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WO2008112424A1 - Évaluation de l'insuffisance cardiaque - Google Patents

Évaluation de l'insuffisance cardiaque Download PDF

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Publication number
WO2008112424A1
WO2008112424A1 PCT/US2008/055019 US2008055019W WO2008112424A1 WO 2008112424 A1 WO2008112424 A1 WO 2008112424A1 US 2008055019 W US2008055019 W US 2008055019W WO 2008112424 A1 WO2008112424 A1 WO 2008112424A1
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Prior art keywords
polypeptide
cofilin
level
mammal
cardiovascular condition
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PCT/US2008/055019
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English (en)
Inventor
Jr. John C. Burnett
Brenda K. Huntley
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Mayo Foundation for Medical Education and Research
Mayo Clinic in Florida
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Mayo Foundation for Medical Education and Research
Mayo Clinic in Florida
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    • 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
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
    • G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
    • G01N2333/4701—Details
    • G01N2333/4712—Muscle proteins, e.g. myosin, actin, protein
    • 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
    • G01N2800/325—Heart failure or cardiac arrest, e.g. cardiomyopathy, congestive heart failure

Definitions

  • This document relates to methods and materials involved in assessing cardiovascular conditions (e.g., heart failure) in mammals and assessing treatments thereof.
  • cardiovascular conditions e.g., heart failure
  • this document relates to methods and materials involved in using a cofilin-1 polypeptide as a marker to determine whether or not a mammal (e.g., a human) has a cardiovascular condition.
  • Heart failure also referred to as congestive heart failure or congestive cardiac failure, is a condition in which the heart cannot pump enough blood to meet the body's need for oxygen and nutrients, which are supplied by the blood.
  • the pumping action of the heart is inadequate, blood flow to body tissues can be reduced, and blood returning to the heart can accumulate and cause congestion in the veins.
  • Accumulation of blood coming into the left side of the heart can cause congestion in the lungs and impair lung function.
  • Accumulation of blood coming into the right side of the heart from the rest of the body can cause congestion in other parts of the body, such as the legs and liver.
  • Heart failure usually affects both the right and left sides of the heart to some degree. However, one side may be affected more than the other.
  • Heart failure can occur in people of any age, it is more common among elderly. Heart failure develops in about one out of 100 people and is likely to become more common because people are living longer and because certain risk factors for heart disease, such as smoking, high blood pressure, and a high-fat diet, are affecting more people.
  • This document provides methods and materials related to assessing cardiovascular conditions and treatments of cardiovascular conditions. For example, this document provides methods and materials for determining whether or not a mammal (e.g., a human) has a cardiovascular condition (e.g., heart failure), methods and materials for determining whether or not a cardiovascular condition is progressing, and methods and materials for determining whether or not a treatment for a cardiovascular condition is effective.
  • a cofilin-1 polypeptide e.g., the level of a cofilin-1 polypeptide in a plasma sample from a mammal
  • the level of a cofilin-1 polypeptide can indicate the state (e.g., a mild, moderate, or severe state) of a cardiovascular condition.
  • the level of a cofilin-1 polypeptide can be monitored (e.g., over time or in response to a treatment for a cardiovascular condition) to determine whether or not the state of a cardiovascular condition has changed. Having the ability to identify and monitor cardiovascular conditions can allow clinicians to diagnose cardiovascular conditions at earlier stages, to monitor progression of cardiovascular conditions effectively, and to determine proficiently the effectiveness of treatments for cardiovascular conditions.
  • an antibody directed against a cofilin-1 polypeptide can be used to treat a mammal having a cardiovascular condition (e.g., heart failure). Having the ability to treat cardiovascular conditions can allow clinicians to reduce the significant death and disability associated with cardiovascular conditions.
  • one aspect of this document features a method for identifying a cardiovascular condition.
  • the method comprises, or consists essentially of, (a) determining whether or not a plasma sample from a mammal contains an elevated level of a cofilin-1 polypeptide, and (b) classifying the mammal as having a cardiovascular condition if the plasma sample contains the elevated level and classifying the mammal as not having a cardiovascular condition if the plasma sample does not contain the elevated level.
  • the mammal can be a human.
  • An antibody directed against the cofilin-1 polypeptide can be used in the determining step.
  • the cofilin-1 polypeptide can be a human cofilin-1 polypeptide.
  • the cardiovascular condition can be heart failure.
  • the elevated level can be a detectable level of the cofilin-1 polypeptide.
  • the elevated level can be a level greater than 50 ng of the cofilin-1 polypeptide per mL of the plasma sample.
  • the elevated level can be a level greater than a reference level.
  • this document features a method for assessing the effectiveness of a treatment for a cardiovascular condition in a mammal.
  • the method comprises, or consists essentially of, determining whether or not a mammal having a cardiovascular condition and having received a treatment for the cardiovascular condition has a plasma level of a cof ⁇ lin-1 polypeptide that is the same or greater than that present prior to the treatment, wherein the presence of the plasma level that is the same or greater than that present prior to the treatment indicates that the treatment is not effective.
  • this document features a method for monitoring a cardiovascular condition in a mammal.
  • the method comprises, or consists essentially of, determining whether or not the level of a cof ⁇ lin-1 polypeptide in a first sample from a mammal having a cardiovascular condition is greater than the level of the cofilin-1 polypeptide in a second sample from the mammal, wherein the second sample is from the mammal at a time later than that of the first sample, wherein the determining step indicates that the cardiovascular condition has improved when the level of the cofilin-1 polypeptide in the first sample is greater than the level of the cofilin-1 polypeptide in the second sample.
  • this document features a method for treating a cardiovascular condition in a mammal.
  • the method comprises, or consists essentially of, administering an anti-cofilin-1 polypeptide antibody to the mammal under conditions wherein the severity of a symptom of the cardiovascular condition is reduced.
  • Figure 1 is an image of plasma polypeptides from normal subjects and heart failure patients having medium or high plasma levels of BNP polypeptides. The polypeptides were separated on a polyacrylamide gel and stained with Coomassie blue.
  • Figure 2 is a Western blot analyzing non-phosphorylated and phosphorylated cofilin-1 polypeptide levels in plasma samples from normal subjects and heart failure patients having medium or high plasma levels of BNP polypeptides.
  • Figure 3 contains graphs plotting intensity of cof ⁇ lin-1 nucleic acid expression in left ventricle (LV) or left atrium (LA) samples from a rat model of nephrectomy (NX), myocardial infarction (MI), and combined nephrectomy with MI (NM), or from a canine model of ALVD or PLVD.
  • LV left ventricle
  • LA left atrium
  • Figure 4 contains Western blots analyzing expression of unphosphorylated and phosphorylated cofilin-1 polypeptides in cytoplasmic and nuclear fractions of human cardiac fibroblasts (HCF) that were or were not plated on fibronectin coated plates, and that were either mock-treated or treated with BNP polypeptide.
  • Cells plated on fibronectin have increased phophorylation of cofilin-1 polypeptides.
  • BNP polypeptide treatment of cells plated on fibronectin inhibited or decreased this increase in phosphorylation.
  • Figure 5 contains photomicrographs of tissue sections from normal or ALVD dogs that were analyzed for cofilin-1 polypeptide expression by immunohistochemistry.
  • RV right ventricle
  • LV left ventricle
  • RA right atrium
  • LA left atrium.
  • the Immunohistochemistry results show decreased expression of cofilin-1 polypeptide in canine heart failure tissue.
  • the pattern is the same as the natriuretic peptides which show decreased tissue expression in heart failure tissue, while becoming highly expressed in the plasma-released from tissue into the plasma in heart failure.
  • Figure 6 is a graph plotting the amounts of cofilin-1 polypeptide in urine samples from patients at the indicated stages of heart failure, as measured using an ELISA.
  • This document provides methods and materials related to assessing cardiovascular conditions and treatments for cardiovascular conditions in mammals. For example, this document provides methods and materials for determining whether or not a sample (e.g., a plasma sample) from a mammal contains a cof ⁇ lin-1 polypeptide. As disclosed herein, if a sample from a mammal is observed to contain a cof ⁇ lin-1 polypeptide, then the mammal can be classified as having a cardiovascular condition. If a sample from a mammal is not observed to contain a cof ⁇ lin-1 polypeptide, then the mammal can be classified as not having a cardiovascular condition.
  • a sample e.g., a plasma sample
  • a cof ⁇ lin-1 polypeptide can be any polypeptide having cof ⁇ lin-1 polypeptide activity including, without limitation, native and mutant cofilin-1 polypeptides.
  • a cofilin-1 polypeptide can be a human cofilin-1 polypeptide having one or more amino acid changes.
  • a cofilin-1 polypeptide can be a homo log or an ortholog of a cofilin-1 polypeptide.
  • Examples of cofilin-1 polypeptides can include, without limitation, cofilin-1 polypeptides set forth in GenBank ® gi number 5031635, 57164155, 75052662, or 54035753.
  • an elevated level as used herein with respect to the level of a cofilin-1 polypeptide is any level that is greater than a control cofilin-1 polypeptide level associated with normal, healthy mammals lacking a cardiovascular condition. In some cases, an elevated level of a cofilin-1 polypeptide can be a detectable level. In some cases, an elevated level of a cofilin-1 polypeptide can be any level that is greater than a reference level for a cofilin-1 polypeptide.
  • reference level as used herein with respect to a cofilin-1 polypeptide level is the level of a cofilin-1 polypeptide typically found in mammals free of cardiovascular conditions.
  • a reference level of a cofilin-1 polypeptide can be the average level of cofilin-1 polypeptide that is present in samples obtained from a random sampling of 50 healthy mammals. It will be appreciated that levels from comparable samples are used when determining whether or not a particular level is an elevated level.
  • An elevated level of a cofilin-1 polypeptide can be any level provided that the level is greater than a corresponding reference level for a cofilin-1 polypeptide.
  • an elevated level of a cofilin-1 polypeptide can be 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or more times greater than the reference level for a cofilin-1 polypeptide.
  • a reference level can be any amount.
  • a reference level for a cofilin-1 polypeptide can be zero. In this case, any level of cofilin-1 polypeptide greater than zero would be an elevated level.
  • any method can be used to determine the level of a cof ⁇ lin-1 polypeptide present within a sample.
  • anti-cof ⁇ lin-1 polypeptide antibodies can be used to determine the level of cofilin-1 polypeptide expression within a sample.
  • the level of a cof ⁇ lin-1 polypeptide present within a sample can be determined using polypeptide detection methods such as Western blot, ELISA, and immunochemistry techniques.
  • the level of a cof ⁇ lin-1 polypeptide present within a sample also can be determined by measuring the level of an mRNA that encodes a cofilin-1 polypeptide. Any method can be used to measure the level of an RNA encoding a cofilin-1 polypeptide including, without limitation, PCR-based methods.
  • any type of sample can be used to evaluate the level of a cofilin-1 polypeptide including, without limitation, serum, blood, and plasma.
  • any method can be used to obtain a sample.
  • a blood sample can be obtained by peripheral venipuncture. Once obtained, a sample can be manipulated prior to measuring the level of a cofilin-1 polypeptide.
  • a blood sample can be treated such that total mRNA is obtained. Once obtained, the total mRNA can be evaluated to determine the level of cofilin-1 mRNA present.
  • a blood sample can be disrupted to obtain a cell lysate. Once obtained, the cell lysate can be analyzed using anti-cofilin-1 polypeptide antibodies to determine the level of cofilin-1 polypeptide present within the sample.
  • This document also provides methods and materials to assist medical or research professionals in determining whether or not a mammal has a cardiovascular conditions.
  • Medical professionals can be, for example, doctors, nurses, medical laboratory technologists, and pharmacists.
  • Research professionals can be, for example, principle investigators, research technicians, postdoctoral trainees, and graduate students.
  • a professional can be assisted by (1) determining the level of a cofilin-1 polypeptide in a sample, and (2) communicating information about that level to that professional.
  • Any method can be used to communicate information to another person (e.g., a professional).
  • information can be given directly or indirectly to a professional.
  • any type of communication can be used to communicate the information.
  • mail, e-mail, telephone, and face-to-face interactions can be used.
  • the information also can be communicated to a professional by making that information electronically available to the professional.
  • the information can be communicated to a professional by placing the information on a computer database such that the professional can access the information.
  • the information can be communicated to a hospital, clinic, or research facility serving as an agent for the professional.
  • Methods and materials for treating a cardiovascular condition in a mammal also are provided.
  • methods and materials for using an anti-cof ⁇ lin-1 polypeptide antibody to treat a cardiovascular condition are provided.
  • Mammals identified as having a cardiovascular condition or being susceptible to develop a cardiovascular condition can be treated by administering an anti-cofilin-1 polypeptide antibody.
  • Administering an anti-cofilin-1 polypeptide antibody to a mammal can reduce the severity or frequency of one or more symptoms of the cardiovascular condition in the mammal.
  • administering an anti-cofilin-1 polypeptide antibody to a mammal can reduce the severity or frequency of fluid accumulation and swelling (edema) in the feet, ankles, legs, liver, and abdomen; nausea or loss of appetite, weight, or muscle due to fluid accumulation in the liver or stomach; fluid accumulation in the lungs; shortness of breath; acute pulmonary edema; difficulty breathing; rapid breathing; bluish skin; feelings of restlessness, anxiety, or suffocation; spasms of the airways (bronchospasms); wheezing; Cheyne-Stokes respiration (periodic breathing); formation of blood clots; or stroke.
  • edema fluid accumulation and swelling
  • fluid accumulation in the lungs shortness of breath
  • acute pulmonary edema difficulty breathing
  • rapid breathing bluish skin
  • feelings of restlessness, anxiety, or suffocation spasms of the airways (bronchospasm
  • administering an anti-cofilin-1 polypeptide antibody to a mammal can reduce the severity or frequency of chest pain, heart rhythms, heart attacks, leg cramps, high blood pressure, or kidney failure.
  • the effect of administering an anti-cofilin-1 polypeptide antibody on a symptom of a cardiovascular condition can be of any magnitude.
  • administering an anti-cofilin-1 polypeptide antibody can reduce the severity or frequency of a symptom of a cardiovascular condition by 1%, 2%, 3%, 4%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more.
  • An antibody can be, without limitation, a polyclonal, monoclonal, human, humanized, chimeric, or single-chain antibody, or an antibody fragment having binding activity, such as a Fab fragment, F(ab') fragment, Fd fragment, fragment produced by a Fab expression library, fragment comprising a VL or VH domain, or epitope binding fragment of any of the above.
  • An antibody can be of any type, (e.g., IgG, IgM, IgD, IgA or IgY), class (e.g., IgGl, IgG4, or IgA2), or subclass.
  • an antibody can be from any animal including birds and mammals. For example, an antibody can be a human, rabbit, sheep, or goat antibody.
  • An antibody can be naturally occurring, recombinant, or synthetic. Antibodies can be generated and purified using any suitable methods known in the art. For example, monoclonal antibodies can be prepared using hybridoma, recombinant, or phage display technology, or a combination of such techniques. In some cases, antibody fragments can be produced synthetically or recombinantly from a gene encoding the partial antibody sequence.
  • An anti-cofilin-1 polypeptide antibody can bind to cofilin-1 polypeptides at an affinity of at least 10 4 mol 1 (e.g., at least 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , or 10 12 mol "1 ).
  • An anti-cofilin-1 polypeptide antibody can be administered to a mammal in any amount, at any frequency, and for any duration effective to achieve a desired outcome.
  • an anti-cofilin-1 polypeptide antibody can be administered to a mammal under conditions where one or more symptoms of a cardiovascular condition (e.g., one or more symptoms described herein) are prevented or reduced.
  • An effective amount of an anti-cofilin-1 polypeptide antibody can be any amount that reduces the severity of a cardiovascular condition without producing significant toxicity to the mammal. If a particular mammal fails to respond to a particular amount, then the amount can be increased by, for example, two-fold. After receiving this higher dose, the mammal can be monitored for both responsiveness to the treatment and toxicity symptoms, and adjustments made accordingly. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, and route of administration may require an increase or decrease in the actual effective amount administered.
  • the frequency of administration can be any frequency that reduces the severity of a cardiovascular condition without producing significant toxicity to the mammal.
  • the frequency of administration can be from about four times a day to about once a week, or from about once a day to about once a month, or from about once every other day to about once a year.
  • the frequency of administration can remain constant or can be variable during the duration of treatment.
  • various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, and route of administration may require an increase or decrease in administration frequency.
  • An effective duration for administering an anti-cofilin-1 polypeptide antibody can be any duration that reduces the severity of a cardiovascular condition without producing significant toxicity to the mammal.
  • an effective duration can vary from several days to several weeks, months or years. Multiple factors can influence the actual effective duration for administering an anti-cofilin-1 polypeptide antibody.
  • an effective duration can vary with the frequency of administration, effective amount, and route of administration.
  • compositions containing an anti-cofilin-1 polypeptide antibody can be admixed, encapsulated, conjugated, or otherwise associated with other molecules to assist in uptake, distribution, and/or absorption.
  • compositions containing an anti-cofilin-1 polypeptide antibody can contain one or more pharmaceutically acceptable carriers.
  • a pharmaceutically acceptable carrier is a pharmaceutically acceptable solvent, suspending agent, or any other pharmacologically inert vehicle.
  • An anti-cofilin-1 polypeptide antibody can be administered by a number of methods depending on whether local or systemic treatment is desired and upon the area to be treated.
  • Administration can be, for example, pulmonary (e.g., by inhalation or insufflation of powders or aerosols); oral; or parenteral (e.g., by subcutaneous, intrathecal, intraventricular, intramuscular, or intraperitoneal injection, or by intravenous drip).
  • pulmonary e.g., by inhalation or insufflation of powders or aerosols
  • parenteral e.g., by subcutaneous, intrathecal, intraventricular, intramuscular, or intraperitoneal injection, or by intravenous drip.
  • Plasma samples were obtained from normal subjects and heart failure patients.
  • the SwellGel Blue Albumin Removal Kit (Pierce, Rockford, IL) was used according to the manufacturer's instructions to deplete the plasma samples of albumin and facilitate detection of other polypeptides in the plasma. Briefly, SwellGel Blue Discs were rehydrated with ultrapure water, and 100 ⁇ L of plasma were added per disc. Albumin bound to the resin and other polypeptides was removed by four successive washes with Binding/Wash Buffer.
  • the membranes were blocked to reduce non-specific background and incubated with primary antibodies to cofilin (1 :10,000; Abeam, Cambridge, MA) or cofilin-phosphoS3 (1 :1000, Abeam), for one hour at room temperature.
  • the membranes were washed in Tween-20 buffer and incubated with goat anti-rabbit horseradish peroxidase (1 : 10,000) for one hour at room temperature.
  • the immunoreactive bands were visualized by ECL Plus detection (Amersham, Piscataway, NJ) and exposed to X-ray film.
  • Coomassie Blue-stained polypeptide bands in the SDS- PAGE gel were prepared for analysis by mass spectrometry using the following procedures.
  • the gel bands were destained with a solution containing 50 mM Tris, pH 8.1, and 50% acetonitrile until most of the blue color left the gel bands.
  • the bands were then reduced with a solution containing 20 mM DTT and 5OmM Tris, pH 8.1, at 55 0 C for 40 minutes, and alkylated with 40 mM iodoacetamide at room temperature for 40 minutes in the dark.
  • Polypeptides were digested in-situ with 30 ⁇ L (0.004 ⁇ g/ ⁇ L) trypsin (Promega Corporation, Madison WI) in a solution containing 20 mM Tris, pH 8.1, and 0.0002% Zwittergent 3-16, at 37 0 C overnight followed by peptide extraction with 60 ⁇ L of 2% trifluoroacetic acid, and then 60 ⁇ L of acetonitrile.
  • trypsin Promega Corporation, Madison WI
  • the pooled extracts were concentrated to less than five ⁇ L using a SpeedVac spinning concentrator (Savant Instruments, Holbrook NY) and then diluted in 0.1% formic acid for polypeptide identification by nano-flow liquid chromatography electrospray tandem mass spectrometry (nanoLC-ESI-MS/MS) using a ThermoFinnigan LTQ Orbitrap Hybrid Mass Spectrometer 1 (ThermoElectron, Bremen, Germany) coupled to an Eksigent nanoLC-2D HPLC system (Eksigent, Dublin, CA).
  • the polypeptide mixture was loaded onto a 250nl OPTI-PAK trap (Optimize Technologies, Oregon City, OR) custom packed with Michrom Magic C8 solid phase (Michrom Bioresources, Auburn, CA) and eluted with a 0.2% formic acid/acetonitrile gradient through a Michrom packed tip capillary Magic C18 column (75 ⁇ m x 150 mm).
  • the LTQ Orbitrap mass spectrometer experiment was set to perform a FT full scan from 380-1600 m/z with resolving power set at 60000 (400m/z), followed by linear ion trap MS/MS scans on the top 3 ions. Dynamic exclusion was set to 2 and selected ions were placed on an exclusion list for 60 seconds.
  • the MS/MS raw data were converted to DTA files using ThermoElectron Bio works 3.2 and correlated to theoretical fragmentation patterns of tryptic peptide sequences from the Swissprot databases using both SEQUESTTM (ThermoElectron, San Jose, CA) and MascotTM (Matrix Sciences London, UK) search algorithms running on 10 node cluster. All searches were conducted with fixed cysteine modifications of +57 for carboxamidomethyl-cysteines and variable modifications allowing +16 with methionines for methione sulphoxide, and +42 for protein N-terminal acetylation. The search was restricted to trypsin generated peptides allowing for 2 missed cleavages and was left open to all species.
  • Peptide mass search tolerances were set to 10 ppm and fragment mass tolerance was set to ⁇ 0.8 Daltons.
  • Polypeptide identifications were considered when both Mascot and Sequest gave at least two consensus peptides with individual cross correlation or probability scores exceeding a threshold dependent on the precursor charge state, and ranking number one of all the hits for their respective MS/MS spectra.
  • a polypeptide band of about 20 kDa was visualized on Coomassie blue stained gels in plasma samples from patients having medium and high plasma levels of BNP polypeptides ( Figure 1).
  • the polypeptide band was excised and sequenced. Using database analysis, the sequence was identified as cofilin-1 polypeptide.
  • cofilin-1 polypeptide was analyzed for cofilin-1 polypeptide by Western blotting. Expression of cofilin-1 polypeptide was observed to be weak in plasma samples from patients having medium plasma BNP polypeptide levels, and higher in plasma samples from patients having high plasma BNP polypeptide levels ( Figure 2A).
  • Example 2 Analyzing gene expression in animal models of heart failure Microarray technology was used to profile gene expression in left ventricular tissue from a rat model of nephrectomy, myocardial infarction (MI), and combined nephrectomy with MI (Van Dokkum et al, J. Am. Soc. Nephrol., 15:3103-3110 (2004)).
  • Microarray technology also was used to profile gene expression in left ventricular and left atrial tissues from a canine model of asymptomatic left ventricular dysfunction (ALVD; Martin et al., Kidney Int., 67(5): 1723-30 (2005)), and a canine model of postinfarction left ventricular dysfunction (PLVD; Costello-Boerrigter et al., J. Card. Fail., 9(5), SuppL, Sl :003 (2003)).
  • ALVD canines were or were not treated with DOCA (1000 ⁇ g/kg injection).
  • Corresponding tissues from control animals also were analyzed. Left ventricular and left atrial tissues were snap frozen immediately after they were harvested from the animals, and total RNA was extracted from each specimen. Each RNA sample was linearly amplified, labeled, and hybridized to an array (Affymetrix, Santa Clara, CA). The arrays were washed, stained, and scanned.
  • cofilin-1 nucleic acid was up-regulated in left ventricular tissues and particularly in left atrial tissues from the canine model of ALVD as well as the canine model of PLVS ( Figure 3). Expression of cofilin-1 nucleic acid also was up-regulated in left ventricular tissues from the rat models of nephrectomy, myocardial infarction (MI), and combined nephrectomy with MI ( Figure 3).
  • Example 3 Measuring cofilin-1 polypeptide levels in plasma Plasma samples were obtained from normal subjects and from patients at various stages of heart failure (stage 1, stage 2, stage 3, and stage 4). The amount of cofilin-1 polypeptide in each plasma sample was measured using an ELISA with anti-cofilin-1 antibodies (Abeam, Cambridge, MA). A trend was observed toward increasing plasma cofilin-1 polypeptide level with increasing stage of heart failure (Figure 6). Three of the measurements of plasma cofilin-1 polypeptide levels in normal subjects were observed to be outliers ( Figure 6). One of the samples (corresponding to the middle outlier data point) was obtained from a young overweight subject having a pre-gastric bypass procedure. Another one of the samples (corresponding to the upper outlier data point) was obtained from an older male).
  • Urine samples also were analyzed for cofilin-1 polypeptide levels using the ELISA. Cofilin-1 polypeptide was not detected in any of the urine samples ( Figure 6).
  • a more sensitive assay will be developed to measure plasma cofilin-1 polypeptide levels. The assay will be performed using an anti-cofilin-1 antibody selected for ELISA. The assay will be used to measure cofilin-1 polypeptide levels in large numbers of plasma samples (e.g., about 100 or more plasma samples) from normal subjects and heart failure patients at various stages of failure.
  • HCF Human cardiac fibroblasts

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Abstract

Cette invention se rapporte à des procédés et des matériaux associés à l'identification, à la surveillace et au traitement des affections cardiovasculaires chez le mammifère. L'invention concerne, par exemple, des procédés et des matériaux permettant de déterminer si un mammifère (par exemple l'homme) est atteint ou non d'une affection cardiovasculaire (par exemple l'insuffisance cardiaque), des procédés et des matériaux permettant de déterminer si une affection cardiovasculaire évolue ou non, ainsi que des procédés et des matériaux permettant de déterminer si le traitement d'une affection cardiovasculaire est efficace ou non.
PCT/US2008/055019 2007-03-08 2008-02-26 Évaluation de l'insuffisance cardiaque Ceased WO2008112424A1 (fr)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012072683A3 (fr) * 2010-11-30 2012-09-07 Inserm (Institut National De La Sante Et De La Recherche Medicale) Diagnostic de dysfonctionnement systolique ventriculaire gauche asymptomatique
CN102782499A (zh) * 2010-03-03 2012-11-14 东丽株式会社 胃癌检测用的标志物和胃癌检测方法
EP2650307A4 (fr) * 2010-12-09 2014-09-17 Toray Industries PROCÉDÉ DE MESURE IMMUNOLOGIQUE DE LA PROTÉINE cofiline-1
EP3041514A4 (fr) * 2013-09-03 2017-07-05 Mayo Foundation for Medical Education and Research Réduction du risque d'événements cardiaques négatifs majeurs

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040197930A1 (en) * 2003-03-25 2004-10-07 Ron Rosenfeld Proteomic analysis of biological fluids
WO2004084703A2 (fr) * 2003-03-24 2004-10-07 Biosphere Medical, Inc. Embolisation temporaire effectuee a l'aide de polymeres thermosensibles reversibles a action momentanee

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004084703A2 (fr) * 2003-03-24 2004-10-07 Biosphere Medical, Inc. Embolisation temporaire effectuee a l'aide de polymeres thermosensibles reversibles a action momentanee
US20040197930A1 (en) * 2003-03-25 2004-10-07 Ron Rosenfeld Proteomic analysis of biological fluids

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102782499B (zh) * 2010-03-03 2015-06-10 东丽株式会社 胃癌检测用的标志物和胃癌检测方法
CN102782499A (zh) * 2010-03-03 2012-11-14 东丽株式会社 胃癌检测用的标志物和胃癌检测方法
US20130011865A1 (en) * 2010-03-03 2013-01-10 Michimoto Kobayashi Marker for detecting gastric cancer and method for detecting gastric cancer
EP2544004A4 (fr) * 2010-03-03 2013-07-31 Toray Industries Marqueur de cancer gastrique et méthode de détection de cancer gastrique
WO2012072683A3 (fr) * 2010-11-30 2012-09-07 Inserm (Institut National De La Sante Et De La Recherche Medicale) Diagnostic de dysfonctionnement systolique ventriculaire gauche asymptomatique
US9127054B2 (en) 2010-12-09 2015-09-08 Toray Industries, Inc. Immunoassay of cofilin 1 protein
EP2650307A4 (fr) * 2010-12-09 2014-09-17 Toray Industries PROCÉDÉ DE MESURE IMMUNOLOGIQUE DE LA PROTÉINE cofiline-1
EP3041514A4 (fr) * 2013-09-03 2017-07-05 Mayo Foundation for Medical Education and Research Réduction du risque d'événements cardiaques négatifs majeurs
US9884090B2 (en) 2013-09-03 2018-02-06 Mayo Foundation For Medical Education And Research Using nucleic acids encoding NAP-2 and TGF-alpha polypeptides to improve cardiac function
EP3639860A1 (fr) * 2013-09-03 2020-04-22 Mayo Foundation for Medical Education and Research Réduction du risque de problèmes cardiaques majeurs
US10682394B2 (en) 2013-09-03 2020-06-16 Mayo Foundation For Medical Education And Research NAP-2 polypeptides and methods for modulating immune system activity in heart tissue
US11413330B2 (en) 2013-09-03 2022-08-16 Mayo Foundation For Medical Education And Research Using nucleic acids encoding NAP-2 polypeptides to improve cardiac function
US12357676B2 (en) 2013-09-03 2025-07-15 Mayo Foundation For Medical Education And Research Method of using NAP-2 and TGF-α to improve cardiac function

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