failure, the method comprising: a. determining a level of Hacel in the sample, and
  • an increased level of Hacel in the sample compared to the control is indicative the subject has systolic heart failure and wherein a level comparable in the sample to the control is indicative the subject has diastolic heart failure.
  • the method comprises: a. determining a level of Hacel in a sample from the subject, and b. comparing the level of Hacel in the sample with a control, wherein the control is a level of Hacel in a subject without heart failure or a subject with diastolic heart failure;
  • an increased level of Hacel in the sample compared to the control is indicative the subject has systolic heart failure and wherein a level comparable in the sample to the control is indicative the subject has diastolic heart failure.
  • the level of two biomarkers are determined.
  • the method comprises obtaining a sample from the subject.
  • the sample can for example be a blood sample or a biopsy.
  • the ratio of the level of the biomarker in the sample compared to the control indicative of heart disease or a type thereof is greater than 1.2, 1.4, 1.6, 1 .8, 2, 3, 5, 10, or more. In an embodiment, the ratio is at least 1 .3, at least 1.5, at least 1.7, at least 1.9, at least 2.1 , at least 2.5 or at least 3.
  • the level of the biomarker determined is a polypeptide level or a nucleic acid level.
  • the level of Mindin is determined using a biomarker specific reagent, e.g a Mindin specific reagent such an antibody or antibody fragment, which specifically binds Mindin, for example Mindin polypeptide.
  • the Mindin specific reagent is a probe that hybridizes to Mindin mRNA or cDNA.
  • the Mindin specific reagent is a primer or primer pair for amplifying Mindin nucleic acids.
  • the level of Hacel is determined using a biomarker specific reagent e.g. a Hacel specific reagent, such an antibody or antibody fragment, which specifically binds Hacel , for example Hacel polypeptide.
  • the Hacel specific reagent is a probe that hybridizes to Hacel mRNA or cDNA.
  • the Hacel specific reagent is a primer or primer pair for amplifying Hacel nucleic acids, e.g. for example by PCR methods.
  • the level determined is a polypeptide product.
  • the step of determining the biomarker level comprises using immunohistochemistry and/or an immunoassay.
  • the immunoassay is an ELISA.
  • the ELISA is a sandwich type ELISA.
  • antibodies or antibody fragments are used to determine the level of polypeptide of one or more biomarkers of the disclosure.
  • the antibody or antibody fragment is labeled with a detectable marker.
  • the antibody or antibody fragment is, or is derived from, a monoclonal antibody.
  • a person skilled in the art will be familiar with the procedure for determining the level of a polypeptide biomarker by using said antibodies or antibody fragments, for example, by contacting the sample from the subject with an antibody or antibody fragment labeled with a detectable marker, wherein said antibody or antibody fragment forms a complex with the biomarker.
  • the label is preferably capable of producing, either directly or indirectly, a detectable signal.
  • the label may be radio-opaque or a radioisotope, such as 3 H, 14 C, 32 P, 35 S, 123 l, 125 l, 3 ; a fluorescent (fluorophore) or chemiluminescent (chromophore) compound, such as fluorescein isothiocyanate, rhodamine or luciferin; an enzyme, such as alkaline phosphatase, beta- galactosidase or horseradish peroxidase; an imaging agent; or a metal ion.
  • a radioisotope such as 3 H, 14 C, 32 P, 35 S, 123 l, 125 l, 3
  • a fluorescent (fluorophore) or chemiluminescent (chromophore) compound such as fluorescein isothiocyanate, rhodamine or luciferin
  • an enzyme such as alkaline phosphatase, beta- galactosidase or horserad
  • the level of polypeptide biomarker of the disclosure is detectable indirectly.
  • a secondary antibody that is specific for a primary antibody that is in turn specific for the isolated protein of the disclosure wherein the secondary antibody contains a detectable label can be used.
  • the Mindin specific reagent specifically binds Mindin polypeptide. In an embodiment the Mindin specific reagent specifically binds SEQ ID NO:3 and/or 5. In an embodiment, the Mindin specific reagent specifically binds Mindin nucleic acid. In an embodiment, the Mindin specific reagent specifically binds SEQ ID NO: 1 and/or 2.
  • the Hacel specific reagent specifically binds Hacel polypeptide. In an embodiment the Hacel specific reagent specifically binds SEQ ID NO:6. In an embodiment, the Hacel specific reagent specifically binds Hacel nucleic acid. In an embodiment, the Hacel specific reagent specifically binds SEQ ID NO:7.
  • the treatment is optionally an agent that inhibits the biomarker such as an antibody specific for the biomarker or a siRNA molecule that inhibits the expression of the biomarker, for example injected in heart or administered intravenously.
  • an agent that inhibits the biomarker such as an antibody specific for the biomarker or a siRNA molecule that inhibits the expression of the biomarker, for example injected in heart or administered intravenously.
  • kits for screening for, detecting, or diagnosing heart disease in a subject and/or determining prognosis of a subject having heart disease comprises one or more biomarker specific reagents, for example an antibody, specific for a biomarker described herein, for example Mindin or Hacel .
  • the kit can also include a control or reference standard e.g. such as a positive control such as Mindin or Hacel polypeptide and/or instructions for use.
  • the kit can include ancillary agents such as vessels for storing or transporting the sample for example specimen collection tubes for example for collecting a biopsy, and/or storing or transporting biomarker specific reagents and/or buffers such as extraction buffers for extracting nucleic acid, for example buffers that inhibit RNAse action or stabilizers.
  • the kit comprises an antibody to one or more of Mindin and Hacel and a quantity of a purified standard, such as a known quantity of biomarker polypeptide.
  • the disclosure provides a kit for detecting a biomarker comprising:
  • the kit comprises one or more biomarker specific reagents wherein the biomarker specific reagent binds to an extracellular portion of a biomarker for example wherein the biomarker or a fragment thereof is secreted.
  • the biomarker specific reagent is an antibody or antibody fragment.
  • the biomarker specific reagent is a probe, or primer set that amplifies a nucleic acid transcript of the biomarker.
  • the kit control is a peptide fragment of the at least one biomarker.
  • the kit comprises an ELISA plate.
  • Mindin (Spondin 2) is a highly conserved secreted extracellular matrix (ECM) protein of the Mindin-F-spondin family. Previous studies showed Mindin to be a regulator of host innate immunity. A reduced NF-kB activation and macrophage recruitment which were accompanied by a decrease in pro-apoptotic proteins (caspase 3, 8 and 9, Bad, Bax, and Bik) and an increase in pro-survival protein of MAPK (p44/42).
  • Mindin is a putative inhibitor of angiogenesis, and an increase in angiogenic factors such as vascular endothelial growth factor (VEGF) was observed post Ml.
  • VEGF vascular endothelial growth factor
  • Mindin-/- mice showed reduced NF-kB activation and macrophage recruitment, and these were accompanied by a decrease in pro-apoptotic proteins (caspase 3, 8 and 9, Bad, Bax, and Bik) and an increase in pro-survival protein of MAPK (p44/42) in Mindin-/- mice.
  • Mindin is a putative inhibitor of angiogenesis, and Mindin-/- mice did show an increase in angiogenic factors such as vascular endothelial growth factor (VEGF) post Ml.
  • VEGF vascular endothelial growth factor
  • Microarray data (Table 1 and Figure 4) has shown that Mindin is increased in heart of ischemic and dilated cardiomyopathy patients.
  • HACE1 a novel biomarker of systolic heart failure, controls cardiac hypertrophy through regulating protein degradation
  • HACE1 E3 ubiquitin-protein ligase gene
  • HACE1 acting as a novel co-chaperone / ubiquitin ligase controls myocyte protein stability by close co-regulated with heat shock proteins.
  • HACE1 A novel HECT family E3 ubiquitin-protein ligase gene, HACE1 , had previously been identified as a potential suppressor of multiple human cancers.
  • Hacel knock out mice were made. Hacel knockout mice fail to develop hypertrophy following pressure overload created by cardiac banding.
  • HACE1 acting as a novel co-chaperone / ubiquitin ligase controls myocyte protein stability by close co-regulated with heat shock proteins.
  • HACE1 could be a critical regulator of cardiac hypertrophy and remodelling.
  • HACE1 acting as a novel co- chaperone / ubiquitin ligase controls myocyte protein stability by close co-regulated with heat shock proteins.
  • GO biological process ubiquitin cycle
  • GO molecular function ubiquitin-protein ligase activity
  • spondin 2 extracellular matrix protein (SPON2)
  • transcript variant 2 mRNA
  • SPON2 Homo sapiens spondin 2, extracellular matrix protein
  • transcript variant 1 mRNA
  • SEQ ID NO:2 1 gataggacag acagacaaag aaaggggtgc ggcagcactg ccaggggaag agggtgatcc
  • mouse mindin protein which is 85% identical to the human protein.
  • extracellular matrix protein [Mus muscu 330 aa SEQ ID NO:4 1 menvslalgr alwvfllami gsttsqplgg esvctarpla rysitfigkw sqtafpkqyp
  • HACE1 ubiquitin protein ligase 1
  • LOCUS NP_766061 909 aa linear 10-FEB-2008 HECT domain and ankyrin repeat containing, E3 ubiquitin protein

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

L'invention porte sur des biomarqueurs permettant de détecter une cardiopathie, qui comprennent des biomarqueurs détectables dans le sang. L'invention porte sur une méthode pour le criblage, le diagnostic ou la détection d'une cardiopathie chez un sujet, la méthode consistant : a. à déterminer un niveau de Mindin et/ou de Hace dans l'échantillon; et b. à comparer le niveau de Mindin et/ou de Hace dans l'échantillon à un témoin; un niveau élevé de Mindin et/ou de Hace dans l'échantillon comparé au témoin indiquant que le sujet souffre d'une cardiopathie.
PCT/CA2010/001426 2009-09-11 2010-09-13 Méthodes et kits pour la détection de cardiopathie Ceased WO2011029201A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US24164809P 2009-09-11 2009-09-11
US61/241,648 2009-09-11

Publications (1)

Publication Number Publication Date
WO2011029201A1 true WO2011029201A1 (fr) 2011-03-17

Family

ID=43731900

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CA2010/001426 Ceased WO2011029201A1 (fr) 2009-09-11 2010-09-13 Méthodes et kits pour la détection de cardiopathie

Country Status (1)

Country Link
WO (1) WO2011029201A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040072227A1 (en) * 1997-05-09 2004-04-15 Jonak Zdenka L. Integrin ligand, human mindin
US20070031847A1 (en) * 2003-03-10 2007-02-08 Applera Corporation Genetic polymorphisms associated with stenosis, methods of detection and uses thereof
WO2008003826A1 (fr) * 2006-07-07 2008-01-10 Oy Jurilab Ltd Nouveaux gènes et marqueurs dans l'hypertension arterielle essentielle

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040072227A1 (en) * 1997-05-09 2004-04-15 Jonak Zdenka L. Integrin ligand, human mindin
US20070031847A1 (en) * 2003-03-10 2007-02-08 Applera Corporation Genetic polymorphisms associated with stenosis, methods of detection and uses thereof
WO2008003826A1 (fr) * 2006-07-07 2008-01-10 Oy Jurilab Ltd Nouveaux gènes et marqueurs dans l'hypertension arterielle essentielle

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
MOON, M.S. ET AL., CIRCULATION, vol. 120, 3 November 2009 (2009-11-03), pages S791 *
PARRY, R. ET AL., CANCER RES., vol. 65, no. 18, 15 September 2005 (2005-09-15), pages 8397 - 8405 *

Similar Documents

Publication Publication Date Title
EP2260301B9 (fr) Signatures géniques
US20110053170A1 (en) Interleukin-33 (IL-33) for the Diagnosis and Prognosis of Cardiovascular Disease
EP3710831A1 (fr) Marqueurs pour le diagnostic et le traitement de la stéatohépatite non alcoolique (nash) et de la fibrose hépatique avancée
JP2016218084A (ja) 心血管疾患の診断方法
JP2011209291A (ja) アルツハイマー病に関する方法及び組成物
WO2016152352A1 (fr) Biomarqueur spécifique du mélanome et son utilisation
WO2006034356A9 (fr) Genes associes a la surcharge de pression cardiaque
JP5568807B2 (ja) プロテオミクス解析を用いたメラノーママーカーの同定
KR102547531B1 (ko) 주요 우울 장애 치료 반응성 평가용 바이오마커
JP2025518766A (ja) 胃癌発病可能性の予測のためのバイオマーカー及びその用途
KR102015982B1 (ko) 고밀도 지단백 기능과 관련된 심혈관 질환 치료 표적 및 이를 이용하여 심혈관 질환을 진단하는 방법
Xie et al. Clinical significance of MiR-27a expression in serum exosomes in patients with heart failure
JP7177906B2 (ja) 心筋損傷の診断方法
WO2011029201A1 (fr) Méthodes et kits pour la détection de cardiopathie
EP3390669A1 (fr) Biomarqueurs de la leucémie lymphoïde chronique et leur utilisation
JP5885243B2 (ja) 胆管細胞癌の検出方法および予防・治療剤のスクリーニング方法
KR102026252B1 (ko) 폐 편평상피세포암 바이오마커 및 이를 이용한 폐 편평상피세포암 진단방법
WO2020058701A1 (fr) Méthode d'identification d'une cardiomyopathie hypertrophique
JP7239139B2 (ja) 肝硬変の診断方法、非アルコール性脂肪肝炎及び肝細胞がんの合併症の診断方法並びに非アルコール性脂肪肝炎及び食道胃静脈瘤の合併症の診断方法
JP7389108B2 (ja) 心房細動関連脳卒中の評価のためのｃｅｓ－２（カルボキシルエステラーゼ－２）
JP2004321179A (ja) 炎症性腸疾患診断用マーカー遺伝子もしくは蛋白質およびそれを利用した炎症性腸疾患診断方法
KR102488525B1 (ko) 비흡연성 비소세포 폐암 환자의 면역상태 판단용 바이오마커 및 이를 이용한 비흡연성 비소세포 폐암 환자의 면역 상태에 대한 정보를 제공하는 방법
WO2023027186A1 (fr) Biomarqueur de fibrose pulmonaire
JP2004173553A (ja) 心不全の疾患マーカー及びその利用
JP2004129587A (ja) 中性脂質の蓄積を伴う疾患の疾患マーカー及びその利用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 10814858

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 10814858

Country of ref document: EP

Kind code of ref document: A1