[go: up one dir, main page]

WO2017061809A1 - Composition destinée au diagnostic du dysfonctionnement de cellules endothéliales vasculaires, et son utilisation - Google Patents

Composition destinée au diagnostic du dysfonctionnement de cellules endothéliales vasculaires, et son utilisation Download PDF

Info

Publication number
WO2017061809A1
WO2017061809A1 PCT/KR2016/011223 KR2016011223W WO2017061809A1 WO 2017061809 A1 WO2017061809 A1 WO 2017061809A1 KR 2016011223 W KR2016011223 W KR 2016011223W WO 2017061809 A1 WO2017061809 A1 WO 2017061809A1
Authority
WO
WIPO (PCT)
Prior art keywords
vascular endothelial
protein
endothelial cells
channel
level
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/KR2016/011223
Other languages
English (en)
Korean (ko)
Inventor
서석효
최신규
김지애
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.)
Ewha Womans University
Original Assignee
Ewha Womans University
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 Ewha Womans University filed Critical Ewha Womans University
Publication of WO2017061809A1 publication Critical patent/WO2017061809A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • 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

Definitions

  • the present invention relates to a composition for diagnosing vascular endothelial dysfunction and its use. More specifically, the present invention relates to a composition for diagnosing vascular endothelial dysfunction, a kit, a method for diagnosing vascular endothelial dysfunction, and vascular endothelial dysfunction. The present invention relates to a method for screening an induction substance for improvement.
  • Ion channels control the flow of ions through the cell membrane to determine the makjeonap and has a significant effect on cell function, such as to adjust the concentration of Ca 2 + in the excitable cells through it.
  • Ion channels are classified into Na + channels, Ca 2 + channels, and K + channels according to the type of ions to pass through.
  • K + channels that are activated by intracellular Ca 2 + There are several types of endothelial cells has KCa1.1 channel, KCa2.3 channel, KCa3.1 channel. These three channels play different roles in vascular endothelial cells.
  • KCa3.1 channels induce vascular endothelial hyperpolarization to regulate vasoconstriction (Tharp DL, Bowles DK, 2009, Cardiovasc Hematol Agents Med Chem. 7, 1- 11) plays an important role in new angiogenesis. In cells other than vascular endothelial cells, these channels play an important role in cell function regulation. KCa3.1 channels are also distributed in erythrocytes, fibroblasts, proliferating vascular smooth muscle cells, and immune cells (T cells and B cells) and contribute to the regulation of their function.
  • vascular endothelial cells play a very important role in the regulation of vascular aperture and contractility.
  • Vascular endothelial cells secrete NO or induce endothelial cell-dependent hyperpolarization to relax vascular smooth muscle and regulate vasoconstriction.
  • KCa3.1 channel When KCa3.1 channel is activated by intracellular calcium endothelial cell hyperpolarization is, vascular endothelial cell hyperpolarization promotes NO generation to induce intracellular Ca 2 + influx. Therefore, KCa3.1 channel regulates vascular smooth muscle contractility by inducing NO secretion as well as vascular endothelial cell hyperpolarization.
  • vascular diseases such as hypertension
  • relaxation due to hyperpolarization compensates for this and inhibits increase in vascular smooth muscle contractility.
  • KCa3.1 channels which play an important role in the function of vascular endothelial cells, are known to cause vascular diseases such as hypertension if they are not functioning properly.
  • KCa3.1 channel deficient mice KCa3.1 channel knockout mice
  • Hypertension may occur and decreased expression of KCa3.1 channels is known to cause diseases such as gestational hypertension.
  • vascular endothelial dysfunction is caused by vascular disease-causing substances in plasma.
  • vascular disease causing agents include oxidized low density-lipoprotein (oxLDL), hyperglycemia, and ROS such as superoxide generated by these substances.
  • oxLDL oxidized low density-lipoprotein
  • ROS ROS
  • vascular endothelial dysfunction occurs due to oxidized low density lipid protein in gestational hypertension and atherosclerosis, and hyperglycemia in diabetic vascular disease.
  • an increase in vascular contractility and a decrease in vascular caliber are caused, and blood flow is disturbed by an increase in vascular resistance, and gas and nutrient supply to tissues are not performed smoothly, causing vascular diseases.
  • These vascular disease-causing agents are known to cause vascular endothelial dysfunction by reducing the expression of vascular endothelial KCa3.1 channels (Choi S et al, 2013a, Free Radic Biol Med, 57, 10-21).
  • vascular endothelial dysfunction occurs when structural changes in blood vessels, ie, vascular diameter reduction, are caused by vascular endothelial dysfunction, and thus vascular endothelial dysfunction occurs before structural changes in blood vessels. Therefore, the diagnosis of vascular endothelial dysfunction can be used for early diagnosis of vascular disease (diagnosis of the disease before structural changes in blood vessels), progress and prognosis, and diagnosis of treatment effect.
  • gestational hypertension various methods such as measuring risk factors (age, etc.), uterine arterial blood flow using Doppler, and measuring gestational hypertension (sFlt-1, endoglin) in plasma are used for early diagnosis. No method has been developed to accurately diagnose the possibility of the disease. Accordingly, in the case of arteriosclerosis and diabetes, the progress of vascular disease is diagnosed by measuring blood flow and vascular diameter by angiography, CT angiography, Dopper Sono, etc. There is a problem that diagnosis is possible only after a structural change of the blood vessel (vascular change) is induced.
  • vascular change vascular change
  • the inventors of the present invention have made efforts to develop a method for more effectively diagnosing vascular endothelial dysfunction in order to more effectively manage and treat patients with vascular diseases.
  • the inventors of vascular endothelial cells or subjects treated with plasma or serum of subjects We can confirm that vascular endothelial dysfunction can be diagnosed by measuring the expression levels of Rab5C, clathrin, caveolin-1, Early Endosome Antigen 1 (EAE1) and KCa3.1 channels in erythrocytes.
  • EAE1 Early Endosome Antigen 1
  • One object of the present invention is to provide a composition for diagnosing malfunction of vascular endothelial cells.
  • Another object of the present invention to provide a kit for diagnosing dysfunction of vascular endothelial cells comprising the composition.
  • Still another object of the present invention is to provide a method for diagnosing vascular endothelial dysfunction.
  • Still another object of the present invention is to provide a method for screening a substance for improving the function of vascular endothelial cells.
  • Still another object of the present invention is to provide a method for screening a substance for inducing dysfunction of vascular endothelial cells.
  • the function of vascular endothelial cells using the expression levels of marker proteins such as Rab5C, clathrin, caveolin-1, and early endosome antigen 1 (EEA1). Since it is possible to diagnose whether there is a failure, it may be widely used for early diagnosis of various vascular diseases caused by dysfunction of vascular endothelial cells.
  • marker proteins such as Rab5C, clathrin, caveolin-1, and early endosome antigen 1 (EEA1). Since it is possible to diagnose whether there is a failure, it may be widely used for early diagnosis of various vascular diseases caused by dysfunction of vascular endothelial cells.
  • Figure 1 shows the results of comparing the expression level of KCa3.1 channel protein in umbilical vascular endothelial cells of normal mothers and gestational hypertension patients. ** means P ⁇ 0.01 compared to control (normal maternal tissue or vascular endothelial cells) (NP: normal mother, PE: gestational hypertension, EC: vascular endothelial cells).
  • Figure 1a is a micrograph and graph comparing the expression level of KCa3.1 channel protein by immunovascular staining in umbilical vein (left) and umbilical artery (right) of normal mother and gestational hypertension.
  • FIG. 1B is a photograph and a graph comparing or measuring the expression level of KCa3.1 channel protein by Western blot in umbilical vein tissue of normal mother and gestational hypertension patients.
  • Figure 1c is a photograph and a graph comparing or measuring the expression level of KCa3.1 channel protein in cultured umbilical vein endothelial cells by Western blot.
  • Figure 1d is a graph showing the results of measuring the KCa3.1 current in vascular endothelial cells cultured in the umbilical vein of normal mother and gestational hypertension patients.
  • Figure 2 shows the results of analyzing the effect of plasma or serum on the expression level of KCa3.1 channel protein of gestational hypertension patients (NP: normal mother, PE: gestational hypertension patients).
  • Figure 2a is a photograph and a graph showing the effect of gestational plasma or serum on the expression level of vascular endothelial KCa3.1 channel and the effect of antioxidant on it.
  • Figure 2b is a photograph showing the result of comparing the protein expression level of NOX2 (NADPH oxidase 2) in the plasma or serum of normal mothers and gestational hypertension patients.
  • NOX2 NADPH oxidase 2
  • FIG. 2C is a photograph and graph showing the effect of anti-LOX1 antibody on increasing protein expression level of NOX2 and decreasing protein expression level of KCa3.1 channel by gestational hypertension patients plasma or serum (LOX1: lectin-lik ox-LDL receptor-1, tempol and tiron: antioxidants).
  • ** means P ⁇ 0.01 compared to the control group (normal maternal plasma or serum-treated vascular endothelial cells), and # is a group treated with plasma or serum of gestational hypertension patients without LOX1 antibody.
  • Figure 3a is a photograph and graph showing the results of analyzing the effect of oxidized low density lipid protein (oxidized low density-lipoprotein, oxLDL) on vascular endothelial cell KCa3.1 channel protein expression level.
  • * and ** mean P ⁇ 0.05 and P ⁇ 0.01, respectively, compared to the control group (vascular endothelial cells not treated with vascular disease causing substances).
  • Figure 3b is a photograph and graph showing the results of analyzing the effect of hyperglycemia, a vascular disease causing substance, on the expression level of KCa3.1 channel protein of vascular endothelial cells.
  • * and ** mean P ⁇ 0.05 and P ⁇ 0.01, respectively, compared to the control group (vascular endothelial cells not treated with vascular disease causing substances).
  • Figure 4 is a graph and photograph showing the results of analyzing the effect of ROS on the expression level of vascular endothelial KCa3.1 channel protein.
  • 4 shows the effect of superoxide donor xantine and xantine oxidase mixture (Xantine and Xantine oxidase mixture, X / X0), and antioxidant (tempol, tiron) on the expression level of KCa3.1 channel protein .
  • ** is P ⁇ 0.01 compared to the control (vascular endothelial cells not treated with X / XO or antioxidant), and # and ## are blood vessels treated with X / XO (100 uM / 100 mU / ml).
  • P ⁇ 0.05 and P ⁇ 0.01 respectively.
  • NP normal mother
  • PE gestational hypertension patients
  • 5a is a photograph and a graph showing the effect of LPC, a vascular disease causing agent, on the protein expression level of vascular endothelial caveolin-1 and the effects of antioxidants on it.
  • Figure 5b is a photograph showing the effect of plasma or serum of gestational hypertension patients on the expression level of vascular endothelial caveolin-1 protein.
  • FIG. 5C is a photograph showing that the reduction of vascular endothelial KCa3.1 channel expression level by plasma or serum of gestational hypertension patients is induced by Cabolin-1.
  • ** means P ⁇ 0.01 compared to the control.
  • Figure 6 shows the results of analyzing the expression levels of clathrin, EEA1 (Early Endosome Antigen 1) according to the treatment of vascular disease causing agent.
  • Figure 6a is a photograph showing the results of observing the protein expression level of KCa3.1 channel, clathrin in the cell membrane of vascular endothelial cells treated with gestational hypertension patients plasma or serum.
  • Figure 6b is a photograph showing the results of observing the protein expression level of KCa3.1 channel, clathrin in the cell membrane of vascular endothelial cells treated with LPC.
  • Figure 6c is a photograph and a graph showing the result of comparing the expression level of EEA1 and KCa3.1 channel protein according to the concentration-specific treatment of X / X0.
  • Figure 7a is a photograph showing the results of comparing the expression level of Rab5C protein in vascular endothelial cells exposed to 10%, 25%, 100% concentration of gestational hypertension patients plasma or serum.
  • Figure 7b is a photograph showing the result of comparing the expression level of KCa3.1 channel protein in vascular endothelial cells exposed by diluting the plasma or serum of gestational hypertension patients to 10%, 25%, 100% concentration.
  • Figure 7c is a photograph showing the result of comparing the expression level of EEA1 protein in vascular endothelial cells exposed by diluting the plasma or serum of gestational hypertension patients to 10%, 25%, 100% concentration.
  • NP normal mother
  • PE gestational hypertension
  • FIG. 9 is a photograph and graph showing the results of analyzing the effect of TBHP treatment on the expression level of KCa3.1 channel protein in vascular endothelial cells.
  • one embodiment of the present invention provides a composition for diagnosing dysfunction of vascular endothelial cells.
  • the vascular endothelial dysfunction composition is composed of a group consisting of KCa3.1 channel and caveolin-1, Rab5C, clathrin, and Early Endosome Antigen 1 (EEA1). It may be a composition for diagnosing dysfunction of vascular endothelial cells, including an agent for measuring the level of mRNA of at least one protein or a gene thereof selected from the group.
  • vascular endothelial dysfunction causes vascular disease, but there is no direct way to measure vascular endothelial dysfunction. Therefore, by using the composition for diagnosing vascular endothelial dysfunction of the present invention, vascular endothelial dysfunction can be diagnosed early, the course, the prognosis, and the treatment effect. For example, in the case of gestational hypertension, using the composition of the present disease prior to the development of hypertension, the diagnosis of vascular endothelial dysfunction and the possibility of the development of gestational hypertension can be diagnosed, and thus new prophylaxis and treatment can be developed.
  • the diagnosis of vascular endothelial dysfunction using the composition of the present invention enables the diagnosis of vascular disease progression even before the symptoms of vascular disease appear, and thus the progress, prognosis, and treatment effect can be diagnosed Do.
  • caveolin-1 of the present invention is a major component of the caveolae plasma membrane found in most cell types, and is an early step in integrin binding in the Ras-ERK pathway. It is associated with phase and cell cycle progression.
  • the present inventors first identified the relationship between KCa3.1 channel and kaveolin-1 in vascular endothelial dysfunction.
  • the sequence information of the caveolin-1 can be obtained from a known database such as the National Center for Biotechnology Information (NCBI).
  • NCBI National Center for Biotechnology Information
  • Cabolin-1 of the present invention is NCBI GenBank Acession NO. NM_001172897.1, NM_001243064.1, NM_031556.3 or NM_001135818.1, but is not limited thereto.
  • Rab5C refers to a protein that plays a role in regulating membrane traffic by controlling fusion between early endosomes and plasma membranes as one of GTPases. .
  • the present inventors first identified the relationship with Rab5C in vascular endothelial dysfunction caused by a vascular disease causing agent.
  • Rab5C sequence information can be obtained from a known database such as the National Center for Biotechnology Information (NCBI).
  • NCBI National Center for Biotechnology Information
  • Rab5C of the present invention may be, but is not limited to, NCBI GenBank Acession NO.CR541901.1, AB232595.1, NM_001105840.2 or NM_001246383.1.
  • clathrin refers to a protein that exhibits a triskelion form having three branches composed of three heavy chains and three light chains. The three branches can interact to form a polyhedral lattice surrounding the vesicle. The present inventors first identified the relationship with clathrin in vascular endothelial dysfunction.
  • the sequence information of the clathrin can be obtained from a known database such as the National Center for Biotechnology Information (NCBI).
  • NCBI National Center for Biotechnology Information
  • the clathrin of the present invention may be prepared by NCBI GenBank Acession NO. NM_001288653.1, NM_001003908.1, NM_019299.1 or XM_001136053.4, but is not limited thereto.
  • Early Endosome Antigen 1 (EEA1) of the present invention is located in the early endosomes and plays an important role in endosomal trafficking.
  • the present inventors first identified the relationship between EEA1 in vascular endothelial dysfunction.
  • EEA1 can be obtained from a known database such as the National Center for Biotechnology Information (NCBI).
  • NCBI National Center for Biotechnology Information
  • EEA1 of the present invention is NCBI GenBank Acession NO. NM_003566.3, NM_001001932.3, NM_001108086.1 or XM_522610.5, but is not limited thereto.
  • vascular endothelial cells plasma or serum of gestational hypertension patients, oxLDL, LPC, hyperglycemia, etc.
  • protein expression levels of Cabolin-1, Clathrin, Rab5C and EEA1 As a result, it was confirmed that the expression level of KCa3.1 channel protein was decreased, whereas the protein expression levels of carveolin-1, clathrin, Rab5C and EEA1 were increased. This suggests that vascular endothelial dysfunction induced by vascular disease causing agents can be diagnosed by measuring the expression level of KCa3.1 channel, caveolin-1, clathrin, Rab5C or EEA1.
  • the expression level of Rab5C and EEA1 protein increased, about 1/10 It was confirmed that Rab5C and EEA1 protein expression levels were increased only by gestational hypertension inducing factor alone. In such a small amount, the expression level of the marker can be measured to diagnose vascular endothelial dysfunction, thereby predicting or early diagnosis of the possibility of vascular disease.
  • the subject means all animals including humans, and specifically, may be human, but is not limited thereto.
  • vascular endothelial cell refers to one of the major cells that form the structure of the blood vessels, mainly serves to maintain vascular normality, such as expansion and contraction of blood vessels, proliferation and migration of vascular smooth muscle, thrombogenesis and lysis do.
  • the dysfunction of the vascular endothelial cells causes vascular wall damage and increased vascular contractility, through which vascular diseases occur.
  • the vascular disease caused by the dysfunction of the vascular endothelial cells is not particularly limited thereto.
  • the vascular disease may be hypertension, arteriosclerosis, diabetic vascular disease, or a combination thereof. Can be.
  • the vascular disease is caused by the dysfunction of vascular endothelial cells
  • the progress of vascular disease can be known according to the level of vascular endothelial dysfunction, and the vascular endothelial cells Dysfunction may also be caused.
  • dysfunction of the vascular endothelial cells can change the KCa3.1 channel expression level of the vascular endothelial cells
  • KCa3.1 channel expression level of the vascular endothelial cells may be affected by the substance causing the vascular disease. It may be.
  • the vascular disease-causing substance can cause vascular endothelial dysfunction, cause the development of vascular disease, or change KCa3.1 channel expression level of vascular endothelial cells,
  • it can be, for example, a substance that increases oxidative stress, and in another example, oxidized low-density lipoprotein, lysophosphatidylcholine (LPC), which can increase oxidative stress, Gestational hypertension (sFlt-1, endoglin), high blood sugar, superoxide anion donor (superoxide anion donor).
  • diagnosis refers to a series of acts of identifying the presence or characteristic of a pathological condition.
  • the diagnosis may be interpreted to mean the act of checking whether the vascular endothelial dysfunction is impaired, thereby predicting the onset of vascular disease due to the vascular endothelial dysfunction.
  • composition for diagnosing dysfunction of vascular endothelial cells of the present invention may further include an agent for measuring KCa3.1 channel protein or its mRNA level.
  • KCa3.1 channel (intermediate conductance calcium-activated potassium channel, subfamily N, member 4) refers to a voltage-independent potassium channel in the form of a heterotetramer that is activated by intracellular calcium.
  • KCa3.1 channels of vascular endothelial cells induce hyperpolarization to relax blood vessels, while hyperpolarization increases intracellular Ca2 + influx and promotes NO production by eNOS, thereby relaxing blood vessels.
  • eNOS intracellular Ca2 + influx and promotes NO production by eNOS, thereby relaxing blood vessels.
  • vascular endothelial dysfunction may be induced.
  • By analyzing the expression level of the KCa3.1 channel protein or gene it is possible to determine whether the vascular endothelial dysfunction is induced.
  • the expression level of KCa3.1 channel protein decreases in gestational hypertension patients. Confirmed.
  • oxLDL, LPC and hyperglycemia as a result of analyzing the effect of plasma or serum, oxLDL, LPC and hyperglycemia on vascular endothelial cells of normal maternal and gestational hypertension patients gestational hypertension patients, gestational It was confirmed that the expression level of KCa3.1 channel protein is reduced by plasma or serum, oxLDL, LPC and hyperglycemia of hypertensive patients. As a result, it was found that oxLDL and hyperglycemia, which are plasma or serum substances, abnormally reduce KCa3.1 channel expression of vascular endothelial cells in gestational hypertension and vascular complications of diabetes, thereby inducing vascular endothelial dysfunction.
  • oxidative stress on the KCa3.1 channel protein expression level in vascular endothelial cells of normal mothers and gestational hypertension patients
  • KCa3.1 channel protein expression through oxidant treatment It was confirmed that the decrease, and recovered by the antioxidant. Through this, oxidative stress caused vascular endothelial dysfunction, and it was found that oxidative stress level can be measured by measuring the expression level of KCa3.1 channel protein.
  • KCa3.1 channel protein expression level was measured in normal vascular endothelial cells (HUVECs) exposed to plasma or serum of gestational hypertension patients diluted in various stages. Since dilution to about 1/10 diminishes the expression level, it was confirmed that measuring the expression level of the KCa3.1 channel can predict the onset or early diagnosis of vascular disease.
  • mRNA level measurement is a method of confirming the expression level of genes for diagnosing vascular endothelial dysfunction in isolated cells for diagnosing vascular endothelial dysfunction. Means the way. Specific methods for measuring the mRNA level is not particularly limited, but as an example, reverse transcriptase (RT-PCR), competitive reverse transcriptase (Competitive RT-PCR), real-time reverse transcriptase (Real- time RT-PCR), RNase protection assay (RPA), Northern blotting, DNA chips and the like can be used.
  • RT-PCR reverse transcriptase
  • Competitive RT-PCR competitive reverse transcriptase
  • Real- time RT-PCR real-time reverse transcriptase
  • RNase protection assay RNase protection assay
  • the agent used to measure the mRNA level is specifically a primer pair, a probe or an antisense oligonucleotide, and since the nucleic acid information of the genes is known to GeneBank, etc., those skilled in the art can specifically detect specific regions of these genes based on the sequence.
  • Primer pairs, probes or antisense oligonucleotides can be designed.
  • the agent for measuring the mRNA level is a primer pair, a probe specifically binding to a gene encoding a protein selected from Rab5C, clathrin, caveolin-1 and EEA1 Or antisense oligonucleotides.
  • primer pair of the present invention is not particularly limited as long as it can recognize and amplify a gene sequence encoding KCa3.1 channel, Rab5C, clathrin, caveolin-1 or EEA1 provided herein.
  • it may be a primer pair of any combination consisting of forward and reverse primers capable of binding to each gene, and as another example, may be a primer pair capable of binding with specificity and sensitivity to each gene.
  • the primer consists of a sequence whose nucleic acid sequence is inconsistent with the non-target sequence present in the sample, thus amplifying only the target gene sequence containing the complementary primer binding site and not causing nonspecific amplification. It may be a primer pair having.
  • probe refers to a substance that can specifically bind to a target substance to be detected in a sample, and means a substance that can specifically confirm the presence of the target substance in the sample through the binding.
  • Component of the probe is not particularly limited as long as it can specifically bind to the target material, for example, peptide nucleic acid (PNA), locked nucleic acid (LNA), peptides, polypeptides, proteins, RNA, DNA and the like, and as other examples, DNA such as cDNA, genomic DNA, etc .; RNA such as genomic RNA, mRNA and the like; And proteins such as antibodies, antigens, enzymes, peptides, and the like.
  • PNA peptide nucleic acid
  • LNA locked nucleic acid
  • antisense oligonucleotide refers to a DNA or RNA or derivative thereof containing a nucleic acid sequence complementary to a sequence of a particular mRNA, which binds to a complementary sequence in the mRNA and inhibits translation of the mRNA into a protein. Do it.
  • Antisense oligonucleotide sequence refers to a DNA or RNA sequence that is complementary to the mRNA of said genes and capable of binding to said mRNA. This may inhibit the essential activity of the translation of the gene mRNA, translocation into the cytoplasm, maturation or any other overall biological function.
  • the length of the antisense oligonucleotide may be 6 to 100 bases, specifically 8 to 60 bases, more specifically 10 to 40 bases.
  • the antisense oligonucleotides may be synthesized in vitro in a conventional manner to be administered in vivo or to allow the antisense oligonucleotides to be synthesized in vivo.
  • One example of synthesizing antisense oligonucleotides in vitro is using RNA polymerase I.
  • One example of allowing antisense RNA to be synthesized in vivo is to allow the antisense RNA to be transcribed using a vector whose origin is in the opposite direction of the multicloning site (MCS).
  • MCS multicloning site
  • the antisense RNA is preferably such that the translation stop codon is present in the sequence so that it is not translated into the peptide sequence.
  • the term "measurement of protein levels" of the present invention refers to the KCa3.1 channel, Rab5C, clathrin, caveolin-1, or EEA1, a marker protein for diagnosing vascular endothelial dysfunction in isolated cells for diagnosing vascular endothelial dysfunction. It means the process of checking the presence and the degree of expression.
  • the method for measuring the protein level is as long as it can measure the expression level of the marker protein KCa3.1 channel, Rab5C, clathrin, caveolin-1 or EEA1, which is a diagnostic marker protein for vascular endothelial dysfunction provided by the present invention.
  • examples include, but are not limited to, protein chip analysis, immunoassay, ligand binding assay, Matrix Desorption / Ionization Time of Flight Mass Spectrometry (MALDI-TOF) analysis, and Surface Enhanced Laser Desorption / Ionization Time of Flight (SELDI-TOF).
  • Mass Spectrometry Analysis Radioimmunoassay, Radioimmunoassay, Oukteroni Immunodiffusion, Rocket Immunoelectrophoresis, Tissue Immunostaining, Complementary Assay, Two-Dimensional Electrophoresis, Liquid Chromatography-Mass Spectrometry, LC-MS), liquid chromatography-mass spectrometry / mass spectrometry, western blot, and enzyme linked immunosorbentassay (ELISA).
  • the method for measuring the protein level, KCa3.1 channel, Rab5C, clathrin, caveolin-1 or a marker protein for diagnosing vascular endothelial dysfunction to diagnose vascular endothelial dysfunction It can be interpreted by using an antibody that specifically binds to EEA1.
  • antibody refers to a specific protein molecule directed against an antigenic site.
  • the antibody may be interpreted to mean an antibody that specifically binds to a KCa3.1 channel, Rab5C, clathrin, caveolin-1, or EEA1 protein, which is a marker protein for diagnosing vascular endothelial dysfunction. .
  • the antibody may include all polyclonal antibodies, monoclonal antibodies, recombinant antibodies and the like, and may include functional fragments of antibody molecules as well as complete forms having two full length light chains and two full length heavy chains. have.
  • the functional fragment of the antibody molecule means a fragment having at least antigen binding function, and examples thereof include Fab, F (ab '), F (ab') 2, and Fv.
  • the present invention provides a kit for diagnosing dysfunction of vascular endothelial cells, comprising the composition for diagnosing dysfunction of vascular endothelial cells.
  • the kit for diagnosing dysfunction of vascular endothelial cells is not particularly limited thereto.
  • a reverse transcription polymerase chain reaction (RT-PCR) kit a DNA chip kit, an enzyme-linked immunosorbent assay (ELISA) kit, Protein chip kits, rapid kits, multiple reaction monitoring (MRM) kits, and the like.
  • RT-PCR reverse transcription polymerase chain reaction
  • DNA chip kit a DNA chip kit
  • ELISA enzyme-linked immunosorbent assay
  • Protein chip kits Protein chip kits
  • rapid kits multiple reaction monitoring (MRM) kits, and the like.
  • MRM multiple reaction monitoring
  • the kit for diagnosing dysfunction of vascular endothelial cells may further include one or more other component compositions, solutions or devices suitable for analytical methods.
  • the kit may be a diagnostic kit comprising essential elements necessary to perform reverse transcriptase.
  • the reverse transcription polymerase kit contains each primer pair specific for the marker gene.
  • a primer is a nucleotide having a sequence specific to the nucleic acid sequence of each gene, the length of about 7 bp to 50 bp, more specifically about 10 bp to 30 bp in length. It may also include primers specific for the nucleic acid sequence of the control gene.
  • reverse transcriptase kits include test tubes or other suitable containers, reaction buffers (pH and magnesium concentrations vary), enzymes such as deoxynucleotides (dNTPs), Taq-polymerase and reverse transcriptase, DNAse, RNAse inhibitor DEPC- It may include DEPC-water, sterile water, and the like.
  • reaction buffers pH and magnesium concentrations vary
  • enzymes such as deoxynucleotides (dNTPs), Taq-polymerase and reverse transcriptase
  • DNAse DNAse
  • RNAse inhibitor DEPC- It may include DEPC-water, sterile water, and the like.
  • the kit may be a diagnostic kit, which includes an essential element necessary to perform a DNA chip.
  • the DNA chip kit may include a substrate on which a cDNA or oligonucleotide corresponding to a gene or a fragment thereof is attached, and a reagent, a preparation, an enzyme, or the like for preparing a fluorescent probe.
  • the substrate may also comprise cDNA or oligonucleotide corresponding to the control gene or fragment thereof.
  • the kit may be a diagnostic kit characterized by including essential elements necessary to perform an ELISA.
  • ELISA kits include antibodies specific for the KCa3.1 channel, Rab5C, clathrin, caveolin-1 and EEA1 proteins.
  • Antibodies are antibodies that have high specificity and affinity for each marker protein and have little cross-reactivity to other proteins. They are monoclonal antibodies, polyclonal antibodies, or recombinant antibodies.
  • the ELISA kit can also include antibodies specific for the control protein.
  • Other ELISA kits can bind reagents that can detect bound antibodies, such as labeled secondary antibodies, chromophores, enzymes (eg conjugated with the antibody) and substrates or antibodies thereof. Other materials and the like.
  • the kit may be a rapid kit, characterized in that it contains the essential elements necessary to perform a rapid test that can be analyzed within 5 minutes.
  • Rapid kits include antibodies specific for a protein. Antibodies are antibodies that have high specificity and affinity for each marker protein and have little cross-reactivity to other proteins. They are monoclonal antibodies, polyclonal antibodies, or recombinant antibodies. Rapid kits may also include antibodies specific for the control protein. Other rapid kits include reagents that can detect bound antibodies, such as nitrocellulose membranes to which specific and secondary antibodies are immobilized, membranes bound to beads to which antibodies are bound, absorbent pads, and sample pads. Substances and the like.
  • the kit may be a multiple reaction monitoring (MRM) kit, which is an MS / MS mode characterized by including the necessary elements necessary to perform mass spectrometry.
  • MRM multiple reaction monitoring
  • SIM Selected Ion Monitoring
  • MRM selects a particular ion one more time from the broken ion to detect another continuously connected source of MS. It is a method of using the ions obtained from the collision after passing through it once more. More specifically, in SIM, there is a problem in that the selected ions may interfere with quantification when the selected ions are ions that are also detected in plasma.
  • MRM multiple reaction monitoring
  • the present invention provides a method for diagnosing malfunction of vascular endothelial cells.
  • the Rab5C, clathrin, caveolin-1, EEA1 and KCa3.1 channels are characterized by a change in the expression level of the gene or the level of the protein in an individual with dysfunction of vascular endothelial cells. If the vascular endothelial dysfunction can be diagnosed, it can be diagnosed early on the development of vascular disease or the progress of the vascular disease.
  • the method for diagnosing vascular endothelial dysfunction of the present invention includes (a) Rab5C, clathrin, caveolin-1, EEA1, and the like in biological samples of individuals suspected of vascular endothelial dysfunction. Measuring protein or mRNA levels of a marker selected from the group consisting of a combination of; And (b) comparing the measured level (i) with the level measured from a normal control sample.
  • the protein or mRNA level measurement is the same as defined above.
  • biological sample refers to a sample collected from the individual for diagnosing vascular endothelial dysfunction of a suspected vascular endothelial dysfunction, particularly as long as the vascular endothelial dysfunction can be confirmed.
  • the present invention may be, but is not limited to, whole blood, plasma, serum, fractions thereof, or cells contained therein, and red blood cells.
  • the method may further comprise the step (a) measuring the protein or mRNA level of (ii) KCa3.1 channel.
  • the method measures the protein or mRNA level of (ii) KCa3.1 channel in step (a), and using the measured (i) and (ii) calculated by the formula of (i) / (ii) Comparing the calculated rate to the rate calculated from the normal control sample.
  • Rab5C, clathrin, caveolin-1 or EEA1 protein or mRNA levels in biological samples of individuals suspected of vascular endothelial dysfunction are relatively higher than those measured from normal control samples.
  • KCa3.1 channel protein As a result of confirming the expression level of KCa3.1 channel and Rab5C protein in red blood cells, it was confirmed that the expression level of KCa3.1 channel protein is decreased and the expression level of Rab5C protein is increased. Through this, the expression levels of KCa3.1 channel protein and Rab5C protein were measured in erythrocytes, indicating that vascular endothelial dysfunction could be diagnosed.
  • the method for diagnosing dysfunction of vascular endothelial cells of the present invention may comprise the steps of: (a) treating a biological sample of a subject suspected of dysfunction of vascular endothelial cells to normal vascular endothelial cells; (b) protein or mRNA levels of markers selected from the group consisting of (i) Rab5C, clathrin, caveolin-1, EEA1, and combinations thereof in vascular endothelial cells treated with the biological sample of step (a) Measuring; And (c) comparing said measured level (i) with a level measured from normal vascular endothelial cells to which said biological sample has not been treated.
  • the protein or mRNA level measurement is the same as defined above.
  • the method may further comprise the step of (ii) (ii) measuring the protein or mRNA level of the KCa3.1 channel.
  • the method measures the protein or mRNA level of (ii) KCa3.1 channel in step (b), and using the measured (i) and (ii) calculated by the formula of (i) / (ii) Comparing the calculated rate to a rate calculated from normal vascular endothelial cells to which the biological sample has not been treated.
  • the protein or mRNA level of Rab5C, clathrin, caveolin-1, or EEA1 measured in vascular endothelial cells treated with a biological sample of an individual suspected of vascular endothelial dysfunction is the biological The sample exhibited a relatively higher level than that measured from untreated normal vascular endothelial cells, or (ii) the protein or mRNA levels of the KCa3.1 channel were measured from normal vascular endothelial cells not treated with the biological sample. If the level is relatively low, it may be determined that dysfunction of vascular endothelial cells has occurred in the subject.
  • the present invention provides a method for screening a substance for inducing a function of vascular endothelial cells.
  • a candidate substance predicted to induce improvement of vascular endothelial cells is treated to vascular endothelial cells in which dysfunction has occurred, and a failure marker of vascular endothelial cells expressed in the vascular endothelial cells to which the candidate substance has been treated.
  • the candidate substance is endothelium It can be determined whether it is possible to induce improvement of the function of the cell.
  • the method for screening a function improving inducer of vascular endothelial cells of the present invention comprises the steps of: (a) treating candidate cells expected to be capable of inducing functional improvement of vascular endothelial cells to separated cells; (b) measuring the protein or mRNA levels of a marker selected from the group consisting of (i) Rab5C, clathrin, caveolin-1, EEA1, and combinations thereof in cells treated with the candidate substance; And (c) comparing the level of the marker measured in step (b) with the level of the marker measured in isolated cells not treated with the candidate substance.
  • the candidate material refers to a substance capable of regulating dysfunction of vascular endothelial cells, for example, to restore vascular endothelial cells in which dysfunction has occurred to a normal state, or It may be a substance that is expected to exhibit or induce an effect that can alleviate, ameliorate, or treat dysfunction, and as another example, may cause dysfunction from vascular endothelial cells or prevent dysfunction of vascular endothelial cells. It may be a substance that is expected to be inducible.
  • the candidate is not particularly limited as long as it is expected to have an effect of regulating dysfunction of vascular endothelial cells, but as an example, any substance, molecule, element, compound ), Entities, etc., and may be other examples, such as natural products, synthetic compounds, and the like, and as another example, proteins, polypeptides, small organic molecules, polysaccharides, and the like. (polysaccharide), polynucleotide and the like.
  • the isolated cell is not particularly limited as long as it can be used as a target for confirming whether or not it can induce improvement of vascular endothelial cell function of the candidate, but as an example, vascular endothelial cell function Cells isolated from the subject causing the failure, as another example, cells isolated from the blood or blood vessels of the subject causing vascular endothelial dysfunction, and as another example, vascular endothelial dysfunction Erythrocytes isolated from the blood of the induced individual or vascular endothelial cells isolated from the blood vessels of the individual.
  • the method may further comprise the step of (ii) (ii) measuring the protein or mRNA level of the KCa3.1 channel.
  • the method measures the protein or mRNA level of (ii) KCa3.1 channel in step (b), and using the measured (i) and (ii) calculated by the formula of (i) / (ii) Comparing the proportions calculated to the proportions calculated from cells in which the candidate material has not been treated.
  • (i) protein or mRNA levels of Rab5C, clathrin, caveolin-1 or EEA1 measured in cells treated with the candidate are relatively higher than those measured from cells not treated with the candidate.
  • the candidate is a function of vascular endothelial cells if the protein or mRNA level of the KCa3.1 channel is at a lower level than that measured from cells not treated with the candidate. It can be determined that improvement can be induced.
  • the candidate material is vascular endothelial. It can be determined that the improvement of the function of the cell can be induced.
  • KCa3.1 channel protein when treated with TBHP, a vascular endothelial function inducing substance, the expression level of KCa3.1 channel protein was confirmed to increase significantly in concentration. It is known that an increase in KCa3.1 channel expression levels can improve vascular endothelial function (Chadha PS et al, 2010, J Pharmacol Exp Ther, 335 (2), 284-293). After treating candidate substances that are expected to induce improvement, the protein expression levels of KCa3.1 channels are measured, suggesting that the candidates can determine whether they can induce improvement of vascular endothelial function. will be.
  • the present invention provides a method for screening a substance for inducing dysfunction of vascular endothelial cells.
  • vascular endothelial dysfunction occurs, the expression levels of the Rab5C, clathrin, caveolin-1, EEA1 and KCa3.1 channels are changed, which may induce improvement of vascular endothelial cells.
  • Candidates predicted to be treated are treated with vascular endothelial cells, and the candidate substances are treated with Rab5C, clathrin, caveolin-1, EEA1, and KCa3.1 channels, which are dysfunction markers of vascular endothelial cells. After measuring the expression level, it is possible to determine whether the candidate substance can induce dysfunction of vascular endothelial cells by checking whether the expression level of each measured marker is changed.
  • the method of screening a substance for inducing dysfunction of vascular endothelial cells of the present invention comprises the steps of: (a) treating a candidate material that is expected to induce dysfunction of vascular endothelial cells to an isolated cell; (b) measuring the protein or mRNA levels of a marker selected from the group consisting of (i) Rab5C, clathrin, caveolin-1, EEA1, and combinations thereof in cells treated with the candidate substance; And (c) comparing the level of the marker measured in step (b) with the level of the marker measured in isolated cells not treated with the candidate substance.
  • the candidates and the isolated cells are the same as defined above.
  • the method may further comprise the step of (ii) (ii) measuring the protein or mRNA level of the KCa3.1 channel.
  • the method measures the protein or mRNA level of (ii) KCa3.1 channel in step (b), and using the measured (i) and (ii) calculated by the formula of (i) / (ii) Comparing the proportions calculated to the proportions calculated from cells in which the candidate material has not been treated.
  • (i) protein or mRNA levels of Rab5C, clathrin, caveolin-1 or EEA1 measured in cells treated with the candidate are relatively higher than those measured from cells not treated with the candidate.
  • the candidate material functions as a vascular endothelial cell if the protein or mRNA level of the KCa3.1 channel is relatively lower than that measured from cells not treated with the candidate material. It can be determined that failure can be induced.
  • the candidate material when the ratio of (i) / (ii) calculated in the cells treated with the candidate material is relatively higher than that calculated from the cells in which the candidate material is not treated, the candidate material is vascular endothelium. It can be determined that the cell can be inferior in function.
  • KCa3.1 channel protein when treated with vascular disease-causing LPC or X / XO to normal vascular endothelial cells, KCa3.1 channel protein is reduced and the expression level of the caveolin-1, clathrin, EEA1 protein is increased It was.
  • the expression level of the protein is measured to determine whether the candidate substance can induce vascular endothelial dysfunction. It can be seen that judging.
  • Example 1 Endothelial Cells in Patients with Vascular Disease KCa3 . 1 channel Protein expression level analysis
  • KCa3.1 channel expression decreases in vascular endothelial cells of patients with vascular disease
  • the expression levels of KCa3.1 channel proteins in vascular endothelial cells of normal mothers and gestational hypertension patients were compared.
  • KCa3.1 channel protein expression levels in umbilical veins and umbilical artery tissues were measured by histological analysis (A in FIG. 1), Western blot (B in FIG. 1), and vascular endothelial cells in umbilical veins in normal maternal and gestational hypertension.
  • the isolation culture was performed to measure the expression level of KCa3.1 channel protein (C in FIG. 1) and the magnitude of KCa3.1 current (D in FIG. 1).
  • antibodies having the amino acid sequence of RQVRLKHRKLREQV SEQ ID NO: 1 were used.
  • Example 2 vascular endothelial cells caused by vascular disease causing substances KCa3 . 1 channel Protein expression level analysis
  • KCa3.1 channel expression in vascular endothelial cells of patients with vascular disease is caused by vascular disease causing agents in plasma or serum, plasma or serum, oxLDL, LPC and hyperglycemia of normal maternal and gestational hypertension patients
  • vascular disease causing agents in plasma or serum, plasma or serum, oxLDL, LPC and hyperglycemia of normal maternal and gestational hypertension patients The effect on the expression level of KCa3.1 channel in endothelial cells was analyzed.
  • the plasma or serum of gestational hypertension increased the expression level of NOX2 protein through LOX-1, increased ROS production and decreased KCa3.1 channel protein expression level, and major vascular disease of plasma or serum of gestational hypertension patients. It was found that the trigger was oxLDL. In addition, the expression level of KCa3.1 channel protein in vascular endothelial cells also decreased by oxLDL or hyperglycemia (FIG. 3).
  • oxLDL and hyperglycemia which are plasma or serum substances, abnormally reduce KCa3.1 channel expression levels of vascular endothelial cells in gestational hypertension and vascular complications of diabetes mellitus, thereby inducing vascular endothelial dysfunction.
  • Example 3 oxidative stress in vascular endothelial cells KCa 3.1 Expression level analysis of channels
  • KCa3.1 channel protein was measured using Western blot. In each sample, the expression level of ⁇ -tubulin or ⁇ -actin was measured as a control.
  • KCa3.1 channel protein expression level was decreased by oxidizing agent (X / XO) treatment, and KCa3.1 channel protein expression level was decreased by oxidizing agent (FIG. 4).
  • Example 4 vascular endothelial cells following treatment with vascular disease causing agent caveolin Analysis of Expression Levels of --1, clathrin, Rab-5C, and EEA1 Protein
  • caveolin-1, clathrin, EEA1 protein is increased by the vascular disease causing agent.
  • vascular endothelial dysfunction induced by vascular disease causing agent can be diagnosed by measuring the expression level of KCa3.1 channel or caveolin-1, clathrin, EEA1.
  • Example 5 Analysis of KCa3.1 channel, Rab5C, EEA1 protein expression levels in vascular endothelial cells according to the plasma or serum dilution of gestational hypertension
  • the serum was mixed with the culture medium, and the human vascular endothelial cells incubated in a solution containing plasma or serum concentrations of 10, 25, 100% (without dilution) were exposed for 24 hours, followed by Western blot using KCa3.
  • the expression level of Rab5C protein and the expression level of EEA1 protein were measured by the expression level of .1 channel protein, small GTPase. In each sample, the expression level of ⁇ -actin was measured as a control.
  • the expression level of Rab5C and EEA1 protein was increased by only about 1/10 of gestational hypertension-inducing factor and vascular endothelial cell KCa3.1 channel protein expression compared to the case where gestational hypertension was fully expressed. It was confirmed that it causes a decrease in levels.
  • Example 6 in red blood cells KCa3 . 1 channel Rab5C Protein expression level analysis
  • KCa3.1 channels Rab5C and clathrin in vascular endothelial cells and erythrocytes is the same in patients with vascular endothelial dysfunction
  • KCa3 Expression levels of one channel, Rab5C and clathrin protein were measured. In each sample, the expression level of ⁇ -actin was measured as a control.
  • vascular endothelial KCa3.1 channel protein in gestational hypertension patients can be determined by measuring KCa3.1 channel protein or Rab5C, clathrin, etc. in erythrocytes.
  • Example 7 TBHP Endothelial Cells Following Treatment KCa3 . 1 channel Expression level analysis of proteins
  • KCa3.1 channel expression levels can improve vascular endothelial function (Chadha PS et al, 2010, J Pharmacol Exp Ther, 335 (2), 284-293).
  • the protein expression level of KCa3.1 channel can be measured to determine whether the candidate substance can induce improvement of vascular endothelial function. Is to suggest.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Analytical Chemistry (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Zoology (AREA)
  • Physics & Mathematics (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Pathology (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • General Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

La présente invention concerne une composition, un kit, et un procédé de diagnostic d'un dysfonctionnement de cellules endothéliales vasculaires et un procédé de dépistage d'une substance qui cause le dysfonctionnement ou qui améliore la fonction des cellules endothéliales vasculaires. Le procédé décrit selon la présente invention permet, lorsqu'il est utilisé, le diagnostic d'un dysfonctionnement des cellules endothéliales vasculaires en utilisant les niveaux d'expression de protéines marqueurs, telles que la Rab5C, la clathrine, la cavéoline-1, et l'antigène endosomique précoce 1 (EEA1), et peut donc être largement employé pour le diagnostic précoce de diverses maladies vasculaires déclenchées par un dysfonctionnement des cellules endothéliales vasculaires.
PCT/KR2016/011223 2015-10-07 2016-10-07 Composition destinée au diagnostic du dysfonctionnement de cellules endothéliales vasculaires, et son utilisation Ceased WO2017061809A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020150141197A KR101889594B1 (ko) 2015-10-07 2015-10-07 혈관내피세포 기능 부전 진단용 조성물 및 이의 용도
KR10-2015-0141197 2015-10-07

Publications (1)

Publication Number Publication Date
WO2017061809A1 true WO2017061809A1 (fr) 2017-04-13

Family

ID=58488147

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2016/011223 Ceased WO2017061809A1 (fr) 2015-10-07 2016-10-07 Composition destinée au diagnostic du dysfonctionnement de cellules endothéliales vasculaires, et son utilisation

Country Status (2)

Country Link
KR (1) KR101889594B1 (fr)
WO (1) WO2017061809A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20240110699A (ko) * 2023-01-06 2024-07-16 셀라이온바이오메드 주식회사 KCa3.1 채널 단백질에 특이적으로 결합하는 단일클론 항체 및 이를 포함하는 혈관질환 진단용 조성물

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101517689B1 (ko) * 2013-10-07 2015-05-04 이화여자대학교 산학협력단 신규한 KCa3.1 채널 항체 및 이의 용도
KR20150085459A (ko) * 2014-01-14 2015-07-23 주식회사 엘지생명과학 대장암 마커로서의 신규 ntrk1 융합유전자 및 이의 용도
KR101552021B1 (ko) * 2013-11-22 2015-09-09 이화여자대학교 산학협력단 Rab5 발현 저해 물질을 유효 성분으로 포함하는 파브리 질환의 예방 또는 치료용 조성물

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101517689B1 (ko) * 2013-10-07 2015-05-04 이화여자대학교 산학협력단 신규한 KCa3.1 채널 항체 및 이의 용도
KR101552021B1 (ko) * 2013-11-22 2015-09-09 이화여자대학교 산학협력단 Rab5 발현 저해 물질을 유효 성분으로 포함하는 파브리 질환의 예방 또는 치료용 조성물
KR20150085459A (ko) * 2014-01-14 2015-07-23 주식회사 엘지생명과학 대장암 마커로서의 신규 ntrk1 융합유전자 및 이의 용도

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ACHCAR ET AL.: "Loss of Caveolin and Heme Oxygenase Expression in Severe Pulmonary Hypertension", CHEST, vol. 129, no. 3, March 2006 (2006-03-01), pages 696 - 705, XP055077271 *
DRAB ET AL.: "Loss of Caveolae, Vascular Dysfunction, and Pulmonary Defects in Caveolin-1 Gene -disrupted Mice", SCIENCE, vol. 293, 28 September 2001 (2001-09-28), pages 2449 - 2452, XP055377963 *
FERNANDEZ-HERNANDO ET AL.: "Endothelial-specific Overexpression of Caveolin-1 Accelerates Atherosclerosis in Apolipoprotein E-deficient Mice", THE AMERICAN JOURNAL OF PATHOLOGY, vol. 177, no. 2, August 2010 (2010-08-01), pages 998 - 1003, XP055377958 *
SUH, SUK-HYO ET AL.: "The Underlying Mechanisms of Vascular Endothelial and Smooth Muscle Dysfunctions in Preeclampsia Patient", MINISTRY OF HEALTH AND WELFARE, HEALTH & MEDICAL TECHNOLOGY R&D PROGRAM FINAL REPORT, RESEARCH MANAGING DEPARTMENT: EWHA UNIVERSITY - INDUSTRY COLLABORATION FOUNDATION, 18 June 2012 (2012-06-18) *

Also Published As

Publication number Publication date
KR101889594B1 (ko) 2018-08-21
KR20170041960A (ko) 2017-04-18

Similar Documents

Publication Publication Date Title
WO2019039817A2 (fr) Composition de prédiction ou de diagnostic d'hépatopathie et méthode de prédiction ou de diagnostic d'hépatopathie l'utilisant
WO2012081898A2 (fr) Marqueur destiné à établir un pronostic du cancer de l'estomac et procédé d'établissement d'un pronostic du cancer de l'estomac
WO2009113814A2 (fr) Marqueur protéique utilisé pour le diagnostic précoce du cancer du foie
WO2018038452A1 (fr) Biomarqueur sbp1 pour le diagnostic précoce de maladies rénales et utilisation associée
Yin et al. Pro-inflammatory Macrophages suppress PPARγ activity in Adipocytes via S-nitrosylation
WO2019182371A1 (fr) Composition pour le diagnostic de maladies neurologiques dégénératives
WO2018097646A1 (fr) Composition de diagnostic de maladies
WO2013187730A1 (fr) Composition pour la prévention ou le traitement d'une fracture ou de l'ostéoporose à l'aide d'un système slit-robo
WO2013009146A2 (fr) Marqueur pour le diagnostic d'une rétinopathie diabétique
WO2023234485A1 (fr) Biomarqueur pour prédire la probabilité de développer un cancer gastrique et son utilisation
WO2017061809A1 (fr) Composition destinée au diagnostic du dysfonctionnement de cellules endothéliales vasculaires, et son utilisation
Brunner et al. Validation of antibody-based tools for galanin research
WO2017018735A1 (fr) Biomarqueur de détermination du vieillissement, de détermination de l'obésité, et de diagnostic du cancer et trousse de diagnostic l'utilisant
WO2013100273A1 (fr) Marqueur diagnostique d'exposition à un rayonnement igfbp-5, composition permettant de diagnostiquer l'exposition à un rayonnement par mesure du niveau d'expression du marqueur, kit de diagnostic d'exposition à un rayonnement comprenant la composition, et méthode permettant de diagnostiquer l'exposition à un rayonnement faisant appel audit marqueur
WO2017082655A1 (fr) Procédés pour déterminer la résistance à une thérapie anticancéreuse et composition utilisée à cet effet
WO2019225923A1 (fr) Composition pour diagnostiquer des maladies
WO2013009143A2 (fr) Marqueur pour le diagnostic d'une rétinopathie diabétique
WO2020197227A1 (fr) Biomarqueur céréblon pour le diagnostic d'un carcinome hépatocellulaire, et nouvel anticorps monoclonal spécifique de celui-ci
WO2023090901A1 (fr) Procédé de sélection d'un marqueur de sénescence cellulaire utilisant un apprentissage automatique, biomarqueur pour la sénescence cellulaire, et procédé de criblage d'agent senolytique l'utilisant
WO2018088826A2 (fr) Composition de diagnostic de métastases du cancer colorectal ou de prédiction de pronostic et son utilisation
WO2023144708A1 (fr) Nouveau biomarqueur pour le diagnostic du cancer de l'ovaire
WO2023008858A1 (fr) Nouvel anticorps pour prévenir ou traiter une maladie fibrotique
WO2022145733A1 (fr) Prédiction de l'efficacité d'un agent thérapeutique ciblant un cancer sur la base d'une surveillance de réaction multiple par triple quadripôle
WO2020209469A1 (fr) Biomarqueur pour diagnostiquer chaque sous-type de neuropathie périphérique
WO2024122800A1 (fr) Nouveaux biomarqueurs pour le diagnostic et la prédiction du pronostic du cholangiocarcinome, et leur utilisation

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: 16853925

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: 16853925

Country of ref document: EP

Kind code of ref document: A1