WO2018004240A1 - Utilisation de nupr1 dans le diagnostic et le traitement de la tumeur au cerveau - Google Patents

Utilisation de nupr1 dans le diagnostic et le traitement de la tumeur au cerveau Download PDF

Info

Publication number
WO2018004240A1
WO2018004240A1 PCT/KR2017/006783 KR2017006783W WO2018004240A1 WO 2018004240 A1 WO2018004240 A1 WO 2018004240A1 KR 2017006783 W KR2017006783 W KR 2017006783W WO 2018004240 A1 WO2018004240 A1 WO 2018004240A1
Authority
WO
WIPO (PCT)
Prior art keywords
nupr1
isoform
expression
brain
brain tumor
Prior art date
Application number
PCT/KR2017/006783
Other languages
English (en)
Korean (ko)
Inventor
백선하
김요나
한범구
오미경
송민규
김현일
Original Assignee
주식회사 옵티팜
서울대학교 산학협력단
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 주식회사 옵티팜, 서울대학교 산학협력단 filed Critical 주식회사 옵티팜
Priority claimed from KR1020170081199A external-priority patent/KR101973027B1/ko
Publication of WO2018004240A1 publication Critical patent/WO2018004240A1/fr

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer

Definitions

  • the present invention relates to the use of NUPR1 in the diagnosis and treatment of brain tumors. More specifically, the present invention relates to the prevention of brain tumors comprising a biomarker composition for diagnosing or prognostic brain tumors comprising NUPR1 (NUPR1) and a NUPR1 inhibitor or A therapeutic composition.
  • NUPR1 NUPR1
  • NUPR1 inhibitor or A therapeutic composition NUPR1 inhibitor or A therapeutic composition.
  • Brain cancer is a generic term for primary brain cancers that occur in the brain tissue and the brain envelope that surrounds the brain, and secondary brain cancers that metastasize from cancers that occur in the skull or other parts of the body. Such brain cancers are often distinguished from cancers occurring in other organs.
  • the lung, stomach, breast and other cancers are limited to one or two types of organs, and their properties are the same or similar.
  • the brain develops a variety of cancers, including glioblastoma multiforme, malignant glioma, lymphadenomas, germ cell tumors, and metastatic tumors.
  • the glioma is the most frequent tumor of primary brain.
  • Meningioma is the most frequent of the brain's benign tumors with pituitary adenoma, accounting for about 15-20% of primary brain tumors. Pituitary adenoma accounts for about 10-15% of all brain tumors. Depending on the size of the tumor, it may be divided into microadenoma less than 10mm in diameter and giant adenoma greater than 10mm. Metastatic tumors account for 30 to 40% of all brain tumors, about 60% of which are observed as multiple masses, but about 40% to single masses. Therefore, it is difficult to differentiate between primary malignancies, especially glioblastoma, when they appear as a single mass. In Korea, brain cancer caused by lung cancer is the most common.
  • the 2000 HHO taxonomy includes glioblastomas, including giant cell glioblastoma and gliosarcoma.
  • the neuroeducoma (gliosarcoma) is a rare variant that contains 1.8-2.4% of glioblastoma and is clinically similar to glioblastoma. Because of the sarcomatous nature of neuroeducoma tumor cells, neuroedema is more aggressive and invasive because of its greater propensity to metastasize extracranial in neuroedema patients.
  • NUPR1 particularly NUPR1 isoform A
  • the inventors have found that NUPR1, particularly NUPR1 isoform A, is significantly increased in malignant brain tumor patients with poor prognosis among brain tumor patients, and has completed the present invention by confirming that the brain tumor can be treated by inhibiting it.
  • An object of the present invention is to provide a biomarker composition for brain tumor diagnosis or prognosis prediction comprising NUPR1 (Nuclear protein 1).
  • Another object of the present invention is a composition for diagnosing or prognosticing a brain tumor comprising a detection agent of NUPR1 (Nuclear protein 1), a kit for diagnosing or prognosticing a brain tumor comprising the composition, and for diagnosing or diagnosing a brain tumor using the composition It is to provide a method of providing information.
  • NUPR1 Nuclear protein 1
  • Still another object of the present invention is to provide a composition for preventing or treating brain tumors, including a NUPR1 (Nuclear protein 1) expression or activity inhibitor.
  • NUPR1 Nuclear protein 1
  • Still another object of the present invention is to provide a method for screening a brain tumor therapeutic agent using NUPR1 (Nuclear protein 1).
  • the present invention provides a biomarker composition for brain tumor diagnosis or prognostic prediction comprising NUPR1 (Nuclear protein 1).
  • the present invention provides a composition for diagnosing or predicting a brain tumor comprising a preparation for measuring the expression or activity level of NUPR1 (Nuclear protein 1).
  • the present invention provides a kit for diagnosing or predicting a brain tumor comprising the composition.
  • the present invention provides a method for providing information for diagnosing or predicting a brain tumor comprising a; measuring the mRNA expression of the NUPR1 gene or its protein activity level from a biological sample.
  • the present invention provides a pharmaceutical composition for preventing or treating brain tumors, including an expression or activity inhibitor of NUPR1 (Nuclear protein 1).
  • the present invention provides a food composition for preventing or improving brain tumors, including the expression or activity inhibitor of NUPR1 (Nuclear protein 1).
  • the present invention also provides a method for preventing or treating brain tumors, comprising administering an expression or activity inhibitor of NUPR1 (Nuclear protein 1) to a subject in need thereof.
  • NUPR1 Nuclear protein 1
  • the present invention comprises the steps of (a) treating a brain tumor treatment candidate material to the isolated cells; And (b) measuring the mRNA expression of the NUPR1 gene or its protein activity level in the cell.
  • NUPR1, in particular NUPR1 isoform A is significantly increased in brain tumor patients compared to healthy normal people, especially in patients with malignant brain tumors with a poor prognosis, which can be used as a biomarker for diagnosing brain tumors or predicting prognosis. have.
  • it is possible to inhibit the growth, metastasis and invasion of brain tumors through the inhibition of NUPR1, in particular NUPR1 isoform A according to the present invention it can be used as a target for the treatment of brain tumors, it can be used in the screening of future brain tumor therapeutics. .
  • 1 is a diagram confirming the protein expression of NUPR1 isoform A in normal and brain tumor-derived cells.
  • Figure 2 is a diagram showing the brain MRI pictures of patients before obtaining the GBM-28 and GBM-37 samples (from left to T1, T2, T1-enhanced MRI).
  • Figure 3 is a diagram showing the results of immunohistochemistry (H & E, GFAP, Vimentin) in brain tumor tissue obtained from GBM-28 (left) and GBM-37 (right) samples (X50).
  • FIG. 5 shows the results of performing cEA DNA array by extracting total RNA from GBM-28 and GBM-37 samples, and performing GSEA (Gene Set Enrichment Analysis) using the transcripts of each sample. It is a figure confirming whether the type is a proneural subtype or a mesenchymal subtype.
  • GSEA Gene Set Enrichment Analysis
  • Figure 6 shows the result of performing the GSEA (Gene Set Enrichment Analysis) by using the total RNA extracted from the GBM-28 and GBM-37 samples obtained cDNA microarray data, using the transcript and specific functions of each sample Figure shows the results of analyzing the correlation between the set of genes representing.
  • GSEA Gene Set Enrichment Analysis
  • FIG. 7 shows MTS proliferation assay results of primary cultured cells obtained from GBM-28 and GBM-37 samples.
  • FIG. 8 is a diagram showing the results of categorizing various disease and biological functions through IPA after extracting total RNA from GBM-28 and GBM-37 samples to obtain cDNA microarray data.
  • Figure 9 shows the extraction of total RNA from GBM-28 and GBM-37 samples, followed by RT-PCR for NUPR1 total, NUPR1 isoform A (varient 1, v.1) and NUPR1 isoform B (varient 2, v.2) mRNAs.
  • Figure 9 shows the results of analyzing the expression level.
  • FIG. 10 is a diagram showing the results of analysis of the expression of the total NUPR1 and NUPR1 isoform A (varient 1, v.1) expression by Western blot after extracting the whole protein from the GBM-28 and GBM-37 samples.
  • FIG. 11 is a diagram illustrating the expression level of NUPR1 isoform A in each group after dividing the primary glioblastoma patient group registered into TCGA (The Cancer Genome Atlas) into five subtypes.
  • FIG. 12 is a diagram showing the result of analyzing the mRNA expression of NUPR1 isoform A through RT-PCR after extracting the whole RNA from cells derived from various brain tumors.
  • FIG. 13 is a diagram showing the results of survival analysis after dividing the primary glioblastoma patient group registered in TCGA (The Cancer Genome Atlas) into high and low expression according to the degree of NUPR1 isoform A expression.
  • FIG. 14 is a first screening of MGMP methylated patients among primary glioblastoma patient groups registered in The Cancer Genome Atlas (TCGA), and then divided into high and low expression according to NUPR1 isoform A expression. This figure shows the results of the survival analysis.
  • TCGA Cancer Genome Atlas
  • FIG. 15 is a primary screening of MGMP unmethylated patients among primary glioblastoma patients registered with The Cancer Genome Atlas (TCGA), and divided into high and low expression according to the degree of NUPR1 isoform A expression. It is a figure which shows the result of performing a survival analysis.
  • TCGA Cancer Genome Atlas
  • FIG. 16 shows clinical information of 79 primary glioblastoma patients who visited Seoul National University Hospital.
  • FIG. 17 stains tissues of 79 primary glioblastoma patients who visited Seoul National University Hospital using antibodies against NUPR1 isoform A, and then divides the expression into high and low expression according to the expression level of NUPR1 isoform A. Representative tumor tissue from each group of patients is shown.
  • FIG. 18 is a diagram showing the results of a survival analysis after dividing 79 primary glioblastoma patients at Seoul National University Hospital into high and low expression according to the degree of NUPR1 isoform A expression.
  • NUPR1 isoform A isoform A
  • NUPR1 isoform B is a diagram confirming the effects on the mRNA expression of NUPR1 isoform A and NUPR1 isoform B after transducing five siRNAs (# 1 to # 5) for NUPR1 isoform A into GBM-37 cells, respectively.
  • NUPR1 isoform A protein after transfection into GBM-37 cells with five siRNAs (# 1 to # 5) for NUPR1 isoform A, respectively.
  • siRNA-NUPR1 ⁇ isoform A into GBM-37 cells.
  • siRNA-NUPR1 ⁇ isoform A into GBM-37 cells.
  • Fig. 3 shows the transcript subtype changes before and after siRNA transfection.
  • Figure 24 is transfected siRNA for NUPR1 isoform A into GBM-37 cells (GBM-37_siNUPR1a) and extracted the entire RNA to obtain cDNA microarray data, using the GSEA (Gene Set Enrichment Analysis) results
  • GSEA Gene Set Enrichment Analysis
  • FIG. 25 is transfected siRNA for NUPR1 isoform A into GBM-37 cells (GBM-37_siNUPR1a) and then extracted the entire RNA to obtain cDNA microarray data, using the GSEA (Gene Set Enrichment Analysis) results
  • Figure 3 shows the expression changes of related gene sets such as Inflammatory Response and Reactive Oxygen Species Pathway before and after siRNA transfection.
  • FIG. 26 transfected siRNA for NUPR1 isoform A into GBM-37 cells (GBM-37_siNUPR1a), extracted the entire RNA to obtain cDNA microarray data, and then used GSEA (Gene Set Enrichment Analysis) The results of the experiments were shown to confirm the expression changes of related gene sets such as Chemokine Signaling Pathway and IL6-JAK-STAT3 before and after siRNA transfection.
  • GBM-37_siNUPR1a GBM-37 cells
  • FIG. 27 is a diagram showing the results of H & E staining of brain tumor tissues generated after injection of GBM-37 cells transformed with siRNA (siRNA NUPR1v.1) for scRNA or NUPR1 isoform A into mice.
  • siRNA siRNA NUPR1v.1
  • FIG. 28 is a view showing the enlarged observation result of the brain tumor tissue of FIG. 27.
  • FIG. 29 shows the injection of GBM-37 cells transformed with siRNA (siRNA-NUPR1 ⁇ ) for scRNA (CTL) or NUPR1 isoform A into mice, followed by extraction of whole protein from the resulting brain tumor tissues, and NUPR1 via Western blot. It is a figure which shows the result of analyzing the degree of homozygous A expression.
  • the present invention provides a biomarker composition for diagnosing or predicting a brain tumor comprising NUPR1 (Nuclear protein 1).
  • NUPR1 Nuclear protein 1
  • NUPR1 Nuclear protein 1
  • the NUPR1 (Nuclear protein 1) includes NUPR1 isoform A and NUPR1 isoform B, preferably NUPR1 isoform A.
  • prognosis refers to prediction of progression and recovery of a condition and refers to prospective or preliminary evaluation.
  • the prognosis means prediction of treatment success, survival, recurrence, metastasis, etc. in the subject after treatment of brain tumor patients, and preferably means prognosis of survival.
  • a “biomarker” is a substance capable of predicting a brain tumor diagnosis or prognosis, and can be diagnosed by discriminating whether a brain tumor occurs in a biological sample, and used as a prognostic factor of a brain tumor individual having a poor prognosis. It can be used in organic biomolecules such as polypeptides, nucleic acids (e.g. mRNA, etc.), lipids, glycolipids, glycoproteins, sugars (monosaccharides, disaccharides, oligosaccharides, etc.) with significant differences between normal and brain tumor individuals. And the like.
  • the biomarker is NUPR1, which is increased in brain tumor individuals as compared to normal individuals, especially in malignant brain tumor patients with poor prognosis, and the lower the expression level, the better the prognosis factor.
  • brain tumor is a tumor occurring in the brain and the central nervous system and can be used in combination with “brain cancer", the most representative disease of the brain tumor and mixed with a high frequency "glioma” (glioma) Can be used.
  • the brain tumors collectively refer to primary brain tumors that arise from brain tissue or the brain envelope that envelops the brain, and secondary brain tumors that originate in the skull or surrounding structures, or that metastasize to the brain from other cancers of the body. Brain tumors may also be classified by cell type, morphology, cytogenetics, molecular genetics, immunologic markers, or a combination thereof, and may be classified by the World Health Organization for brain tumors. Classification classifies central nervous system tumors on a scale of malignancy based on the histological characteristics of the tumor.
  • brain tumors include glioblastomas, glioblastoma, anaplastic astrocytomas, oligodendrogliomas, ependymomas, and low-grade astrocytomas.
  • brain tumors can be divided into four subtypes in molecular aspect, and more specifically, proneural based on clinical values such as DNA methylation pattern, degree of signal transduction activation, survival and therapeutic response.
  • subtype mesenchymal subtype, classic subtype, or neural subtype.
  • the proneural subtype includes the G-CIMP subtype, and recently, there are also cases of dividing it into five subtypes.
  • the prognosis of the proneural subtype is known to be the best, and the prognosis of the mesenchymal subtype is known to be the worst, and the NUPR1, in particular, the NUPR1 isoform according to the present invention.
  • NUPR1 in particular NUPR1 isoform A
  • A is characterized by a higher expression pattern in the mesenchymal subtype than the systemic subtype. That is, NUPR1, in particular NUPR1 isoform A, according to the present invention is significantly increased in brain tumor individuals with poor prognosis among brain tumor individuals suffering from the same disease, and can not only diagnose brain tumors but also distinguish subtypes. Prognosis can be used.
  • the present invention provides a composition for diagnosing brain tumor or predicting prognosis comprising an agent for measuring the expression or activity level of Nuclear protein 1 (NUPR1).
  • NUPR1 Nuclear protein 1
  • NUPR1 particularly NUPR1 isoform A
  • Diagnosis or prognosis of brain tumors is possible by measuring mRNA expression of isoform A gene or its protein activity level.
  • "measurement of mRNA expression level of NUPR1 gene” is a process of confirming the presence and expression level of a biomarker gene in a biological sample for diagnosis or prognosis of brain tumor.
  • the amount of mRNA is measured using the agent used in the measurement method.
  • the agent for measuring the mRNA level of the NUPR1 gene in the present invention is preferably an antisense oligonucleotide, primer pair or probe, since the base sequence of the gene encoding the NUPR1 is registered in the gene bank, those skilled in the art Based on this, antisense oligonucleotides, primer pairs or probes can be designed that specifically amplify specific regions of these genes.
  • a "primer” is a nucleic acid sequence having a short free 3 'hydroxyl group, which can form complementary templates and base pairs and starts for copying the template. By a short nucleic acid sequence that functions as a point. Primers can initiate DNA synthesis in the presence of four different nucleoside triphosphates and reagents for polymerization (ie, DNA polymerase or reverse transcriptase) at appropriate buffers and temperatures.
  • the primer means all primers that can be used for amplification of the NUPR1 gene, and the sequence of the primer is not limited as long as it can bind and amplify complementarily with the gene.
  • the term "probe” refers to a nucleic acid fragment such as RNA or DNA, which corresponds to a few bases to several hundred bases, which is capable of specific binding with mRNA. You can check the presence.
  • the probe may be prepared in the form of an oligonucleotide probe, a single stranded DNA probe, a double stranded DNA probe, an RNA probe, and the like, as long as the probe may complementarily bind to the gene.
  • the sequence of is not limited.
  • Primers or probes of the invention can be synthesized chemically using phosphoramidite solid support methods, or other well known methods. Such nucleic acid sequences can also be modified using many means known in the art. Non-limiting examples of such modifications include methylation, capping, substitution with one or more homologs of natural nucleotides, and modifications between nucleotides, eg, uncharged linkages such as methyl phosphonate, phosphoester, phosphoro Amidates, carbamates, etc.) or charged linkages (eg, phosphorothioates, phosphorodithioates, etc.).
  • NUPR1 protein activity level measurement is a process of confirming the presence and expression level of the marker protein in a biological sample for the diagnosis or prognosis of brain tumor, preferably binding specifically to the NUPR1 protein Antibodies can be used to determine the amount of protein.
  • an antibody refers to a specific protein molecule directed to an antigenic site as it is known in the art.
  • an antibody refers to an antibody that specifically binds to NUPR1, which antibody is cloned into an expression vector according to a conventional method to obtain a protein encoded by the gene, and from the protein obtained It can be prepared by the phosphorus method.
  • partial peptides that may be made from such proteins, and the partial peptides of the present invention include at least seven amino acids, preferably nine amino acids, more preferably twelve or more amino acids.
  • the form of the antibody of the present invention is not particularly limited and a part thereof is included in the antibody of the present invention and all immunoglobulin antibodies are included as long as they are polyclonal antibody, monoclonal antibody or antigen-binding. Furthermore, the antibodies of the present invention also include special antibodies such as humanized antibodies.
  • Antibodies used in the present invention include functional fragments of antibody molecules as well as complete forms having two full length light chains and two full length heavy chains.
  • the functional fragment of an antibody molecule means the fragment which has at least antigen binding function, and includes Fab, F (ab '), F (ab') 2, and Fv.
  • the present invention provides a kit for diagnosing or predicting a brain tumor comprising the composition.
  • the kit for measuring the mRNA expression level of the NUPR1 gene in the present invention may be a kit containing the necessary elements necessary to perform RT-PCR.
  • the RT-PCR kits include test tubes or other suitable containers, reaction buffers, deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNase inhibitors, DEPC- It may include water (DEPC-water), sterile water and the like.
  • the kit of the present invention may be a kit including essential elements necessary for performing the microarray chip.
  • the microarray chip kit may include a substrate to which a cDNA corresponding to a gene or a fragment thereof is attached by a probe, and the substrate may include a cDNA corresponding to a quantitative control gene or a fragment thereof, using the marker of the present invention. It can be easily prepared by the manufacturing method commonly used in the art.
  • the substrate of the microarray chip is preferably coated with an active group selected from the group consisting of amino-silane, poly-L-lysine, and aldehyde, but is not limited thereto.
  • the substrate is preferably selected from the group consisting of slide glass, plastic, metal, silicon, nylon membrane and nitrocellulose membrane, but is not limited thereto.
  • the kit for measuring the NUPR1 protein activity level in the present invention may include a substrate, a suitable buffer, a secondary antibody labeled with a coloring enzyme or a fluorescent substance, and a coloring substrate for immunological detection of the antibody.
  • the substrate may be a nitrocellulose membrane, a 96 well plate synthesized with a polyvinyl resin, a 96 well plate synthesized with a polystyrene resin, a slide glass made of glass, and the like.
  • the chromophore may be a peroxidase or an alkaline force.
  • Fatase alkaline phosphatase
  • the fluorescent material may be used FITC, RITC, etc.
  • the color substrate is ABTS (2,2'-azino-bis- (3-ethylbenzothiazoline-6-sulfonic acid) ))
  • OPD o-phenylenediamine
  • TMB tetramethyl benzidine
  • the present invention provides a method for providing information for diagnosing or predicting a brain tumor, including measuring mRNA expression of a NUPR1 gene or a protein activity level thereof from a biological sample.
  • the expression of mRNA or protein activity of NUPR1, particularly NUPR1 isoform A gene is increased in tissues in brain tumor patients, especially in patients with malignant brain tumors having a short survival time due to poor prognosis. MRNA expression of the NUPR1 gene or its protein activity was found to be significantly higher. Thus, expression analysis of NUPR1 in biological samples can provide information for diagnosing brain tumors or predicting prognosis.
  • the "information providing method for diagnosis or prognosis prediction” is a preliminary step for diagnosis or prediction of prognosis and provides objective basic information necessary for the diagnosis or prognosis of brain tumor and the clinical judgment or findings of a doctor. Is excluded.
  • biological sample means a direct subject separated from an individual and measuring the expression level of a gene or protein of interest, and a sample such as tissue, cell, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid or urine. And the like.
  • a sample of the present invention may be used to predict clinical outcome after treatment, i.e., recurrence and survival prognosis, from a sample for diagnosing the onset of a brain tumor in a suspicious brain tumor or from a subject undergoing surgical or chemical treatment for a brain tumor.
  • the sample preferably brain tissue or cells isolated therefrom are not limited thereto.
  • the "analytical method for measuring mRNA expression level” includes a polymerase reaction (PCR), a reverse transcriptase polymerase reaction (RT-PCR), a competitive reverse transcriptase polymerase reaction (Competitive RT-PCR), a real time reverse transcriptase polymerase.
  • PCR polymerase reaction
  • RT-PCR reverse transcriptase polymerase reaction
  • Competitive RT-PCR competitive reverse transcriptase polymerase reaction
  • Realtime RT-PCR RNase protection assay (RPA), northern blotting, or DNA microarray analysis.
  • the "analytical method for measuring protein activity level” includes western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay, radioimmunodiffusion, and ouktero. Ouchterlony immunodiffusion, Rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, Fluorescence Activated Cell Sorter (FACS) or protein chip ( protein chip) analysis, but is not limited thereto.
  • the method of the present invention may comprise comparing the measured mRNA expression or protein activity level of the NUPR1 gene with the level measured in the control.
  • the control group includes a negative control group (normal group) or a positive control group (brain tumor patient group).
  • the positive control group may be an individual known to have a poor prognosis, for example, an individual having a history of metastasis or death after the onset of a brain tumor.
  • the positive control group may be an individual capable of confirming survival information through a known database, and preferably may be a brain tumor patient whose survival value is below average.
  • the mRNA expression of the NUPR1 gene or its protein activity level is higher than that of the negative control group (normal group)
  • the prognosis when the mRNA expression of the NUPR1 gene or its protein activity level is higher than that of the positive control group (brain tumor patient group), it may be determined that the prognosis is relatively poor and lower than that of the positive control group (brain tumor patient group). In this case, the prognosis is relatively good.
  • the present invention provides a pharmaceutical composition for preventing or treating brain tumors, including an inhibitor of NUPR1 (Nuclear protein 1) expression or activity.
  • the present invention provides a food composition for preventing or improving brain tumors, including the expression or activity inhibitor of NUPR1 (Nuclear protein 1).
  • the present invention also provides a method for preventing or treating brain tumors, comprising administering an expression or activity inhibitor of NUPR1 (Nuclear protein 1) to a subject in need thereof.
  • NUPR1 Nuclear protein 1
  • expression or activity inhibitor of NUPR1 (Nuclear protein 1) is used as a generic term for a substance that reduces the expression or activity of NUPR1, and more specifically, directly acts on NUPR1 or indirectly to its ligand. It may include any substance that reduces the expression or activity of NUPR1 by reducing the expression of NUPR1 at the transcription level or by inhibiting its activity, such as by acting.
  • the substance that inhibits the expression of NUPR1 can be used without limitation in the form of a compound, a nucleic acid, a peptide, a virus or a vector containing the nucleic acid that can target NUPR1 and inhibit the expression or activity of NUPR1.
  • Examples of substances that inhibit NUPR1 expression include siRNAs, shRNAs, miRNAs, antisense oligonucleotides, ribozymes, DNAzyme, peptide nucleic acids (PNAs) that specifically bind to NUPR1 mRNA and thereby inhibit the expression of NUPR1 mRNA.
  • examples of a substance that inhibits NUPR1 activity include an antibody or an antigen-binding fragment thereof, an aptamer, a compound thereof, or the like that specifically binds to the protein of NUPR1 to inhibit the activity of the NUPR1 protein, and preferably may be an antibody. This is not restrictive.
  • siRNA small interference RNA
  • siRNA is a nucleic acid molecule capable of mediating RNA interference or gene silencing, and refers to a small RNA fragment of 21 to 25 nucleotides in size.
  • the siRNA of the present invention may have a double-stranded structure in which the sense strand (corresponding sequence) and the antisense strand (sequence complementary to the mRNA sequence) are positioned opposite to each other and form a double-stranded structure. -complementary) can have a single chain structure with sense and antisense strands.
  • the siRNA of the present invention is not limited to the complete pairing of double-stranded RNA portions paired with RNA, but by mismatching (corresponding base is not complementary), bulge (no base corresponding to one chain), or the like. Unpaired parts may be included.
  • siRNA consisting of SEQ ID NO: 3 and 4, SEQ ID NO: 5 and 6, SEQ ID NO: 7 and 8, SEQ ID NO: 9 and 10, or SEQ ID NO: 11 and 12, the siRNA of the NUPR1 mRNA If the expression can be inhibited, the sequence is not particularly limited.
  • shRNA short hairpin RNA
  • Viruses, lentiviruses, and plasmid expression vector systems can be used to introduce and express them into cells, and these shRNAs are converted into siRNAs with the correct structure by siRNA processing enzymes (Dicer or Rnase III) present in the cells. It is well known to induce silencing of.
  • miRNA is a single-stranded RNA molecule of 21-25 nucleotides, and is a regulator that controls gene expression in eukaryotes through inhibition of the disruption or translation of target mRNAs.
  • This miRNA consists of two stages of processing.
  • the first miRNA transcript (primary miRNA) is made into a 70-90 base stem-loop structure, or pre-miRNA, by an RNaseIII type enzyme called Drosha in the nucleus, which then migrates into the cytoplasm and is called Dicer. Is cleaved into a mature miRNA of 21-25 bases.
  • the miRNA thus produced complementarily binds to the target mRNA and acts as a post-transcriptional gene suppressor, inducing translation inhibition and mRNA destabilization. miRNAs are involved in a variety of physiological phenomena and diseases.
  • antisense oligonucleotide refers to DNA or RNA or a derivative thereof containing a nucleic acid sequence complementary to the sequence of a particular mRNA, binding to the complementary sequence in the mRNA to translate the mRNA into a protein It shows an inhibitory effect.
  • aptamer means a nucleic acid molecule having binding activity to a given target molecule.
  • the aptamers may be RNA, DNA, modified nucleic acids, or mixtures thereof, and may be in linear or cyclic form, using an oligonucleotide library called systemic evolution of ligands by exponential enrichment (SELEX). It is a substance obtained by isolating oligomers which have high affinity and selectivity for binding to specific chemical or biological molecules by evolutionary methods.
  • the aptamer can specifically bind to the target and modulate the target's activity, for example, by blocking the ability of the target to function through binding.
  • prevention means any action that inhibits or delays the onset of brain tumors by administration of the composition.
  • treatment prevents the occurrence, metastasis or recurrence of brain tumors and alleviates the symptoms by administering the composition, lowers all direct or indirect pathological consequences of the disease, reduces the rate of disease progression and These include alleviating or temporarily mitigating a disease state, driving off or improving the prognosis.
  • improvement means any action that at least reduces the parameters associated with the condition being treated, for example the extent of symptoms.
  • NUPR1 inhibitor by inhibiting the expression of NUPR1, in particular NUPR1 isoform A (varient 1, v.1), not only can the growth, metastasis and invasion of brain tumors be suppressed, but also the effect of converting into a prognosis subtype is It has been confirmed that the NUPR1 inhibitor can be used as a medicine and dietary supplement for the prevention, improvement, or treatment of brain tumors.
  • composition of the present invention may further contain one or more known active ingredients having a therapeutic effect on brain tumors, together with the expression or activity inhibitor of NUPR1.
  • compositions of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.
  • Carriers, excipients and diluents which may be included in the pharmaceutical compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate and mineral oil.
  • the pharmaceutical composition of the present invention may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral dosage forms, external preparations, suppositories, and sterile injectable solutions according to conventional methods. .
  • Suitable formulations known in the art are preferably those disclosed in Remington's Pharmaceutical Science, recently, Mack Publishing Company, Easton PA.
  • when formulating the composition can be prepared using diluents or excipients, such as commonly used fillers, extenders, binders, wetting agents, disintegrants, surfactants.
  • solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may include at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin and the like are mixed and prepared.
  • excipients such as starch, calcium carbonate, Sucrose, lactose, gelatin and the like are mixed and prepared.
  • lubricants such as magnesium stearate and talc are also used.
  • Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. .
  • Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories.
  • non-aqueous solvent and suspending agent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, etc. may be used.
  • base of the suppository witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
  • administration means providing a subject with a composition of the present invention in any suitable manner.
  • Preferred dosages of the pharmaceutical compositions of the present invention depend on the condition and weight of the individual, the extent of the disease, the form of the drug, the route of administration and the duration and can be appropriately selected by those skilled in the art. The dosage does not limit the scope of the invention in any aspect.
  • the pharmaceutical composition of the present invention can be administered to a subject by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intracranial injection. However, since oral administration may denature the composition by gastric acid, oral compositions may be formulated to coat the active agent or protect it from degradation in the stomach. In addition, the composition may be administered by any device in which the active substance may migrate to the target cell.
  • the pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention or treatment of brain tumors.
  • the expression or activity inhibitor of NUPR1 of the present invention can be added to food compositions, preferably functional foods for the purpose of preventing or improving brain tumors.
  • health functional food refers to a food having a bioregulatory function, such as prevention or improvement of disease, biological defense, immunity, recovery of symptoms, inhibition of aging, and should be harmless to the human body when taken in the long term.
  • the composition When the food composition of the present invention is used as a food additive, the composition may be added as it is or used with other food or food ingredients, and may be appropriately used according to a conventional method.
  • the mixed amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment), and there is no particular limitation on the type of food.
  • the food composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonic acid. Carbonating agents used in beverages and the like.
  • the present invention comprises the steps of (a) treating a brain tumor treatment candidate to isolated brain tumor cells; And (b) measuring the mRNA expression of the NUPR1 gene or its protein activity level in the brain tumor cells.
  • the candidate of step (a) is a substance that is expected to be able to treat brain tumors or an unknown substance that is expected to improve the prognosis, and includes a compound, protein or natural product extract, but is not limited thereto. It is not.
  • the method of the present invention (c) when the mRNA expression or protein activity level of the NUPR1 gene measured in step (b) shows a lower level than the brain tumor cells not treated with the candidate material, the candidate material as a brain tumor treatment agent Determining that it can be used.
  • the method of measuring the mRNA or protein level of the gene is the same as described above.
  • NUPR1 isoform A (v.1) protein.
  • SVG p12 used as a control means normal cells as human derived astroglia.
  • a primary antibody specific for NUPR1 isoform A a polyclonal antibody prepared in Example 2-4 described below was used.
  • ⁇ -actin was used as a housekeeping protein for quantification, and brain tumor patient information is shown in Table 1 below. The results are shown in FIG.
  • GBM-12 Male 46 Glioblastoma (grade IV) GBM-14 Male 52 Anaplastic Astrocytoma (grade III) GBM-15 Female 37 Glioblastoma (grade IV) GBM-30 Female 46 Glioblastoma (grade IV)
  • NUPR1 isoform A protein is hardly expressed in the SVG p12 sample which is normal cells, whereas NUPR1 isoform A protein is expressed in the GBM-12, GBM-14, GBM-15 and GBM-30 samples of brain tumor patients. It was confirmed that this is markedly high.
  • NUPR1 isoform A (varient 1, v.1) is expressed higher in the brain tumor patient group than healthy normal group, it was confirmed that it can be used as a diagnostic marker for brain tumors.
  • Brain tumor tissue samples were obtained from primary glioblastoma patients (Glioblastoma, GBM-28) and secondary neuroeducoma patients (Gliosarcoma, GBM-37), followed by primary culture.
  • GBM-28 and GBM-37 are samples derived from the same patient, and GBM-37 is a sample at the time of recurrence after 3 months after tumor removal surgery and concurrent chemoradiotherapy after obtaining the GBM-28 sample. .
  • Brain MRI photographs of patients before sample acquisition are shown in FIG. 2 (T1, T2, T1-enhanced MRI from left).
  • FBS fetal bovine serum, Gibco Corp., 16000, Grand Island, NY, USA
  • DMEM medium Dulbecco's modified Eagle medium, WelGENE, LM001-05, Korea), including Island, NY, USA
  • H & E GFAP, Vimentin, X50
  • morphology observations were performed on cells obtained from patients with primary glioblastoma (GBM-28) and patients with secondary neuroedema (GBM-37). 3 and 4 are shown.
  • GBM-28 cells isolated from primary glioblastoma patients show typical characteristics of glioblastoma
  • GBM-37 cells isolated from secondary neuroedema patients The right side of each figure was confirmed that the GFAP staining and distinguished from the normal glioblastoma, such as showing a specific form (fascicular arranged spindle cells).
  • cDNA microarray was performed. Transcript analysis was performed by oligonucleotide microarray analysis using MA-human Agilent 44k (Agilent Technologies), and one-channel microarray data was analyzed by global median normalization method using GeneSpring GX 7.3 (EBIOGEN, Seoul, Korea). Using the analyzed cDNA microarray data, Gene Set Enrichment Analysis, Broad Institute, USA (GSEA) was performed to select a gene set specifically correlated in the transcripts of cells derived from each brain tumor patient.
  • GSEA Gene Set Enrichment Analysis, Broad Institute, USA
  • GSEA was run through the javaGSEA desktop application (GSEA v2.1.) And the gene set was downloaded from a database called MSigDb (http://www.broadinstitute.org/gsea/msigdb/). GSEA produces a enrichment score, a normalized enrichment score, a nominal p-value, and a false discovery rate (q-value). Only gene sets with p-values less than 0.05 were selected to compare GBM-28 transcripts and GBM-37 transcripts. The results are shown in FIGS. 5 and 6.
  • GBM-28 shows a proneural subtype with a prognosis.
  • GBM-37 confirms that the prognosis represents a very poor Mesenchymal subtype.
  • MTS cell proliferation assays were performed on cells isolated from primary glioblastoma patients (GBM-28) and secondary neuroeducoma patients (GBM-37). . First, they seeded the cells in each 96-well culture plate with 1.0 ⁇ 10 4 cells / well ratios. After cell culture for a period of time, CellTiter 96® AQueous One Solution cell proliferation assay (Promega, 3580) solution was added to each well. The absorbance at 490 nm was then measured using Infinite M200 Pro (Tecan). The results are shown in FIG.
  • GBM-37 cells isolated from secondary neuroeducoma patients were found to have higher cell proliferation than GBM-28 cells isolated from primary glioblastoma patients.
  • Example 2-1 a bioinformatics program called IPA was used.
  • the cDNA microarray data obtained in Example 2-1 was analyzed using a tool called Core analysis of IPA, and transcriptome analysis was performed using a detailed program technique called “Disease and bio functions” provided by Core analysis. .
  • the results are shown in FIG.
  • GBM-37 As shown in FIG. 8, as a result of classifying the transcripts isolated from primary glioblastoma patients (GBM-28) and secondary neuroeducoma patients (GBM-37), GBM-37 was more malignant than GBM-28. It was again confirmed that the island had high properties. Through the above experimental results, it can be seen that the subjects corresponding to GBM-37 recurred as a more malignant brain tumor, that is, a secondary neuroedema with a poor prognosis after chemotherapy.
  • NUPR1 isoform A (varient 1, v.1) was found to be expressed at a significantly high level in GBM-37.
  • NUPR1 isoform A (varient 1, v.1) is found to be higher in malignant tumor patients with a poorer prognosis than among brain tumors.
  • total protein was obtained using RIPA buffer according to a known method in brain tumor patient derived cells obtained from primary glioblastoma patient (GBM-28) and secondary neuroeducoma patient (GBM-37).
  • the protein samples were subjected to Western blot using antibodies against NUPR1 as a whole and NUPR1 isoform A (varient 1, v.1).
  • ⁇ -actin was used as a housekeeping protein for quantification.
  • the antibody against NUPR1 isoform A (varient 1, v.1) was prepared by the following procedure. First, cysteine was added to the N-terminus of the NUPR1a specific peptide (18 amino acids 38 to 55 positions from the N-terminus of NUPR1a (NP_001035948.1)), and then conjugated to KLH. The fusion peptide was immunized subcutaneously with 0.5 mg each of two rabbits using PBS and Freund's adjuvant. The same amount of fusion peptide was immunized subcutaneously three times at two week intervals after primary immunization. One week after the final immunization, blood was collected and polyclonal antibodies were obtained using Protein A resin. The polyclonal antibody against NUPR1 isoform A (variant 1, v.1) obtained through the above procedure was used for experiments after confirming specificity through assays with antibodies produced by expressing recombinant proteins of NUPR1a and NUPR1b.
  • the total NUPR1 protein expression in the GBM-28 and GBM-37 samples did not show a specific difference, but the NUPR1 isoform A (varient 1, v.1) protein was compared with the GBM-28 sample. It was found to be significantly higher in 37 samples. In other words, NUPR1 isoform A (v.1) protein was expressed higher in malignant tumor patients with poorer prognosis than among brain tumors as in mRNA expression analysis.
  • the primary glioblastoma patient group enrolled in the Cancer Genome Atlas was divided into five subtypes (Proneural, Classical, Neural, Mesenchymal, G-CIMP), followed by NUPR1 isoform A (ID: ENST00000395641).
  • the expression level was analyzed.
  • the G-CIMP subtype belongs to the Proneural subtype and is a subtype known to have a good prognosis. This subtype is known to be closely associated with the prognostic factor of a patient called the IDH1 / 2 mutation. The results are shown in FIG.
  • NUPR1 isoform A (varient 1, v.1) is expressed at a higher level in malignant brain tumors classified as mesenchymal subtypes with the poorest prognosis among brain tumors. It was confirmed that it can be used not only for the diagnosis of brain tumors but also for the diagnosis of subtypes of brain tumors.
  • RNA was isolated from brain tumor patient derived cells obtained from brain tumor patients with different diagnosis names and survival rates. RT-PCR was performed in the same manner as in 2-3. Specific patient information is shown in Table 2, and the experimental results are shown in FIG. U87 is a human primary glioblastoma cell line sold by the American Type Culture Collection (ATCC).
  • ATCC American Type Culture Collection
  • the mRNA expression level of NUPR1 isoform A (varient 1, v.1) was significantly higher in the GBM-37 sample derived from the patient with extremely poor prognosis due to only 7 months of survival compared to the sample derived from other brain tumor patients. It was confirmed.
  • the primary glioblastoma patient group registered in The Cancer Genome Atlas (TCGA) was divided into high and low expression according to the degree of NUPR1 isoform A expression, and then survival analysis was performed. More specifically, an RNA-sequencing dataset was used, and the ID of NUPR1 isoform A was identified as ENST00000395641 (uc002dqd.2). Using the ID, the expression value of NUPR1 isoform A was confirmed for each patient, and then classified into high and low expression according to the expression level of NUPR1 isoform A and survival analysis was performed.
  • MGMT methylation which is used as a prognostic factor for glioblastoma patients, was classified primarily in the same patient group, and then expression level and survival analysis of NUPR1 isoform A were performed. The results are shown in FIGS. 13 to 15.
  • NUPR1 isoform A (varient 1, v.1) shows a significant correlation with the survival rate, which can be used as a marker for predicting the prognosis of brain tumor patients. Confirmed.
  • siRNAs capable of specifically inhibiting the NUPR1 isoform A were prepared (Genolution). After culturing GBM-37 cells for 24 hours, siRNA was transfected into cells using LipofectamineTM RNAiMAX (Invitrogen). 24 hours after transfection, total RNA and protein were isolated from each cell and RT-PCR and Western blots were performed according to known methods. The siRNA sequences are shown in Table 3, with RT-PCR results shown in FIG. 19 and Western blot results shown in FIG. 20.
  • NUPR1 ⁇ As shown in FIG. 19, it was confirmed that all five siRNAs for NUPR1 isoform A did not affect NUPR1 isoform B (NUPR1 ⁇ ), but specifically downregulated mRNA expression of NUPR1 isoform A (NUPR1 ⁇ ).
  • NUPR1 ⁇ protein expression of NUPR1 isoform A (NUPR1 ⁇ ) is also down-regulated by the transfection of siRNA against NUPR1 isoform A.
  • GBM-37 cells isolated from malignant brain tumor patients were transfected with siRNA (NUPR1 V1 # 2) prepared in Example 4-1, followed by MTS cell proliferation assay.
  • siRNA NUPR1 V1 # 2
  • MTS cell proliferation assay MTS cell proliferation assay.
  • scRNA scRNA
  • each cell was seeded in a 96-well culture plate at a rate of 1.0 ⁇ 10 4 cells / well. After cell culture for a period of time, CellTiter 96® AQueous One Solution cell proliferation assay (Promega, 3580) solution was added to each well. The absorbance at 490 nm was then measured using Infinite M200 Pro (Tecan). The results are shown in FIG.
  • GBM-37 cells isolated from malignant brain tumor patients were transfected with siRNA (NUPR1 V1 # 2) prepared in Example 4-1, followed by a transwell migration assay.
  • siRNA NUPR1 V1 # 2
  • scRNA transfected with scRNA were used.
  • each cell was released in serum-free medium, and the cells were incubated for 24 hours in transwell inserts of 24-well plates (BD Biosciences, # 3422). Then medium with 10% FBS (fetal bovine serum) was added to the lower chamber of the plate as a chemoattractant. After incubating the cells for 24 hours, non-migrated cells were removed using a cotton swab. The remaining metastatic cells were stained using crystal violet (0.5% in 20% methanol) and then harvested.
  • Invasion assays were performed under the same conditions as the transwell transfer assay, and growth factor-reduced matrigel-coated insert wells (BD Biosciences, # 356234) were used. Cell metastasis and invasion values were calculated by dividing the microscopic field into quarters and calculating the average number of cells. The results are shown in FIG. 22.
  • siRNA against NUPR1 isoform A significantly inhibited metastasis and invasion of GBM-37 cells.
  • GBM-37 before transfection showed a Mesenchymal subtype with poor prognosis (see FIG. 5), but it was possible to inhibit the expression of NUPR1 isoform A using siRNA against NUPR1 isoform A.
  • the prognosis showed good proneural subtype. This is a result showing that the properties of cancer cells have been changed to a proneural subtype with a better prognosis by siRNA for NUPR1 isoform A.
  • FIGS. 24 to 26 The results are shown in FIGS. 24 to 26.
  • the left side is the GBM-37 sample and the right side is the GBM-37 sample after transformation with siRNA for NUPR1 isoform A.
  • Metastasis related gene set was down-regulated in the GBM-37 sample (GBM-37_siNUPR1a) after transformation with siRNA for NUPR1 isoform A compared to wild type.
  • NUPR1 V1 # 2 siRNA (NUPR1 V1 # 2) or scRNA-transformed GBM-37 cells (2 ⁇ 10 6 cells) for NUPR1 isoform A were each examined for 5 weeks of athymic mice (BALB / c nu / nu) (Central Laboratory Animal). , Inc, Korea). Intracellular injection was performed in the left forehead and the coordinates were set 2 mm laterally and 0.5 mm forward from bregma. Tumors formed 7 days after cell injection were extracted and immediately stored in Bouin's fixative for 2-3 hours. Deparaffinized tumor tissue was H & E stained. Images of the stained tissue were obtained through Olympus whole-slide scanner with OlyVIA software (Olympus Life Science, Center Valley, PA, USA). The results are shown in FIGS. 27 and 28.
  • tumor size was decreased in the group to which GBM-37 cells transfected with siRNA for NUPR1 isoform A (siRNA NUPRv.1).
  • NUPR1, in particular NUPR1 isoform A is significantly increased in brain tumor patients compared to healthy normal persons, especially in patients with malignant brain tumors with a poor prognosis, which is a biomarker for diagnosing brain tumors or predicting prognosis. Can be utilized as In addition, it is possible to inhibit the growth, metastasis and invasion of brain tumors through the inhibition of NUPR1, in particular NUPR1 isoform A according to the present invention, it can be used as a target for the treatment of brain tumors, it can be used in the screening of future brain tumor therapeutics. .

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Hematology (AREA)
  • Mycology (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Hospice & Palliative Care (AREA)
  • Cell Biology (AREA)
  • Oncology (AREA)
  • Food Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne l'utilisation d'une protéine nucléaire (1) (NUPR1) dans le diagnostic et le traitement des tumeurs au cerveau et, plus particulièrement, une composition de biomarqueur comprenant NUPR1 pour le diagnostic des tumeurs au cerveau ou pour prédire le prognostic et une composition comprenant un inhibiteur NUPR1 pour prévenir ou traiter les tumeurs au cerveau. Le niveau de NUPR1, particulièrement l'isoforme (A) de NUPR1, selon la présente invention, est significativement accru chez les patients présentant des tumeurs au cerveau et remarquablement en outre chez les patients présentant des tumeurs malignes au cerveau d'un prognostic médiocre, comparé aux personnes normales saines, de telle sorte que NUPR1 peut être utilisé comme biomarqueur pour le diagnostic des tumeurs au cerveau ou pour la prédiction d'un prognostic. De plus, NUPR1, particulièrement l'isoforme (A) de NUPR1, selon la présente invention, peut être utilisé comme cible de traitement de la tumeur au cerveau et également, dans le futur, dans le criblage d'un agent thérapeutique pour les tumeurs au cerveau dans la mesure où son inhibition peut supprimer la croissance, la métastase et l'invasion de tumeurs au cerveau.
PCT/KR2017/006783 2016-06-27 2017-06-27 Utilisation de nupr1 dans le diagnostic et le traitement de la tumeur au cerveau WO2018004240A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR20160080044 2016-06-27
KR10-2016-0080044 2016-06-27
KR1020170081199A KR101973027B1 (ko) 2016-06-27 2017-06-27 뇌종양의 진단 및 치료에서의 nupr1의 용도
KR10-2017-0081199 2017-06-27

Publications (1)

Publication Number Publication Date
WO2018004240A1 true WO2018004240A1 (fr) 2018-01-04

Family

ID=60787344

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2017/006783 WO2018004240A1 (fr) 2016-06-27 2017-06-27 Utilisation de nupr1 dans le diagnostic et le traitement de la tumeur au cerveau

Country Status (1)

Country Link
WO (1) WO2018004240A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112662775A (zh) * 2021-01-18 2021-04-16 中国医科大学附属第一医院 胶质瘤标志物spc25及其应用
CN110205320B (zh) * 2019-06-18 2023-06-23 中国医科大学附属盛京医院 一种lncRNA分子linc00998及其在胶质瘤治疗/预后评估中的应用
EP4260861A1 (fr) * 2022-04-13 2023-10-18 Seoul National University R & DB Foundation Composition pharmaceutique anticancéreuse comprenant un inhibiteur de l'isoforme a de nupr1

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010064702A1 (fr) * 2008-12-05 2010-06-10 国立大学法人 東京大学 Biomarqueur pour prédire un pronostic de cancer
WO2012055565A1 (fr) * 2010-10-29 2012-05-03 Institut National De La Sante Et De La Recherche Medicale (Inserm) Nouveau marqueur de tumeurs du sein du sous-type luminal b

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010064702A1 (fr) * 2008-12-05 2010-06-10 国立大学法人 東京大学 Biomarqueur pour prédire un pronostic de cancer
WO2012055565A1 (fr) * 2010-10-29 2012-05-03 Institut National De La Sante Et De La Recherche Medicale (Inserm) Nouveau marqueur de tumeurs du sein du sous-type luminal b

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHOWDHURY, U. R.: "Emerging role of nuclear protein 1 (NUPR1) in cancer biology", CANCER AND METASTASIS REVIEWS, vol. 28, no. 1-2, 20 January 2009 (2009-01-20), pages 225 - 232, XP019671835 *
GUO, X. ET AL.: "Lentivirus-mediated RNAi knockdown of NUPR1 inhibits human nonsmall cell lung cancer growth in vitro and in vivo", THE ANATOMICAL RECORD, vol. 295, no. 12, 7 September 2012 (2012-09-07), pages 2114 - 2121, XP055600863, DOI: 10.1002/ar.22571 *
LEE, Y. K. ET AL.: "Identification of a mitochondrial defect gene signature reveals NUPR1 as a key regulator of liver cancer progress ion", HEPATOLOGY, vol. 62, no. 4, 7 August 2015 (2015-08-07), pages 1174 - 1189, XP055600869, DOI: 10.1002/hep.27976 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110205320B (zh) * 2019-06-18 2023-06-23 中国医科大学附属盛京医院 一种lncRNA分子linc00998及其在胶质瘤治疗/预后评估中的应用
CN112662775A (zh) * 2021-01-18 2021-04-16 中国医科大学附属第一医院 胶质瘤标志物spc25及其应用
EP4260861A1 (fr) * 2022-04-13 2023-10-18 Seoul National University R & DB Foundation Composition pharmaceutique anticancéreuse comprenant un inhibiteur de l'isoforme a de nupr1

Similar Documents

Publication Publication Date Title
US20140235486A1 (en) Methods and Compositions for the Treatment and Diagnosis of Breast Cancer
WO2018004240A1 (fr) Utilisation de nupr1 dans le diagnostic et le traitement de la tumeur au cerveau
JP2010178650A (ja) 固形癌の再発予測のための試験方法および再発予防剤
WO2016152352A1 (fr) Biomarqueur spécifique du mélanome et son utilisation
EP2118318B1 (fr) Mutations gnaq dans un melanome
CN106636443B (zh) Dnah14基因在制备肿瘤诊治产品中的应用
KR102216590B1 (ko) 췌장 신경내분비종양의 진단을 위한 조성물
WO2020080861A1 (fr) Composition de traitement du cancer gastrique comprenant un inhibiteur de syt11 en tant qu'ingrédient actif
WO2022019604A1 (fr) Biomarqueur spécifique du cancer du foie et utilisation associée
KR102018899B1 (ko) p53-비돌연변이 암에 대한 암 마커 유전자 및 치료제 스크리닝 방법
WO2017164568A1 (fr) Procédé de prédiction du taux de survie et du pronostic de patient présentant un cancer de l'œsophage en mesurant le niveau d'expression de protéine de la sous-unité v-atpase v1e1
WO2014014157A1 (fr) Utilisation de l'adcy3 pour diagnostiquer et traiter le cancer de l'estomac
WO2022203314A2 (fr) Composition pour le diagnostic différentiel d'une tumeur maligne des gaines nerveuses périphériques
Zhong et al. Aberrant DNA methylation of synaptophysin is involved in adrenal cortisol-producing adenoma
KR101973027B1 (ko) 뇌종양의 진단 및 치료에서의 nupr1의 용도
KR102435516B1 (ko) 줄기세포의 노화 진단용 조성물
WO2018174506A1 (fr) Procédé de prédiction de la susceptibilité au traitement par sorafénib à l'aide d'un gène sulf2, et composition pour le traitement du cancer comprenant un inhibiteur de sulf2
KR101916898B1 (ko) 비만 치료 또는 예방용 약학 조성물
CN109371136B (zh) 一种与肺腺癌相关的lncRNA及其应用
US20180057887A1 (en) Her2-regulated rna as a diagnostic and therapeutic targets in her2+ breast cancer
KR20220062164A (ko) Glis1 발현 또는 활성 억제제를 포함하는 난소암 전이 억제용 약학적 조성물
WO2020117002A1 (fr) Méthode pour établir le pronostic d'un carcinome épidermoïde oesophagien
WO2011122859A2 (fr) Composition permettant de prédire la récidive d'une tumeur cérébrale et les chances de survie et kit la contenant
WO2023244016A1 (fr) Composition pour la prévention ou le traitement de l'arthrose et son utilisation
CN116463424B (zh) 生物标志物在黑色素瘤治疗、预防或预后预测中的应用

Legal Events

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

Ref document number: 17820516

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: 17820516

Country of ref document: EP

Kind code of ref document: A1