Composition for the differential diagnosis of malignant peripheral nerve schwannoma

Abstract

The present invention relates to a composition for differential diagnosis of malignant peripheral nerve sheath tumor (MPNST), a differential diagnosis kit containing the same, a method for providing information necessary for differential diagnosis of malignant peripheral nerve sheath tumor using the same, a screening method for substances for preventing, improving and treating malignant peripheral nerve sheath tumor, etc. By using a composition capable of detecting the protein marker of the present invention, it is possible to predict and diagnose the occurrence of malignant tumors in patients with neurofibromatosis type 1.

Description

Technical Field

[0001] The present invention relates to a composition for differential diagnosis of malignant peripheral nerve sheath tumor (MPNST), a differential diagnosis kit containing the same, a method for providing information necessary for differential diagnosis of malignant peripheral nerve sheath tumor using the same, a method for screening substances for prevention, improvement and treatment of malignant peripheral nerve sheath tumor, and the like.

[0002] The present invention claims priority based on Korean Patent Application No. 10-2021-0037912 filed on March 24, 2021 and Korean Patent Application No. 10-2022-0034295 filed on March 18, 2022, and all the contents disclosed in the specifications and drawings of the above applications are incorporated herein by reference.

Background Art

[0003] Neurofibromatosis type I (hereinafter referred to as "NF-1") was first described by Fredrich von Recklinghausen in 1882 and is one of the most common genetic diseases. NF-1 is inherited through autosomal dominant inheritance and affects 1 in 3,000 people worldwide. NF-1 is characterized by multiple café-au-lait spots on the skin and cutaneous neurofibromas. In addition, various levels of nervous system, skeletal system expression and neoplastic expression may affect patients. NF-1 patients are at risk of developing central and peripheral nervous system tumors including plexiform neurofibromas (PN; 27%), optic nerve gliomas (Optic nerve glioma; 15-20%), pheochromocytomas (1%), and malignant peripheral nerve sheath tumors (MPNST; 8-13%). This is about 1,000 times higher than that of the general population.

[0004] Malignant peripheral nerve sheath tumors (MPNSTs) are one of the complications that have the greatest impact on the survival rate of NF-1 patients. They are known to result from malignant deformation of plexiform neurofibromas, which are benign nerve sheath tumors, and should be suspected when accompanied by severe pain, rapid increase in fibroma size, and neurological symptoms.

[0005] To date, 18F-FDG-PET (18F-fluorodeoxyglucose positron emission computerized tomography) has been used to differentiate between benign tumors and MPNSTs, and if malignancy is suspected, MPNSTs are diagnosed based on histological findings.

[0006] Most MPNSTs histologically present as bundles of spindle cells, with cells showing advanced nuclear differentiation and exhibiting infiltration into surrounding tissues. S100 protein staining is used for the diagnosis of MPNST, but staining is benign in 50-90% of MPNST cases. In addition, Leu-7 protein and myelin basic protein staining are benign in approximately 50% of MPNST cases. Vimentin is also a pathological marker used for the diagnosis of MPNST, but it is specific to spindle cells and is therefore limited in its specific diagnosis of MPNST. Consequently, there is a need for pathological indicators that can clearly determine the progression of MPNST in NF-1.

[0007] MPNST is a disease that requires surgical resection after diagnosis. Approximately 50% of patients are at risk of recurrence after surgery, and if complete resection is not achieved during surgery, chemotherapy and radiation therapy may be necessary. Despite these challenges, the five-year survival rate for MPNST patients remains at only 20-50%.

[0008] To date, there are no therapies that target MPNST. Therefore, finding biomarkers that can diagnose MPNST early from blood or urine is thought to contribute to improving patient survival rates. [Overview of the project] [Problems that the invention aims to solve]

[0009] Therefore, the inventors conducted extensive quantitative and qualitative proteomic analyses using malignant peripheral nerve sheath tumor (MPNST) and benign neurofibroma tissues, and as a result identified proteins specifically expressed in MPNST tissue, thus completing the present invention.

[0010] Therefore, the object of the present invention is to provide a composition for the differential diagnosis of malignant peripheral nerve schwannoma (MPNST).

[0011] Another object of the present invention is to provide a kit for the differential diagnosis of malignant peripheral nerve schwannomas comprising the composition according to the present invention.

[0012] Another object of the present invention is to provide a method for providing information necessary for the differential diagnosis of malignant peripheral nerve schwannomas.

[0013] A further object of the present invention is to provide compositions for the prevention, improvement, or treatment of malignant peripheral nerve schwannomas.

[0014] A further object of the present invention is to provide a method for screening substances for the prevention, improvement, or treatment of malignant peripheral nerve schwannomas.

[0015] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the following description. [Means for solving the problem]

[0016] To achieve the objectives of the present invention, the present invention provides a composition for the differential diagnosis of malignant peripheral nerve sheath tumor (MPNST), comprising as an active ingredient a formulation for measuring the mRNA expression level of at least one protein selected from the group consisting of endosialin, SPARC-like protein 1, and neutrophil collagenase.

[0017] In one embodiment of the present invention, the composition may be used to distinguish and diagnose malignant peripheral nerve schwannoma from neurofibromatosis type I.

[0018] In other embodiments of the present invention, the formulation for measuring the expression level of the protein may be, but is not limited to, an antibody or aptamer specific to the protein.

[0019] In yet another embodiment of the present invention, the formulation for measuring the mRNA expression level may be, but is not limited to, a primer set or probe that specifically binds to the mRNA.

[0020] Furthermore, the present invention provides a kit for the differential diagnosis of malignant peripheral nerve schwannoma, comprising the composition according to the present invention.

[0021] In one embodiment of the present invention, the kit may include instructions indicating that if the expression level of at least one protein or its mRNA selected from the group consisting of endosialin, SPARC-like protein 1, and neutrophil collagenase is detected, or if their expression levels are higher than those of a benign neurofibroma sample, it is determined to be a malignant peripheral schwannoma.

[0022] Furthermore, the present invention provides a method for providing information necessary for the differential diagnosis of malignant peripheral nerve schwannoma, comprising the step of measuring the mRNA expression level of at least one protein selected from the group consisting of endosialin, SPARC-like protein 1, and neutrophil collagenase in a biological sample isolated from a subject.

[0023] Furthermore, the present invention provides a method for the differential diagnosis of malignant peripheral nerve sheath tumors, comprising the step of measuring the mRNA expression level of at least one protein selected from the group consisting of endosialin, SPARC-like protein 1, and neutrophil collagenase in a biological sample isolated from a subject.

[0024] In one embodiment of the present invention, the method may further include the step of diagnosing a malignant peripheral schwannoma if the expression of at least one protein or its mRNA selected from the group consisting of endosialin, SPARC-like protein 1, and neutrophil collagenase is detected, or if the expression levels thereof are higher than the expression levels in biological samples isolated from patients with neurofibromatosis type 1.

[0025] In another embodiment of the present invention, the subject may be a patient suspected of having malignant peripheral nerve schwannoma or a patient diagnosed with neurofibromatosis type 1.

[0026] In still other embodiments of the present invention, the biological sample may be at least one selected from the group consisting of blood, whole blood, plasma, urine, tissue, cells, organs, bone marrow, fine needle aspiration specimens, fine needle washings, core needle biopsy specimens, and saliva, but is not limited thereto.

[0027] In still other embodiments of the present invention, the protein expression level may be measured by at least one method selected from the group consisting of Western blot, ELISA, radioimmunoassay, radioimmunodiffusion, Ouchterlony immunodiffusion, Rocket immunoelectrophoresis, immunostaining, immunoprecipitation analysis, complement fixation analysis, mass spectrometry, FACS, and protein chips, but is not limited thereto.

[0028] In still other embodiments of the present invention, the mRNA expression level may be measured by at least one method selected from the group consisting of PCR, RNase protection assay, northern blotting, southern blotting, in situ hybridization, and DNA chips, but is not limited thereto.

[0029] The present invention also provides a pharmaceutical composition for preventing or treating malignant peripheral nerve sheath tumors, comprising, as an active ingredient, an inhibitor of the activity of at least one signal transduction system protein selected from the group consisting of TGF-β signal transduction system, ILK signal transduction system, actin-cytoskeleton signal transduction system, and RhoGDI signal transduction system, or an inhibitor of the expression of the gene encoding the protein.

[0030] Furthermore, the present invention provides a method for preventing or treating malignant peripheral nerve schwannoma, comprising the step of administering an inhibitor of the activity of at least one signaling pathway protein selected from the group consisting of the TGF-β signaling pathway, the ILK signaling pathway, the actin-cytoskeleton signaling pathway, and the RhoGDI signaling pathway, or a gene expression inhibitor encoding the said protein, to an individual in need.

[0031] Furthermore, the present invention provides an inhibitor of the activity of at least one signaling pathway protein selected from the group consisting of the TGF-β signaling pathway, the ILK signaling pathway, the actin-cytoskeleton signaling pathway, and the RhoGDI signaling pathway, or an inhibitor of the expression of the gene encoding said protein, for use in the prevention or treatment of malignant peripheral nerve schwannomas.

[0032] Furthermore, the present invention provides a health functional food composition for the prevention or improvement of malignant peripheral nerve schwannoma, comprising, as an active ingredient, an inhibitor of the activity of at least one signaling pathway protein selected from the group consisting of the TGF-β signaling pathway, the ILK signaling pathway, the actin-cytoskeleton signaling pathway, and the RhoGDI signaling pathway, or an inhibitor of the expression of the gene encoding said protein.

[0033] In one embodiment of the present invention, the signal transduction pathway protein may be a kinase, but is not limited thereto.

[0034] In other embodiments of the present invention, the kinase may be, but is not limited to, a kinase encoded by at least one gene selected from the group consisting of CDK4, STK10, CDK5, MAP2K1, ALPK3, PNKP, PDK3, PHKG1, CFL1, and PACSIN3.

[0035] In yet another embodiment of the present invention, the protein activity inhibitor may be, but is not limited to, at least one selected from the group consisting of peptides, antibodies, aptamers, and compounds that specifically bind to the protein.

[0036] In yet another embodiment of the present invention, the inhibitor of the activity of the signaling pathway protein is selumetinib, alvocidib, purvalanol, palbociclib, ribociclib, abemaciclib, fostamatinib, binimetinib, radicicol, AZD-8330, dihydrolipoic acid At least one selected from the group consisting of acid, alsterpaullone, hymenialdisine, indirubin-3'-monoxime, olomoucine, SU9516, and 6-phenyl[5H]pyrrolo[2,3-B]pyrazine may be present, but is not limited to.

[0037] In yet another embodiment of the present invention, the gene expression inhibitor may be, but is not limited to, at least one selected from the group consisting of miRNA, siRNA, shRNA, ribozyme, DNAzyme, PNA (peptide nucleic acid), and antisense oligonucleotide that bind complementarily to the mRNA of the gene.

[0038] The present invention also provides a method for screening for substances for the prevention, improvement, or treatment of malignant peripheral nerve schwannomas, comprising the steps of: (a) contacting isolated cells or tissues with a candidate substance; (b) measuring the activity of at least one signaling pathway protein selected from the group consisting of the TGF-β signaling pathway, the ILK signaling pathway, the actin-cytoskeleton signaling pathway, and the RhoGDI signaling pathway, or the expression level of a gene encoding the protein, in the cells or tissues; and (c) determining that the candidate substance is a substance for the prevention, improvement, or treatment of malignant peripheral nerve schwannomas if the activity of the protein or the expression level of the gene encoding the protein decreases in the cells or tissues that have been in contact with the candidate substance compared to the level before contact with the candidate substance.

[0039] In one embodiment of the present invention, the signal transduction pathway protein may be a kinase, but is not limited thereto.

[0040] In other embodiments of the present invention, the kinase may be, but is not limited to, a kinase encoded by at least one gene selected from the group consisting of CDK4, STK10, CDK5, MAP2K1, ALPK3, PNKP, PDK3, PHKG1, CFL1, and PACSIN3.

[0041] In yet another embodiment of the present invention, the isolated cells or tissue may be, but are not limited to, nerve cells or nerve tissue. [Effects of the Invention]

[0042] The present invention relates to a composition for the differential diagnosis of malignant peripheral nerve sheath tumor (MPNST), and the composition, which can detect the protein marker of the present invention, can be used to predict and diagnose the presence or absence of malignant tumors in patients with neurofibromatosis type 1, and to utilize them as therapeutic targets. [Brief explanation of the drawing]

[0043] [Figure 1a-1b] Figures 1a and 1b are time-series flowcharts of quantitative proteomic analysis using benign neurofibroma (NF) and malignant peripheral nerve schwannoma (MPNST) tissues. [Figure 2] Figure 2 is a diagram illustrating the proteome data analysis method. [Figure 3] Figure 3 is a protein abundance graph showing the quantitative results of proteins in benign neurofibroma (NF) and malignant peripheral nerve schwannoma (MPNST) tissues. [Figure 4] Figure 4 shows the results of qualitative proteomic analysis for two groups, neurofibroma (NF) and malignant peripheral nerve schwannoma (MPNST), using Ven diagram analysis. [Figures 5a-5b] Figures 5a and 5b show the results of a protein analysis that showed a more than 10-fold increase in expression in benign neurofibroma (NF) and malignant peripheral nerve schwannoma (MPNST) tissues. Figure 5a shows the results of selecting proteins expressed in extracellular exosomes, while Figure 5b shows the results of selecting proteins that degrade the extracellular matrix. [Figures 6a-6j]Figures 6a to 6j are graphs showing the results of investigating the expression patterns of S-100 isomorphic proteins and vimentin in benign neurofibroma (NF) and malignant peripheral nerve schwannoma (MPNST) tissues. Figure 6a shows the results for S100-A1 protein, Figure 6b shows the results for S100-A4 protein, Figure 6c shows the results for S100-A6 protein, Figure 6d shows the results for S100-A7 protein, Figure 6e shows the results for S100-A8 protein, Figure 6f shows the results for S100-A9 protein, Figure 6g shows the results for S100-A10 protein, Figure 6h shows the results for S100-A13 protein, Figure 6i shows the results for S100-B protein, and Figure 6j shows the results for vimentin. (F1-F10 represent MPNST site samples from patients, and F11-F20 represent benign NF site samples from patients. Specifically, F1 and F2 are MPNST site samples from patient 1, F3 and F4 are MPNST site samples from patient 2, F5 and F6 are MPNST site samples from patient 3, F7 and F8 are MPNST site samples from patient 4, F9 and F10 are MPNST site samples from patient 5, F11 and F12 are benign NF site samples from patient 1, F13 and F14 are benign NF site samples from patient 2, F15 and F16 are benign NF site samples from patient 3, F17 and F18 are benign NF site samples from patient 4, and F19 and F20 are benign NF site samples from patient 5. (See Table 3)) [Figure 7] Figure 7 is a graph showing the results of an investigation into the expression patterns of endosialin in benign neurofibroma (NF) and malignant peripheral nerve schwannoma (MPNST) tissues. [Figure 8] Figure 8 is a graph showing the results of investigating the expression patterns of SPARC-like protein 1 in benign neurofibroma (NF) and malignant peripheral nerve schwannoma (MPNST) tissues. [Figure 9] Figure 9 is a graph showing the results of an investigation into the expression patterns of neutrophil collagenase in benign neurofibroma (NF) and malignant peripheral nerve schwannoma (MPNST) tissues. [Figures 10a-10c]Figures 10a to 10c show the results of hierarchical cluster analysis of the proteome with altered expression in benign neurofibroma (NF) and malignant peripheral nerve schwannoma (MPNST) tissues. Figure 10a visualizes the overall results, Figure 10b shows clusters of proteins with increased expression in MPNST, and Figure 10c shows clusters of proteins with decreased expression in MPNST. [Figure 11] Figure 11 shows the results of gene ontology analysis of malignant peripheral nerve schwannoma (MPNST) specific proteins. [Figure 12] Figure 12 shows the results of IPA (ingenuity pathway analysis) for malignant peripheral nerve schwannoma (MPNST) specific proteins. [Figure 13-21] Figures 13 to 21 are graphs comparing the abundance of kinases that were specifically expressed or had increased activity in malignant peripheral nerve schwannomas (MPNSTs) compared to benign neurofibromas (NFs). Figure 13 shows the results for CDK4, Figure 14 for MAP2K1, Figure 15 for PDK3, Figure 16 for STK10, Figure 17 for ALPK3, Figure 18 for PHKG1, Figure 19 for CDK5, Figure 20 for PNKP, and Figure 21 for PACSIN3 abundance. [Figure 22] Figure 22 shows the results of IHC (Immunohistochemistry) for the MMP8 protein (neutrophil collagenase) in benign neurofibroma (NF) and malignant peripheral nerve schwannoma (MPNST) tissues (A1 and B1 are NF tissues, and A2 and B2 are MPNST tissues). [Figure 23] Figure 23 shows the results of an ELISA (enzyme-linked immunosorbent assay) analysis of the difference in MMP8 protein (neutrophil collagenase) expression between patients with malignant peripheral nerve schwannoma (MPNST) and those who had tumor removal. [Figure 24]Figure 24 shows the results of immunochemistry (IHC) for the ILK signaling pathway, actin-cytoskeleton signaling pathway, and kinases that were specifically increased in expression or activity in malignant peripheral nerve schwannomas (MPNSTs) compared to benign neurofibromas (NFs) (A1-J1 are NF tissues, A2-J2 are MPNST tissues). [Modes for carrying out the invention]

[0044] The present invention relates to multiple biomarkers for the differential diagnosis of the progression from neurofibromatosis type I to malignant peripheral nerve sheath tumor (MPNST), and to a screening method for MPNST therapeutic agents that target signaling pathways or kinases whose expression is increased specifically in MPNST. The inventors have diligently researched to discover MPNST-specific diagnostic markers and novel therapeutic targets, and as a result have discovered multiple proteins and signaling pathways disclosed herein, leading to the completion of the present invention.

[0045] The present invention will be described in detail below.

[0046] The present invention relates to a composition for the differential diagnosis of malignant peripheral nerve sheath tumor (MPNST), comprising as an active ingredient a formulation for measuring the mRNA expression level of at least one protein selected from the group consisting of proteins listed in Table 1 below.

[0047] [Table 1]

[0048] In one embodiment of the present invention, through proteomic analysis using malignant peripheral nerve schwannomas (MPNSTs) and benign neurofibromas (NFs) and their tissues, MPNST-specific proteins were identified, and it was shown that three proteins detected from blood and urine—endosialin, SPARC-like protein 1, and neutrophil collagenase—can be selected and used as biomarkers for MPNST (see Examples 2-4 and 6).

[0049] The endosialin may be encoded by the CD248 gene and consist of the amino acid sequence of SEQ ID NO: 1, and may include an isomorphic protein in which amino acids 1 to 324 are missing from the amino acid sequence of SEQ ID NO: 1. The SPARC-like protein 1 may be encoded by the SPARCL1 gene and consist of the amino acid sequence of SEQ ID NO: 2, and may include an isomorphic protein in which amino acids 1 to 125 are missing from the amino acid sequence of SEQ ID NO: 2. Furthermore, the neutrophil collagenase may be encoded by the MMP8 gene and consist of the amino acid sequence of SEQ ID NO: 3.

[0050] As used herein, the term "protein" is used interchangeably with "polypeptide" or "peptide" and refers to a polymer of amino acid residues, as commonly found in natural proteins, for example. "mRNA" refers to RNA that transmits genetic information (gene-specific base sequences) from a specific gene to ribosomes, which identify the amino acid sequence during the process of protein synthesis.

[0051] As used herein, the term "expression" means the production of a protein or nucleic acid in a cell.

[0052] As used herein, "differential diagnosis" refers to distinguishing a particular disease or condition from others that present similar symptoms. Differential diagnostic methods are systematic diagnostic methods used to identify the existence of multiple possible conditions. Such methods are essentially elimination or information-gathering processes that reduce the probability of candidate conditions to a negligible degree. The term is also used to refer to determining whether or not there has been a transition from benign neurofibromatosis to malignant peripheral nerve sheath.

[0053] In the present invention, the composition may be used to distinguish and diagnose neurofibromatosis type 1 (NF-1) from malignant peripheral nerve schwannoma (MPNST).

[0054] As used herein, "neurofibromatosis type I" or "benign neurofibroma" refers to a genetic disorder that presents with various clinical symptoms in the skin, skeletal system, nervous system, etc., and was first reported in 1882 by Fredrich von Recklinghaus. Neurofibromatosis type I is characterized by café-au-lait spots on the skin, axillary freckling, inguinal freckling, multiple neurofibromas, Lisch nodules (small, tinted hamartomas in the iris), optic glioma, and osteoplasia. Other possible symptoms include short stature, scoliosis, and learning disabilities. Most tumors in patients with neurofibromatosis type 1 are benign, but in rare cases they can progress to malignancy and can occur in any part of the body with nerves. Studies have shown that 67% of cases are discovered before the age of one, 25-90% are accompanied by a characteristic skin lesion called a café-au-lait spot, and up to 16% become malignant.

[0055] As used herein, the term "malignant peripheral nerve sheath tumor (MPNST)" is used interchangeably with "malignant peripheral nerve sheath tumor." It generally occurs in the limbs and head and neck, and rarely in the abdominal cavity, pelvis, and genitourinary system. It is mainly associated with neurofibromatosis type 1, and it is known that approximately 10% of neurofibromatosis patients will develop a malignant peripheral nerve sheath tumor during their lifetime. It is one of the rare diseases with a very poor prognosis and high malignancy.

[0056] If the composition of the present invention is for measuring the expression level of a protein, the formulation for measuring the expression level of the protein may be, but is not limited to, an antibody or aptamer specific to the protein.

[0057] As used herein, the term “antibody” means a specific protein molecule directed to an antigenic site. For the purposes of the present invention, an antibody means an antibody that specifically binds to a marker protein, and includes polyclonal antibodies, monoclonal antibodies, and recombinant antibodies. Furthermore, any part of an antibody that has antigen-antibody binding properties is also included in the antibodies of the present invention, and includes all types of immunoglobulin antibodies that specifically bind to the aforementioned protein. For example, this includes not only complete antibodies having two full-length light chains and two full-length heavy chains, but also functional fragments of antibody molecules, i.e., Fab, F(ab'), F(ab')2, and Fv, which have antigen-binding function. In addition, the antibodies of the present invention also include special antibodies such as humanized antibodies and chimeric antibodies, as well as recombinant antibodies, as long as they can specifically bind to the aforementioned protein.

[0058] As used herein, the term "aptamer" refers to a single-stranded nucleic acid (DNA, RNA, or modified nucleic acid) that can specifically bind to an analyte to be detected in a sample and has a stable tertiary structure of its own, thereby specifically confirming the presence of a target protein in the sample. The production of aptamers may be carried out by determining and synthesizing an oligonucleotide sequence that has a selective and high binding affinity to the target protein to be detected using a general aptamer production method, and then modifying the 5' or 3' end of the oligonucleotide to -SH, -COOH, -OH, or NH2 so that it can be bound to the functional group of the aptamer tip, but is not limited to this method.

[0059] The composition of the present invention, comprising antibodies specific to the proteins listed in Table 1, may further comprise formulations necessary for a method of sensing known proteins, and the composition can be used without limitation to measure the expression level of a protein in a subject (in the present invention, a biological sample obtained from a subject).

[0060] Furthermore, if the composition of the present invention is for measuring the expression level of mRNA, the formulation for measuring the expression level of mRNA may be, but is not limited to, a primer set or probe that specifically binds to the mRNA.

[0061] As used herein, the term "primer" refers to a short nucleic acid sequence having a short free three-terminal hydroxyl group that can form base pairs with a complementary template and serve as a starting point for template strand copying. A primer can initiate DNA synthesis in the presence of reagents for polymerization (i.e., DNA polymerase or reverse transcriptase) and four different nucleoside triphodes at a suitable buffer solution and temperature.

[0062] As used herein, the term "probe" refers to a nucleic acid fragment, such as RNA or DNA, that can specifically bind to mRNA and corresponds to a short fragment of a few nucleotides or a long fragment of several hundred nucleotides, and is labeled to confirm the presence or absence of a specific mRNA. Probes may be prepared in the form of oligonucleotide probes, single-stranded DNA probes, double-stranded DNA probes, RNA probes, etc. The selection of appropriate probes and hybridization conditions may be modified based on those known in the art.

[0063] The "primers" or "probes" may be chemically synthesized using phosphoramidite solid support synthesis methods or other known methods. Furthermore, the primers or probes can be modified in various ways by methods known in the art, as long as this does not hinder hybridization with mRNA. Examples of such modifications include methylation, capping, substitution of one or more congeners of native nucleotides, and modification of nucleotides, such as uncharged conjugates (e.g., methylphosphonates, phosphotryesters, phosphoramidates, carbamates, etc.) or charged conjugates (e.g., phosphorothioates, phosphorodithioates, etc.), and the attachment of fluorescent or enzymatic labeling materials.

[0064] The composition of the present invention, comprising a primer set or probe specific to the mRNA of the proteins listed in Table 1, may further contain agents necessary for known RNA sensing methods. By using known RNA sensing methods with the composition of the present invention without limitation, the level of mRNA of the protein marker can be measured in a subject.

[0065] Furthermore, in the present invention, since the nucleic acid sequences of the genes encoding the mRNA of the protein are registered in the gene bank, those skilled in the art can design antisense oligonucleotides, primer pairs, or probes that specifically amplify specific regions of these genes based on the sequences.

[0066] The antisense oligonucleotides may be chemically synthesized using the phosphoramidite solid support method or other known methods, and include all functional equivalents. The same provisions regarding the "primers" or "probes" described above may also apply.

[0067] The term “functional equivalent” as used herein refers to all nucleotide and nucleic acid sequences of altered mutant sequences, such as, for example, one or more substitutions, deletions or additions from a reference sequence, or net effects that do not result in various functional dissimilarity between the reference sequence and the subject sequence. Typically, sequences that are substantial equivalents are about 35% (i.e., the number of substitutions, additions, and deletions of each residue in a substantially equivalent sequence is about 0.35 or less when divided by the total remaining number in the substantially equivalent sequence compared to the corresponding reference sequence) and are diverse from those enumerated herein. Such sequences have 65% sequence identity with the enumerated sequences. Substantially equivalent mutant, amino acid sequences according to the present invention preferably have at least 80% sequence identity, more preferably at least 90% sequence identity, with the enumerated amino acid sequences. Nucleotide sequences of the present invention that are substantial equivalents may have a lower percentage of sequence identity, for example, when considering redundancy or degeneracy of the genetic code. Preferably, the nucleotide sequences should have at least about 65% identity, more preferably at least about 75% identity, and most preferably about 95% identity. For the purposes of the present invention, sequences having substantially equivalent biological activity and substantially equivalent synthetic features are treated as substantial equivalents. In determining equivalents, truncation of mature sequences (e.g., due to mutations producing spurious stop codons) must be ignored.

[0068] In another aspect of the present invention, the present invention may provide a kit for the differential diagnosis of malignant peripheral nerve schwannoma comprising the composition according to the present invention.

[0069] In the present invention, the kit may include instructions indicating that if the expression of at least one protein or its mRNA selected from the group consisting of endosialin, SPARC-like protein 1, and neutrophil collagenase is detected, or if their expression levels are higher than those of a benign neurofibroma sample, it is determined to be a malignant peripheral schwannoma.

[0070] The kit of the present invention may include not only an antibody that recognizes a target protein as a marker or a primer set that recognizes mRNA as a marker, and a probe, but also one or more other component compositions, solutions, or apparatus suitable for the analytical method.

[0071] In specific embodiments, the diagnostic kit may be a diagnostic kit characterized by containing essential elements necessary for performing a reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit includes a set of primers specific to each marker gene. The primers are nucleotides having sequences specific to the nucleic acid sequence of each marker gene, and are about 7 bp to 50 bp long, more preferably about 10 bp to 30 bp long. It may also include primers specific to the nucleic acid sequence of a control gene. Other reverse transcription polymerase reaction kits may include a test tube or other suitable container, reaction buffer (pH and magnesium concentration vary), deoxyribonucleotides (dNTPs), enzymes such as Taq polymerase and reverse transcriptase, DNAse, the RNase inhibitor DEPC-water, sterile water, etc.

[0072] Another specific embodiment may be a diagnostic kit characterized by containing essential elements necessary for DNA chipping. The DNA chip kit may include a substrate to which cDNA or oligonucleotides corresponding to a gene or fragment of a gene are attached, and reagents, formulations, enzymes, etc., for preparing a fluorescently labeled probe. The substrate may also contain cDNA or oligonucleotides corresponding to a control group gene or fragment of a control group.

[0073] In yet another aspect of the present invention, the present invention may provide a method for providing information necessary for the differential diagnosis of malignant peripheral nerve schwannoma, comprising the step of measuring the mRNA expression level of at least one protein selected from the group consisting of endosialin, SPARC-like protein 1, and neutrophil collagenase in a biological sample isolated from a subject.

[0074] In yet another aspect of the present invention, the present invention provides a method for the differential diagnosis of malignant peripheral nerve schwannoma, comprising the step of measuring the mRNA expression level of at least one protein selected from the group consisting of endosialin, SPARC-like protein 1, and neutrophil collagenase in a biological sample isolated from a subject.

[0075] The differential diagnostic method for malignant peripheral nerve sheath tumors according to the present invention can rapidly and conveniently diagnose the progression from neurofibromatosis type I to malignant peripheral nerve sheath tumor (MPNST) not only through lesional tissue using multiple biomarkers but also through bodily fluid samples (for example, urine or plasma), thereby enabling immediate response to malignant peripheral nerve sheath tumors.

[0076] Therefore, the differential diagnostic method for malignant peripheral nerve schwannomas according to the present invention may be effectively used in the treatment of neurofibromatosis type 1 and other conditions.

[0077] In the present invention, the method may further include the step of diagnosing a malignant peripheral schwannoma if the expression of at least one protein or its mRNA selected from the group consisting of endosialin, SPARC-like protein 1, and neutrophil collagenase is detected, or if the expression levels thereof are higher than the expression levels in biological samples isolated from patients with neurofibromatosis type 1.

[0078] In yet another aspect of the present invention, the present invention may provide a method for treating malignant peripheral nerve schwannoma, comprising the following steps:

[0079] (a) A step of measuring the mRNA expression level of at least one protein selected from the group consisting of endosialin, SPARC-like protein 1, and neutrophil collagenase in a biological sample isolated from the subject.

[0080] (b) If the expression of at least one protein or its mRNA selected from the group consisting of endosialin, SPARC-like protein 1, and neutrophil collagenase is detected, or if their expression levels are higher than those in biological samples isolated from patients with neurofibromatosis type 1, the step of diagnosing malignant peripheral schwannoma, and

[0081] (c) The stage of treating the malignant peripheral nerve schwannoma diagnosed in the above (b) stage.

[0082] In the present invention, the step of treating the malignant peripheral nerve schwannoma may involve methods such as chemotherapy, radiotherapy, surgery, or biological therapy.

[0083] In this invention, chemotherapy means the act of using chemical substances to treat a particular disease and the entirety of the drugs used in that process.

[0084] In the present invention, the drugs include, for example, paclitaxel, doxorubicin, 5-fluorouracil, cisplatin, imatinib, carboplatin, oxaliplatin, tegafur, irinotecan, docetaxel, cyclophosphamide, cemcitabine, ifosfamide, and mitomycin C. C), vincristine, etoposide, methotrexate, topotecan, tamoxifen, vinorelbine, camptothecin, daunorubicin, chlorambucil, bryostatin-1, calicheamicin, mayatansine, levamisole, DNA recombinant interferon α-2a, mitoxantrone, nimustine, interferon α-2a ALFA-2A), doxifluridine, formestane, leuprolide acetate, megestrol acetate(acetic acid), carmofur, teniposide, bleomycin, carmustine, heptaplatin, exemestane, anastrozole, estramustine, capecitabine, goserelin acetate, polysaccharide potassium, medroxyprogesterone acetate, epirubicin, letrozole, pirarubicin, topotecan, altretamine, toremifene citrate The anticancer agent may be, but is not limited to, at least one selected from the group consisting of citrate, BCNU, taxotere, actinomycin D, and their synthetic analogues, and modified or equivalent substances.

[0085] In the present invention, the drug may be an antagonist for the protein listed in Table 2 below, for example, selumetinib, alvocidib, purvalanol, palbociclib, ribociclib, abemaciclib, fostamatinib, binimetinib, radicicol, AZD-8330, dihydrolipoic acid, which are inhibitors of the signal transduction pathway protein activity listed in Table 2. At least one selected from the group consisting of acid, alsterpaullone, hymenialdisine, indirubin-3'-monoxime, olomoucine, SU9516, and 6-phenyl[5H]pyrrolo[2,3-B]pyrazine may be present, but is not limited to.

[0086] In the present invention, the radiotherapy means exposing a patient to high-energy radiation, including but not limited to X-rays, gamma rays, and neutrons. Examples of such therapies include, but are not limited to, external phototherapy, internal radiation therapy, insertion radiation, proximal radiation therapy, and whole-body radiation therapy.

[0087] In the present invention, the surgical procedure includes all therapeutic or diagnostic procedures that involve the hand and instrumental methods of the hand on the body of an individual in order to obtain a healing, therapeutic, or diagnostic effect.

[0088] In the present invention, the biological therapy refers to a treatment method that directly or indirectly uses the human body's immune system using a biological preparation containing a substance derived from a living organism or a substance produced using a living organism. The biological preparation includes vaccines, allergens, antigens, hormones, cytokines, enzymes, blood and plasma, immune serum, monoclonal antibodies, fermented products, antitoxins, and laboratory diagnostic agents, whose potency and stability cannot be evaluated by physical and chemical tests alone.

[0089] In the present invention, the biological agent is, for example, adalimumab, alemtuzumab, bevacizumab, cetuximab, daratumumab, panitumumab, rituximab, trastuzumab, pertuzumab, ipilimumab. It may be at least one selected from the group consisting of mumab, nivolumab, pembrolizumab, atezolizumab, durvalumab, avelumab, tocilizumab, sarilumab, satralizumab, and siltuximab. However, it is not limited to these.

[0090] As used herein, "detection" includes measuring and confirming the presence or absence of a desired substance (marker protein in this invention), or measuring and confirming a change in the level of the desired substance (expression level). In the same context, measuring the expression level of the protein means measuring the presence or absence of expression (i.e., measuring the presence or absence of expression), or measuring the level of qualitative or quantitative change in the protein. The measurement may be carried out without limitation, including both qualitative (analytical) and quantitative methods. The types of qualitative and quantitative methods for measuring protein levels are well known in the art, and the experimental methods described herein are included therein. Specific protein level comparison methods for each method are well known in the art. Therefore, the detection of the target protein includes detecting the presence or absence of the proteins listed in Table 1, or confirming an increase (upward regulation) or decrease (downward regulation) in protein expression levels.

[0091] Furthermore, the term "differential diagnosis" in the foregoing sense includes, in addition to the meanings described above, determining the susceptibility of a subject to a particular disease or disorder, determining whether a subject currently has a particular disease or disorder, determining the prognosis of a subject suffering from a particular disease or disorder, or terametrics (for example, monitoring the state of an object to provide information on therapeutic efficacy).

[0092] In the present invention, the subject may be a patient suspected of having malignant peripheral nerve schwannoma, or a patient diagnosed with neurofibromatosis type 1.

[0093] In the present invention, the biological sample may be used without limitation as long as it is taken from a subject who is to be subjected to differential diagnosis of malignant peripheral nerve schwannoma. For example, it may include not only biological samples such as tissue, cells, blood, plasma, serum, saliva, nasal fluid, sputum, joint fluid, amniotic fluid, ascites, cervical secretions, vaginal secretions, urine, and cerebrospinal fluid obtained by biopsy, but also microneedle aspiration samples and microneedle lavage solutions. Preferably, the measurement may be performed using at least one biological liquid sample selected from the group consisting of blood, whole blood, plasma, urine, tissue, cells, organs, bone marrow, microneedle aspiration samples, microneedle lavage solutions, core needle biopsy samples, and saliva. The present invention has great technical significance in that it enables diagnosis not only through lesional tissue in which malignant peripheral nerve schwannoma has occurred, but also through bodily fluid samples (for example, urine or plasma), thereby increasing the efficacy, speed, convenience, and safety of the diagnosis.

[0094] The biological sample may be pretreated before use for detection or diagnosis. This may include, for example, homogenization, filtration, distillation, extraction, concentration, inactivation of interfering components, and addition of reagents. The sample may be prepared to increase the sensitivity of the protein marker, but for example, serum samples obtained from patients may be pretreated using methods such as anion exchange chromatography, affinity chromatography, size exclusion chromatography, liquid chromatography, sequential extraction, or gel electrophoresis.

[0095] The measurement of the protein expression level is not particularly limited as long as it is performed using a protein expression measurement method known in the industry, but may be at least one selected from the group consisting of, for example, Western blotting, ELISA, radioimmunoanalysis, radioimmunodiffusion, Ouchterlony immunodiffusion, Rocket immunoelectrophoresis, immunostaining, immunoprecipitation analysis, complement fixation analysis, mass spectrometry, FACS, and protein chips.

[0096] The mRNA expression level measurement may be at least one selected from the group consisting of PCR, RNase protection analysis, northern blotting, southern blotting, in situ cross-pollination, and DNA chip, using gene expression measurement methods known in the industry, but is not limited thereto.

[0097] As used herein, the term “analysis” may preferably mean “measurement,” and such qualitative analysis may mean measuring and confirming the presence or absence of a desired substance, and such quantitative analysis may mean measuring and confirming a change in the level of presence (expression level) or amount of a desired substance. In the present invention, analysis or measurement may be carried out without limitation including both qualitative and quantitative methods, and preferably quantitative measurement may be performed.

[0098] On the other hand, the mechanisms of MPNST development in NF-1 have been known to involve the complete disappearance of NF-1 within tumors, excessive activation of the RAS-MAPT signaling pathway, loss of the CDKN2A gene, loss of the PRC2 (polycomb repressive complex 2) constituent genes SUZ12 and EED, loss of the tumor suppressor gene TP53, abnormal enhancement of the PI3K-AKT-mTOR signaling pathway, and abnormal activation of EGFR (epidermal growth factor receptor), PDGF-R (platelet-derived growth factor receptor), and VEGFR (vascular endothelial growth factor receptor). Based on this, various clinical studies have been conducted on MPNST, but none have shown clear therapeutic effects. Therefore, there was a need to explore new therapeutic targets and candidate substances using a novel approach.

[0099] In response to such demands, the present invention identifies four signaling pathways specifically enhanced in malignant peripheral nerve schwannomas (MPNSTs)—the ILK signaling pathway, the actin-cytoskeleton signaling pathway, the RhoGDI signaling pathway, and the TGF-β signaling pathway—and ten kinases specifically expressed or with increased activity in MPNSTs—the kinases expressed by the CDK4, MAP2K1, PDK3, STK10, ALPK3, PHKG1, CDK5, PNKP, CFL1, and PACSIN3 genes—and confirms that these can be utilized as novel therapeutic targets for MPNSTs (see Examples 5 and 7).

[0100] Therefore, in yet another aspect of the present invention, the present invention may provide a pharmaceutical composition for the prevention or treatment of malignant peripheral nerve schwannoma, comprising as an active ingredient an inhibitor of the activity of at least one signaling pathway protein selected from the group consisting of the TGF-β signaling pathway, the ILK signaling pathway, the actin-cytoskeleton signaling pathway, and the RhoGDI signaling pathway, or an inhibitor of the expression of a gene encoding said protein.

[0101] In yet another aspect of the present invention, the present invention may provide a health functional food composition for the prevention or improvement of malignant peripheral nerve schwannoma, comprising as an active ingredient an inhibitor of the activity of at least one signaling pathway protein selected from the group consisting of the TGF-β signaling pathway, the ILK signaling pathway, the actin-cytoskeleton signaling pathway, and the RhoGDI signaling pathway, or an expression inhibitor of the gene encoding said protein.

[0102] The aforementioned "TGF-β signaling pathway proteins" include all proteins directly or indirectly involved in TGF-β signaling. Examples include SMAD1, SMAD2, SMAD3, SMAD5, SMAD9, Thrombospondin-1, Rock, Cdc42 / RAC, PAK, LIMK, PI3K, Ras, Raf, MEK1 / 2, ERK1 / 2, JNK, P38, Cofilin, mTOR, and AKT.

[0103] The aforementioned "ILK signaling pathway (Integrin-linked kinase signaling) proteins" include all proteins directly or indirectly involved in ILK signaling. Examples include mTOR, AKT, VEGF, NF-κGSK3β, SNAIL, AP-1, MMP9, JAK2, Cdc42 / RAC, SNAI1, JNK, PI3K, AP-1, C-Jun, HIF1α, COX2, and β-Catenin.

[0104] The aforementioned actin-cytoskeleton signaling proteins include all proteins directly or indirectly involved in actin-cytoskeleton signaling. Examples include PI3K, CaM, PKA, Rho, ROCK, Ras, Rac, Paxillin, FAK, Cofilin, Cortactin, WAVE, PIX, TIAM1, Vav, PAK1, GEF, VASP, MEKK, MEK, MLCK, MLC, LIMK, ADF, CapZ, Hsp90, Gelsolin, ADF, Filamin, Profilin, RhoGEF, Myosin, ACTN, Vinculin, ACTN, PFN, MEK1 / 2, and ERK1 / 2.

[0105] The aforementioned "RhoGDI signaling pathway (Rho GDP-dissociation inhibitor signaling) proteins" include all proteins directly or indirectly involved in RhoGDI signaling. Examples include MKK4 / 7, JNK, C-Jun, PARP, ROCK1, ROCK2, Cdc42, eNOS, Tau, MLCP, MLC, CRMP2, Adducin, LIMK, PKN, BORGs, Citron, WASP, MRCK, PI3K, PAK, PAR6, and PAR3.

[0106] In the present invention, the signal transduction pathway protein may be a kinase, and the kinase may be at least one kinase selected from the group consisting of the proteins listed in Table 2 below, but is not limited thereto.

[0107] In the present invention, the kinases listed in Table 2 below may belong to the ILK signaling pathway ("CDK4", "STLK10", and "PHKG1"), the actin-cytoskeleton signaling pathway and the RhoGDI signaling pathway ("CDK5"), the actin-cytoskeleton signaling pathway and the TGF-β signaling pathway ("MAP2K1" and "CFL1"), the heart development pathway ("ALPK3"), the DNA damage and repair pathway ("PNKP"), the pyruvate metabolism and citric acid (TCA) cycle pathway ("PDK3"), and the actin-cytoskeleton signaling pathway ("PACSIN3"), but are not limited to these.

[0108] [Table 2]

[0109] In this invention, it has been confirmed that the proteins listed in Table 2 (including p-MEK) are specifically regulated upward in MPNST. Therefore, substances that can inhibit these proteins, such as antagonists, may be used as MPNST therapeutic agents.

[0110] In the present invention, the protein activity inhibitor may be at least one selected from the group consisting of peptides, antibodies, aptamers, and compounds that specifically bind to the protein, but is not limited thereto.

[0111] The definitions of "antibody" and "aptamer" used in this specification are as stated above.

[0112] In the present invention, the inhibitors of the activity of the signaling pathway proteins are selumetinib, alvocidib, purvalanol, palbociclib, ribociclib, abemaciclib, fostamatinib, binimetinib, radicicol, AZD-8330, and dihydrolipoic acid. At least one selected from the group consisting of acid, alsterpaullone, hymenialdisine, indirubin-3'-monoxime, olomoucine, SU9516, and 6-phenyl[5H]pyrrolo[2,3-B]pyrazine may be present, but is not limited to.

[0113] In the present invention, the gene expression inhibitor may be at least one selected from the group consisting of miRNA, siRNA, shRNA, ribozyme, DNAzyme, PNA (peptide nucleic acid), and antisense oligonucleotide that bind complementarily to the mRNA of the gene, but is not limited thereto.

[0114] As used herein, the term “antisense oligonucleotide” encompasses nucleic acid-based molecules that have a sequence complementary to the target mRNA, particularly the mRNA seed sequence, and that can form a duplex with the mRNA. Therefore, the term “antisense oligonucleotide” may also be referred to herein as “complementary nucleic acid-based inhibitor.”

[0115] In this specification, the term “complementary” as used in reference to antisense oligonucleotides means that the antisense oligonucleotide is sufficiently complementary to selectively hybridize with the mRNA target under given hybridization or annealing conditions, preferably physiological conditions, and encompasses both substantially complementary and perfectly complementary, preferably perfectly complementary.

[0116] In the present invention, antisense oligonucleotides include a variety of molecules. Antisense oligonucleotides are DNA or RNA molecules, and more preferably RNA molecules. Selectively, antisense oligonucleotides used in the present invention are ribonucleotides (RNA), deoxyribonucleotides (DNA), 2'-O-modified oligonucleotides, phosphorothioate-backbone deoxyribonucleotides, PNA (peptide nucleic acid), or LNA (locked nucleic acid). 2'-O-modified oligonucleotides may be, for example, 2'-O-alkyl oligonucleotides, 2'-O-C1-3 alkyl oligonucleotides, or 2'-O-C1-3 methyl oligonucleotides.

[0117] As used herein, the terms “miRNA, siRNA, and shRNA” refer to nucleic acid molecules that bind primarily to mRNA transcribed from a target gene to mediate RNA interference or gene silencing, thereby inhibiting the detoxification of said mRNA. Because these miRNAs, siRNAs, and shRNAs can suppress the expression of target genes at a detoxification level, they may be used in efficient gene knockdown methods or gene therapies.

[0118] As used herein, the term "ribozyme" refers to a substance that can suppress the protein expression of a target gene by recognizing a specific nucleotide sequence within a target RNA molecule and cleaving it site-specifically.

[0119] As used herein, “PNA” means a nucleic acid mimetic, e.g., a DNA mimetic, where the deoxyribose phosphate backbone is replaced with a pseudopeptide backbone, but retaining the original four nucleobases. The neutral backbone of PNA is known to provide DNA and RNA-specific hybrids under low ionic strength conditions and may be used as an antisense or antigen preparation for sequence-specific regulation of gene expression by inducing transcriptional or translational repression or inhibiting replication.

[0120] On the other hand, the content of the signaling pathway protein activity inhibitor or the gene expression inhibitor encoding the protein in the composition of the present invention can be appropriately adjusted according to the symptoms of the disease, the progression of the symptoms, the patient's condition, etc. For example, it may be 0.0001 to 99.9% by weight or 0.001 to 50% by weight based on the total weight of the composition, but is not limited thereto. The content ratio is a value based on the dry weight after removing the solvent.

[0121] The pharmaceutical composition according to the present invention may further comprise suitable carriers, excipients, and diluents commonly used in the manufacture of pharmaceutical compositions. The excipient may be at least one selected from the group consisting of, for example, diluents, binders, disintegrants, lubricants, adsorbents, humectants, film coating substances, and controlled-release additives.

[0122] The pharmaceutical compositions according to the present invention may be used in the form of external preparations such as powders, granules, sustained-release granules, enteric-coated granules, liquids, eye drops, elsilics, emulsions, suspensions, alcoholic preparations, lozenges, aromatic preparations, limonades, tablets, sustained-release tablets, enteric-coated tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric-coated capsules, pills, tinctures, softened extracts, dried extracts, liquid extracts, injections, capsules, perfusion solutions, ointments, lotions, pastes, sprays, inhalants, patches, sterile injections, or aerosols, respectively, by conventional methods. The external preparations may also have dosage forms such as creams, gels, patches, sprays, ointments, ointments, lotions, liniments, pastes, or cataplasms.

[0123] Carriers, excipients, and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, amorphous cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0124] When formulating, the product is typically prepared using fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, or other diluents or excipients.

[0125] Excipients for tablets, powders, granules, capsules, pills, and lozenges according to the present invention include corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, monocalcium phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropyl methylcellulose (HPMC) 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, primozel, gelatin, gum arabic, ethanol, agar powder, cellulose phthalate acetate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, and sodium casein. Binders such as glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, refined shellac, starch paste, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, and polyvinylpyrrolidone may be used. Hydroxypropyl methylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, anhydrous silicic acid, 1-hydroxypropylcellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, and guar gum may be used.gum), baking soda, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, amylopectin, pectin, sodium polyphosphate, ethylcellulose, sucrose, magnesium aluminum silicate, D-sorbitol solution, disintegrants such as hard anhydrous silicic acid, calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium, kaolin, petrolatum, sodium stearate, cocoa butter, sodium salicylate, magnesium salicylate, polyethylene glycol (PEG) 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil (Lubri). Lubricants such as wax, aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, anhydrous silicic acid, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and hard anhydrous silicic acid may be used.

[0126] Additives to the liquid formulation according to the present invention may include water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearate, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethylcellulose, sodium carboxymethylcellulose, and the like.

[0127] The syrup according to the present invention may contain a solution of sucrose, other sugars, or sweeteners, and may also contain, if necessary, fragrances, colorants, preservatives, stabilizers, suspending agents, emulsifiers, viscosity enhancers, etc.

[0128] Purified water may be used in the emulsion of the present invention, and emulsifiers, preservatives, stabilizers, fragrances, etc. may be used as needed.

[0129] The suspending agent according to the present invention may contain suspending agents such as acacia, tragacantha, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropyl methylcellulose (HPMC), HPMC1828, HPMC2906, HPMC2910, and, if necessary, surfactants, preservatives, stabilizers, colorants, and fragrances may be used.

[0130] The injectable preparation according to the present invention includes distilled water for injection, 0.9% sodium chloride injection solution, Ringer's injection solution, dextrose injection solution, dextrose + sodium chloride injection solution, PEG (PEG), lactating Ringer's injection solution, solvents such as ethanol, propylene glycol, non-volatile oils - sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate, solubilizers such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidinedione, propylene glycol, twins, dijontinamide, hexamine, and dimethylacetamide, weak acids and their salts (acetic acid and sodium acetate), and weak bases and their salts (ammonia and ammonium acetate). It may also contain buffering agents such as organic compounds, proteins, albumin, peptone, and gums; isotonic agents such as sodium chloride; stabilizers such as sodium bisulfite (NaHSO3), carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen gas (N2), and ethylenediaminetetraacetic acid; sulfurizing agents such as 0.1% sodium bisulfide, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate, and sodium acetone bisulfite; analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; and suspending agents such as sodium CMC, sodium alginate, Twin 80, and aluminum monostearate.

[0131] The suppositories according to the present invention include cocoa butter, lanolin, vitepsol, polyethylene glycol, glycerol gelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, coconut oil, cocoa butter + cholesterol, lecithin, lanette wax, glycerol monostearate, twin or span, Imhausen, monolen (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego-G, and Cebes Pharma 16. 16) Hexalide Base 95, Cotomar, Hydrocoat SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydrocoat 25, Hydrocoat 711, Idropostal, Massa Estralium Bases such as estrarium (A, AS, B, C, D, E, I, T), Masa-MF, Maspol, Maspol-15, Neospostal-en, Paramount-B, Sposyl (OSI, OSIX, A, B, C, D, H, L), Suppository base type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Spostal (N, Es), Wecobi (W, R, S, M, Fs), and Tezestr triglyceride base (TG-95, MA, 57) may be used.

[0132] Orally administered solid dosage forms include tablets, pills, powders, granules, and capsules. Such solid dosage forms are prepared by mixing the extract with at least one excipient, such as starch, calcium carbonate, sucrose, or lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stylate talc are also used.

[0133] Oral liquid formulations include suspensions, liquid preparations, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as humectants, sweeteners, fragrances, and preservatives may be included. Parenteral formulations include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspension solvents may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

[0134] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, “pharmaceutically effective amount” means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level may be determined by factors including the type and severity of the patient’s disease, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and elimination ratio, the duration of treatment, drugs used concurrently, and other factors well known in the medical field.

[0135] The pharmaceutical compositions according to the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered as a single or multiple agent. Taking all of the above factors into consideration, it is important to administer an amount that provides the greatest effect with the minimum amount without side effects, which may be readily determined by a person of the skill in the art to which the present invention belongs.

[0136] The pharmaceutical composition of the present invention may be administered to an individual by various routes. All methods of administration are predictable, but may include, for example, oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, perispinal space (intradural) injection, sublingual administration, buccal administration, rectal insertion, vaginal insertion, ocular administration, ear administration, nasal administration, inhalation, spraying through the mouth or nose, skin administration, transdermal administration, etc.

[0137] The pharmaceutical composition of the present invention is determined by the type of drug that is the active ingredient, along with various relevant factors such as the disease to be treated, the route of administration, the patient's age, sex, weight, and the severity of the disease.

[0138] In this invention, "individual" means a subject requiring treatment for a disease, and more specifically, a mammal such as a human or non-human primate, mouse, rat, dog, cat, horse, or cow.

[0139] In the present invention, "administration" means providing a predetermined composition of the present invention to an individual by any appropriate method.

[0140] In the present invention, "prevention" means all actions that suppress or delay the onset of a desired disease, and "treatment" means all actions that improve or favorably alter the symptoms of a desired disease and the metabolic disorder caused thereby by administering a pharmaceutical composition according to the present invention.

[0141] As used herein, the term "health functional food" refers to a food manufactured and processed using raw materials or components that have beneficial functional properties for the human body, as defined in Act No. 6727 on Health Functional Foods. It means that the food is consumed for the purpose of regulating nutrients in relation to the structure and function of the human body, or for obtaining beneficial effects for health purposes such as physiological effects. The health functional food of the present invention may contain ordinary food additives, and unless otherwise specified, the suitability of the food additive shall be determined according to the standards and criteria for the item in question, such as the general provisions and general test methods of the Food Additives Code approved by the Ministry of Food and Drug Safety.

[0142] In yet another aspect of the present invention, the present invention may provide a method for screening for substances for the prevention, improvement, or treatment of malignant peripheral nerve schwannomas, comprising the steps of: (a) contacting isolated cells or tissues with a candidate substance; (b) measuring the activity of at least one signaling pathway protein selected from the group consisting of the TGF-β signaling pathway, the ILK signaling pathway, the actin-cytoskeleton signaling pathway, and the RhoGDI signaling pathway, or the expression level of a gene encoding the protein, in the cells or tissues; and (c) determining that the candidate substance is a substance for the prevention, improvement, or treatment of malignant peripheral nerve schwannomas if the activity of the protein or the expression level of the gene encoding the protein decreases in the cells or tissues that have been in contact with the candidate substance compared to the level before contact with the candidate substance.

[0143] In the present invention, the candidate substance may include, but is not limited to, low molecular weight compounds, high molecular weight compounds, microbial culture solutions or extracts, natural product extracts, nucleic acids, proteins, sugars, and lipids. It may also include any substance that can reduce the activity of at least one signaling pathway protein selected from the group consisting of the TGF-β signaling pathway, the ILK signaling pathway, the actin-cytoskeleton signaling pathway, and the RhoGDI signaling pathway, or the expression level of the gene encoding said protein.

[0144] The nucleic acid may be selected from the group consisting of microRNA, siRNA, shRNA, antisense RNA, aptamer, LNA (locked nucleic acid), PNA (peptide nucleic acid), and morpholino, but is not limited thereto.

[0145] In the present invention, the signal transduction pathway protein may be a kinase, and the kinase may be, but is not limited to, a kinase encoded by at least one gene selected from the group consisting of CDK4, STK10, CDK5, MAP2K1, ALPK3, PNKP, PDK3, PHKG1, CFL1, and PACSIN3.

[0146] In the present invention, the isolated cells or tissue may be, but is not limited to, nerve cells or nerve tissue.

[0147] The terminology used in this invention has been selected as widely used and general terms as possible, taking into consideration the function of the invention; however, this may change depending on the intentions of the articulators, case law, and the emergence of new technologies. In some cases, the applicant has arbitrarily selected terms, in which case their meaning will be described in detail in the description of the relevant invention. Therefore, the terminology used in this invention should not be merely names of terms, but should be defined based on the meaning of the term and the overall content of the invention.

[0148] Throughout the specification of this invention, when a part of it "includes" a certain component, this means that, unless otherwise stated, it may include other components rather than excluding them. Terms of degree used throughout the specification of this invention, such as "about," "substantially," etc., are used to mean the numerical value or a value close to the numerical value when manufacturing and material tolerances specific to the meaning mentioned are presented, and are used to prevent unscrupulous infringers from unfairly exploiting disclosures that mention precise and absolute numerical values ​​to aid in understanding the invention.

[0149] Throughout the specification of the present invention, the term “these combinations” as used in the Marcoush expression means one or more mixtures or combinations selected from the group of components described in the Marcoush expression, and means including one or more selected from the group of components. [Examples]

[0150] The following are preferred embodiments to aid in understanding the present invention. However, the following embodiments are provided to facilitate understanding of the present invention and do not limit the scope of the present invention.

[0151] [Examples] Example 1. Sample preparation and proteomic analysis 1.1. Sample Preparation Malignant peripheral nerve schwannoma (MPNST) tissue was obtained from five patients diagnosed with neurofibromatosis type 1, and benign neurofibroma tissue from five patients was used as a control group. Information on the MPNST patients is shown in Table 3 below.

[0152] [Table 3]

[0153] The obtained tissue samples were fixed with formalin and then embedded in paraffin. For proteome separation, the FFPE (formalin-fixed paraffin-embedded) tissue slides were separated with a razor blade and placed in tubes, as shown in the flowcharts in Figures 1a and 1b, and deparaffinized with 200 μl of heptane at 25°C for 1 hour. Next, to crush, extract, and homogenize the samples, the samples were lysed using an AFA (adaptive focused acoustics) ultrasonic grinder after adding 100 μl of 5% SDS and 50 mM Triethylammonium biocarbonate buffer. Subsequently, the samples were digested using Trypsin / LysC protease to produce peptides, and the proteome was analyzed using LC-MS.

[0154] 1.2. Proteomic Analysis Proteome analysis was performed using LC-MS. The mass spectrometry (MS) equipment used was the Q-Exactive Plus BioPharm version (ThermoFisher, USA), and the liquid chromatography (LC) equipment was the UltiMate. TM A 3000 RSLC nano System (ThermoFisher, USA) was used. Samples were injected in 5.0 μl increments using an automated sample injector, and analysis was performed using an ion trap mass spectrometer (ILC) linked to the NanoLC at a flow rate of 250 nl / min.

[0155] The LC column used was a fused silica capillary column C18 with an inner diameter of 75 μm and an outer diameter of 360 μm. The sample was analyzed for 200 minutes at a flow rate of 250 nl / min, and the concentration gradient was separated from 5% solution B to 50% solution B (80% acetonitrile, 0.1% formic acid, 5% DMSO) for 90 minutes.

[0156] The parameters for MS were as follows: MS1 resolution 70000, MS1 maximum fill time 20 msec, DDA (data dependent acquisition) method (top 10); MS2 resolution 17500, MS2 maximum fill time 100 msec, AGC (auto gain control) 1e6.

[0157] Example 2. Analysis of proteome data and identification of malignant peripheral nerve schwannoma (MPNST) specific proteins therefrom. Proteomic data analysis was performed on the malignant peripheral nerve schwannoma (MPNST) and benign neurofibroma (NF) tissue samples obtained in Example 1.

[0158] First, as shown in Figure 2, raw data was obtained using Proteome Discovery 2.2 software (Thermo, USA). Qualitative and quantitative information on proteins was extracted using a label-free quantification algorithm with the human proteome reference database provided by Uniprot. For qualitative analysis, the fixed modification condition for peptide search was cysteine ​​with carbamidomethylation, and the variable modification conditions were n-terminal acetylation and methionine oxidation. The initial mass error for precursor ions was 10 ppm, and the mass error for fragment ions was adjusted to 20 ppm. When mapping peptides to proteins, all unique peptides and razor peptides were used. For label-free quantification, the peak intensity of the unique peptide was used.

[0159] As a result, as shown in Figure 3, we confirmed that the distribution of protein quantification values ​​obtained from each sample was uniform.

[0160] Furthermore, principal component analysis (PCA) was performed using the proteome quantitative information of the aforementioned samples. As a result, as shown in Figure 4, it was confirmed that the two groups, MPNST and NF, could be distinguished by quantitative and qualitative differences in their proteomes. More specifically, referring to the right graph in Figure 4, 3,483 proteins were identified in benign NF tissue and 3,601 proteins were identified in MPNST tissue. Additionally, 141 proteins were not identified in benign NF tissue but were identified only in MPNST tissue, and vice versa, with 23 proteins being identified in the reverse case.

[0161] Example 3. Derivation of proteins whose expression was increased in malignant peripheral nerve schwannoma (MPNST). Through quantitative proteome analysis using volcano plot analysis, as shown in Figure 5a, we investigated proteins that were more than 10-fold increased in expression in MPNST compared to benign NF, and which were mainly expressed in extracellular exosomes (indicated by red boxes). This is advantageous because proteins excreted into exosomes can be detected in blood and urine, making it easier to collect samples for diagnosis.

[0162] As a result, 24 proteins, including CD248, DNAJB4, POTEE, SPARCL1, and TBC1D4, were selected, as shown on the right side of Figure 5a.

[0163] Furthermore, as shown in Figure 5b, proteins that degrade the extracellular matrix (proteinaceous extracellular matrix) were investigated (indicated by red boxes). This is because proteins expressed in the extracellular matrix can also be detected in blood and urine. Moreover, since matrix degradation is a phenomenon that occurs when cancer cells invade surrounding tissues, these proteins are related to the malignancy of the cells.

[0164] As a result, the CD248, SPARCL1, CILP2, MMP8, and SPON1 proteins were selected, as shown on the right side of Figure 5b.

[0165] Comparative Example 1. Investigation of S-100 protein expression patterns in malignant peripheral nerve schwannoma (MPNST) and benign neurofibroma (NF). We investigated the expression patterns of the S-100 protein, which has traditionally been used as an important pathological indicator for distinguishing between benign NF and MPNST.

[0166] When a total of nine S-100 isoforms were investigated, differences in expression patterns between benign NF and MPNST were observed depending on the individual patient, as shown in Figures 6a-6i. These results clearly demonstrate the limitations of the S-100 protein as a specific marker for distinguishing between benign NF and MPNST.

[0167] Furthermore, when we investigated vimentin as another indicator to distinguish between benign NF and MPNST, we found no difference in expression patterns between benign NF and MPNST.

[0168] In summary, the results suggest that the conventionally used markers S-100 and vimentin have clear limitations for clinical application as MPNST-specific biomarkers.

[0169] Example 4. Selection of diagnostic markers for malignant peripheral nerve schwannoma (MPNST) 4.1. Endocialin (CD248) Endosialin, a protein encoded by the CD248 gene, is a molecule involved in tumor angiogenesis. It is secreted into the extracellular matrix and is also found in exosomes.

[0170] As can be seen in Figure 7, endosialin was specifically found in all MPNST tissues obtained from five patients. In particular, since endosialin is found in exosomes, it is expected to be highly useful as an MPNST diagnostic marker because it can be detected in blood and urine samples.

[0171] 4.2. SPARC-like protein 1 (SPARCL1) SPARC-like protein 1, encoded by the SPARCL1 gene, is a protein that is primarily secreted outside the cell and binds to calcium, extracellular matrix, collagen, and other substances.

[0172] As shown in Figure 8, SPARC-like protein 1 was not detected in benign NF but was specifically expressed in MPNST. Since this protein is also found extracellularly and is present in exosomes, it is expected to be detectable in blood and urine samples, and therefore is likely to be widely used as a diagnostic marker for MPNST.

[0173] 4.3. Nutrofil collagenase (MMP8) Neutrophil collagenase, encoded by the MMP8 gene, is a protease that degrades the extracellular matrix and is known to be involved in cancer metastasis and various inflammatory responses.

[0174] As shown in Figure 9, neutrophil collagenase was confirmed to be specifically expressed in all MPNST tissues obtained from five patients. In particular, since neutrophil collagenase is secreted into the extracellular matrix, it is expected to be detectable in blood and urine samples, and is therefore expected to have a high utilization rate as an MPNST diagnostic marker.

[0175] Example 5. Discovery of a new therapeutic target for malignant peripheral nerve schwannoma (MPNST) 5.1. Hierarchical cluster analysis Proteomes that showed specifically increased expression in MPNST were divided into two clusters through hierarchical cluster analysis. More specifically, as shown in Figure 10a, of the 3,624 proteins identified in total in the two groups, 3,455 proteins were extracted from more than 80% of the samples in each group. Student's T test was performed on the two groups to select 292 proteins that satisfied Benjamini-Hochberg FDR < 0.01. Z scores were calculated for the quantitative information of these proteins, and K-means clustering analysis was performed to confirm the formation of two clusters. For the proteins in each cluster, g:Profiler (https: / / biit.cs.ut.ee / gprofiler / ) was used to investigate biological process, cellular compartment, and molecular function.

[0176] Referring to Figure 10b, Cluster 1 consists of proteins whose expression was increased in MPNST, and proteins that bind to the extracellular matrix and function, such as functional actin and musculoskeletal proteins, were discovered.

[0177] Referring to Figure 10c, Cluster 2 consists of proteins whose expression was reduced in MPNST, and which are mainly proteins secreted into the extracellular matrix or present in exosomes.

[0178] 5.2. Identification of target signaling pathways using GO analysis and IPA (ingenuity pathway analysis) Gene ontology analysis and QIAGEN ingenuity pathway analysis were performed on 144 MPNST-specific proteins to identify pathways involving proteins whose expression is specifically altered by MPNST. The accession numbers of the 144 MPNST-specific proteins were entered into ShynyGO and the IPA server, and the biological processes, cellular compartments, and molecular functions associated with these 144 MPNST-specific proteins were investigated.

[0179] As a result, as shown in Figures 11 and 12, we confirmed that the ILK signaling pathway, the actin-cytoskeleton signaling pathway, and the RhoGDI signaling pathway are specifically enhanced in MPNST. Furthermore, by inferring the higher-level signaling pathways of these signaling systems, we predicted that the enhancement of TGF-β is the underlying factor.

[0180] 5.3 Discovery of Kinases as Novel Therapeutic Targets The kinases that were specifically expressed or whose activity was increased in MPNST in relation to the signaling pathway discovered in Example 5.2 may be utilized as therapeutic targets. Therefore, the inventors discovered kinases that were increased in MPNST.

[0181] As a result, as shown in Figures 13 to 21, it was confirmed that kinases such as CDK4, MAP2K1, PDK3, STK10, ALPK3, PHKG1, CDK5, PNKP, and PACSIN3 were increased in MPNST. Therefore, inhibitors of these kinases, such as antagonists, may be used to treat MPNST.

[0182] In particular, fostamatinib exhibits multiple antagonistic effects on CDK4, MAP2k1, STK1, and PHKG1, and is therefore expected to be useful as a treatment for MPNST.

[0183] Example 6. Verification of diagnostic markers for malignant peripheral nerve schwannoma (MPNST) In order to verify the expression of MMP8 protein, one of the MPNST diagnostic markers discovered in Example 4, IHC staining was performed on the tissues of MPNST patients.

[0184] As a result, as shown in Table 4 and Figure 22 below, it was confirmed that the MMP8 protein (neutrophil collagenase) was overexpressed in MPNST tissue compared to NF tissue.

[0185] [Table 4]

[0186] Furthermore, differences in MMP8 protein expression in blood, which is easily obtained from patient samples, were confirmed using ELISA analysis. Specifically, ELISA analysis was performed on plasma from the normal group (WT00), NF1 patients (SEL00), MPNST patients who had their tumor site completely removed after surgery (post-OP) (clear), patients whose tumor was not completely removed due to the nature of tumor removal and who may still have tumors (intermediate), and patients whose tumors were not removed at all or who had metastases (involved).

[0187] As a result, as shown in Table 5 and Figure 23 below, it was confirmed that the expression of the MMP8 protein increased in the plasma of patients whose tumors were either not completely removed (intermediate) or had metastasized or not removed at all (involved). Based on these results, which confirmed the difference in MMP8 protein expression depending on whether or not tumors were removed, it was confirmed that MMP8 can be used as a diagnostic marker for MPNST.

[0188] [Table 5]

[0189] Example 7. Verification of therapeutic targets for malignant peripheral nerve schwannoma (MPNST) To demonstrate the abnormal activity of the ILK signaling pathway, the actin-cytoskeleton signaling pathway, and various kinases—MPNST therapeutic targets discovered in Example 5—IHC staining of ILK, Cofilin, MEK, p-MEK, and PACSIN3 proteins was performed in MPNST patient tissue.

[0190] As a result, as shown in Table 6 and Figure 24 below, it was confirmed that the expression of ILK, Cofilin, MEK, p-MEK, and PACSIN3 proteome was increased in MPNST tissue compared to NF tissue.

[0191] [Table 6]

[0192] The above-mentioned description of the present invention is illustrative, and a person with ordinary skill in the art to which the invention pertains will understand that it can be readily modified into other specific forms without altering the technical idea or essential features of the invention. Therefore, the above-mentioned embodiments should be understood to be illustrative in all respects and not limiting. [Industrial applicability]

[0193] The present invention relates to a composition for the differential diagnosis of malignant peripheral nerve sheath tumor (MPNST), and is expected to be useful in rapidly predicting and diagnosing the presence or absence of malignant tumors in patients with neurofibromatosis type 1, using a composition capable of detecting the protein marker of the present invention, thus having industrial applicability.

Claims

1. A composition for the differential diagnosis of malignant peripheral nerve sheath tumor (MPNST), comprising endosialin or a preparation for measuring its mRNA expression level as an active ingredient, Here, in the differential diagnosis of MPNST, a differential diagnosis composition is used to distinguish between neurofibromatosis type I (NF-1) and MPNST.

2. The composition for differentiating and determining malignant peripheral nerve sheaths according to claim 1, characterized in that the preparation for measuring the expression level of the endosialin protein is an antibody or aptamer specific to the protein.

3. The composition for differentiating malignant peripheral nerve sheath tumors according to claim 1, characterized in that the preparation for measuring the expression level of the mRNA is a primer set or probe that specifically binds to the mRNA.

4. A kit for the differential diagnosis of malignant peripheral nerve schwannoma, comprising the composition described in any one of claims 1 to 3, Here, in the differential diagnosis of MPNST, there is a differential diagnosis kit that distinguishes between neurofibromatosis type 1 (NF-1) and MPNST.

5. The kit for the differential diagnosis of malignant peripheral schwannoma according to claim 4, characterized in that the kit includes instructions indicating that if the expression level of endosialin or its mRNA is detected, or if the expression level is higher than that of a benign neurofibroma sample, it should be determined to be a malignant peripheral schwannoma.

6. A method for providing information necessary for the differential diagnosis of malignant peripheral nerve schwannoma, comprising the step of measuring the expression level of endosialin or its mRNA in a biological sample isolated from a subject, Here, we will discuss the method for differentiating MPNST from neurofibromatosis type 1 (NF-1).

7. The method for providing information necessary for the differential diagnosis of malignant peripheral nerve schwannoma according to claim 6, further comprising the step of confirming whether the expression of endosialin or its mRNA is detected, or whether the expression level thereof is higher than the expression level in a biological sample isolated from a patient with neurofibromatosis type 1.

8. The method for providing information necessary for the differential diagnosis of malignant peripheral nerve schwannoma according to claim 6, characterized in that the subject is a patient suspected of having malignant peripheral nerve schwannoma or a patient determined to have neurofibromatosis type 1.

9. The method for providing information necessary for the differential diagnosis of malignant peripheral nerve schwannoma according to claim 6, characterized in that the biological sample is tissue.

10. The method for providing information necessary for the differential diagnosis of malignant peripheral nerve schwannomas according to claim 6, characterized in that the endosialin protein expression level is measured by at least one method selected from the group consisting of Western blotting, ELISA, radioimmunoanalysis, radioimmunodiffusion, Ochterlony immunodiffusion, Rocket immunoelectrophoresis, immunostaining, immunoprecipitation analysis, complement fixation analysis, mass spectrometry, FACS, and protein chips.

11. The method for providing information necessary for the differential diagnosis of malignant peripheral nerve schwannoma according to claim 6, characterized in that the mRNA expression level is measured by at least one method selected from the group consisting of PCR, RNase protective analysis, northern blotting, southern blotting, in situ cross-pollination, and DNA chip.

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