KR102089371B1 - Markers for diagnosis angioimmunoblastic T cell lymphoma and uses thereof - Google Patents

Abstract

The present invention relates to a marker for diagnosing vascular immunocytoblastic T cell lymphoma and uses thereof, and more specifically, vascular immunocytoblast including CTLA4 (cytotoxic T-lymphocyte associated protein 4) and CD28 gene or a protein encoded by the gene. A composition for diagnosing T cell lymphoma, a composition for diagnosing vascular immunocytoblastic T cell lymphoma comprising an agent for measuring the gene or protein level, and a method for providing information for diagnosing vascular immunocytoblastic T cell lymphoma.
Vascular immunocytoblastic T cell lymphoma diagnosis technology according to the present invention accurately diagnoses vascular immunocytoblastic T-cell lymphoma while minimizing the possibility of misdiagnosis of other subtypes by measuring the expression level or copy number of CTLA4 and CD28 genes. As it can be done, it is expected to be usefully used to enhance the therapeutic effect of vascular immunoblastic T-cell lymphoma.

Description

Markers for diagnosis angioimmunoblastic T cell lymphoma and uses thereof

The present invention relates to a marker for diagnosing vascular immunocytoblastic T cell lymphoma and uses thereof, and more specifically, a cytotoxic T-lymphocyte associated protein 4 (CTLA4) and a CD28 gene or a protein encoded by the gene, an vascular immunoblast cell. A marker composition for diagnosing sex T cell lymphoma, an agent for measuring the gene or protein level, a composition for diagnosing vascular immunocytoblastic T cell lymphoma, and a method for providing information for diagnosing vascular immunocytoblastic T cell lymphoma.

Peripheral T-cell lymphoma is a malignant tumor of T-cells, of which angioimmunoblstic T-cell lymphoma (AITL) is a major T-caused T cell invading lymph nodes. Among the lymphomas of the cells, they are derived from CD4 antigen-positive lymphocytes that support immune function in peripheral blood (peripheral CD4 + helper T-cell), but are generally a rare but relatively common lymphoma among peripheral T-cell lymphomas.

Anemia, eosinophilia, showing clinical features such as systemic lymph node swelling, hepatomegaly, and fever and skin lesions, mainly showing partial loss of normal lymph node structure and polymorphic cell infiltration and vascular proliferation. Coombs' test shows characteristics accompanied by immunological abnormalities such as positive and hypergamma globulinemia. Vascular immunocytoblastic T-cell lymphoma has a poor prognosis compared to other subtypes of non-Hodgkin's lymphoma with a 5-year survival rate of 30-35% and a median survival time of less than 36 months, although there are some differences among reporters. Although there are histological findings, it is pointed out that the diagnosis is delayed or misdiagnosed as a significant morphological overlap with other subtypes of malignant lymphoma, and there is no characteristic phenotypic or molecular marker of the disease.

Recently, studies on the characteristic ecological markers of malignant T-cells in vascular immunocytoblastic T-cell lymphomas have been actively conducted to understand the ecology of this disease and to discriminate it from other diseases (Attygalle A, Al-Jehani). R, Diss TC, et al. Neoplastic T cells in angioimmunoblastic T-cell lymphoma express CD10.Blood 2002; 99: 627-33), in Korea due to low disease frequency of vascular immunocytoblastic T-cell lymphoma. There are insufficient reports on the clinical characteristics, treatment outcomes, and prognostic factors unique to Korea.

Therefore, the method of accurately diagnosing vascular immunocytoblastic T-cell lymphoma while minimizing the possibility of misdiagnosis of other subtypes is very important, so research and technology development are required.

The present inventors have conducted extensive research to discover biomarkers capable of diagnosing angioimmunoblastic T cell lymphoma (AITL), and the number of copies of each gene of CTLA4 and CD28 in an angiogenic immunocytoblastic T cell lymphoma patient ( The present invention was completed by confirming the increase in copy number and overexpression of each gene.

Accordingly, an object of the present invention is to provide a marker composition for diagnosing vascular immunocytoblastic T cell lymphoma, comprising CTLA4 (cytotoxic T-lymphocyte associated protein 4) and CD28 gene, or a protein encoded by the gene.

In addition, another object of the present invention is to provide a composition for diagnosing vascular immunocytoblastic T cell lymphoma, comprising an agent for measuring the expression level of mRNA of CTLA4 and CD28 genes or a protein encoded by the gene.

In addition, the present invention relates to a biological sample derived from a human subject, i) measuring the mRNA level of the CTLA4 and CD28 genes, or the protein level encoded by the gene, or ii) measuring the copy number of the CTLA4 and CD28 genes. Another object is to provide an information providing method for diagnosing vascular immunocytoblastic T cell lymphoma.

In addition, the present invention

(1) processing the candidate substance in cells in vitro;

(2) in the cell i) measuring the expression level of mRNA or protein of CTLA4 and CD28 genes or ii) measuring the copy number of CTLA4 and CD28 genes; And

(3) selecting a substance that reduces the expression level of the mRNA or protein of the CTLA4 and CD28 genes, or the copy number of the CTLA4 and CD28 genes, as a therapeutic substance, compared to a non-treatment group of candidate substances Another object of the present invention is to provide a method for screening for therapeutic agents for vascular immunocytoblastic T cell lymphoma.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems that are not mentioned will be clearly understood by those skilled in the art from the following description.

To achieve the above object, the present invention provides a marker composition for diagnosing vascular immunocytoblastic T cell lymphoma, comprising a cytotoxic T-lymphocyte associated protein 4 (CTLA4) and a CD28 gene, or a protein encoded by the gene.

In addition, the present invention provides a composition for diagnosing vascular immunocytoblastic T cell lymphoma, comprising an agent that measures the expression level of the mRNA of the cytotoxic T-lymphocyte associated protein 4 (CTLA4) and the CD28 gene or the protein encoded by the gene. .

In one embodiment of the present invention, the agent for measuring the level of mRNA of the gene may be a sense and antisense primer, or a probe that complementarily binds to the mRNA of the gene.

In another embodiment of the present invention, an agent that measures the level of the protein may be an antibody specifically defective in the protein.

In addition, the present invention provides a kit for diagnosing vascular immunocytoblastic T cell lymphoma comprising the composition.

In addition, the present invention relates to a biological sample derived from a human subject, i) measuring the mRNA level of the CTLA4 and CD28 genes, or the protein level encoded by the gene, or ii) measuring the copy number of the CTLA4 and CD28 genes. It provides a method for providing information for diagnosing vascular immunocytoblastic T cell lymphoma.

In one embodiment of the present invention, the mRNA level or the number of copies is measured through a method of polymerase chain reaction (PCR), reverse transcriptase chain reaction (RT-PCR) or real-time polymerase chain reaction (Real-time PCR). Can be.

In another embodiment of the present invention, the protein expression level is western blotting, radioimmunoassay (RIA), radioimmunodiffusion, enzymatic immunoassay (ELISA), immunoprecipitation (immunoprecipitation). , Flow cytometry, or immunofluorescence staining (immunofluorescnce).

In addition, the present invention

(1) processing the candidate substance in cells in vitro;

(2) in the cell i) measuring the expression level of mRNA or protein of CTLA4 and CD28 genes or ii) measuring the copy number of CTLA4 and CD28 genes; And

(3) selecting a substance that reduces the expression level of the mRNA or protein of the CTLA4 and CD28 genes, or the copy number of the CTLA4 and CD28 genes, as a therapeutic substance, compared to a non-treatment group of candidate substances The present invention provides a method for screening for therapeutic agents for vascular immunocytoblastic T cell lymphoma.

In one embodiment of the invention, the candidate material may be selected from the group consisting of nucleic acids, compounds, microbial cultures or extracts, natural product extracts, peptides, substrate analogs, aptamers, and antibodies.

In another embodiment of the present invention, the nucleic acid is a group consisting of siRNA, shRNA, microRNA, antisense RNA, aptamer, locked nucleic acid (LNA), peptide nucleic acid (PNA), and morpholino. Can be selected from.

Vascular immunocytoblastic T cell lymphoma diagnosis technology according to the present invention accurately diagnoses vascular immunocytoblastic T-cell lymphoma while minimizing the possibility of misdiagnosis of other subtypes by measuring the expression level or copy number of CTLA4 and CD28 genes. As it can be done, it is expected to be usefully used to increase the therapeutic effect of vascular immunoblastic T-cell lymphoma.

1A and 1B are results of confirming a change in the copy number of CTLA4 and CD28 in AITL patients through quantitative-PCR.
2 is a result of confirming the change in the expression level of CTLA4 and CD28 in AITL patients through quantitative-PCR.

The present invention provides a marker composition for diagnosing vascular immunocytoblastic T cell lymphoma, comprising CTLA4 (cytotoxic T-lymphocyte associated protein 4) and CD28 gene, or a protein encoded by the gene.

In addition, the present invention includes a composition for measuring the expression level of mRNA of the CTLA4 (cytotoxic T-lymphocyte associated protein 4) and CD28 gene or a protein encoded by the gene, and a composition for diagnosing vascular immunocytoblastic T cell lymphoma and the same Provides a kit for diagnosing angiogenic immunocytoblastic T cell lymphoma.

The present inventors have found that the copy number and expression level of each gene of CTLA4 and CD28 are closely related to vascular immunocytoblastic T cell lymphoma.

In an embodiment of the present invention, as a result of measuring the copy number of the CTLA4 and CD28 genes in a sample of an AITL patient, the CTLA4 and CD28 genes in the case of a fusion-positive AITL patient expressing the CD-28-CTLA4 fusion gene It was confirmed that the number of copies (copy number) was significantly increased (Example 1).

In another embodiment of the present invention, as a result of confirming the mRNA expression levels of CTLA4 and CD28 in a sample of an AITL patient, mRNA expression of CD28 and CTLA4 was significantly increased in a fusion-positive AITL patient expressing a CD-28-CTLA4 fusion gene. It was confirmed to increase (Example 2).

Through the above results, it can be seen that the CTLA4 and CD28 genes or proteins encoded by the genes can be used as markers for diagnosis of vascular immunocytoblastic T cell lymphoma.

In the present invention, the agent for measuring the mRNA level of the CTLA4 and CD28 genes may be sense and antisense primers or probes that complementarily bind to mRNA, but is not limited thereto.

The term "primer" used in the present invention is a short gene sequence serving as a starting point for DNA synthesis, and refers to an oligonucleotide synthesized for the purpose of diagnosis, DNA sequencing, and the like. The primers can generally be synthesized to a length of 15 to 30 base pairs, but may vary depending on the purpose of use, and can be modified by methylation, capping, or the like by a known method.

As used in the present invention, the term "probe" refers to a nucleic acid capable of specifically binding to mRNA of several bases to hundreds of bases produced through enzymatic chemical separation or synthesis. The presence or absence of mRNA can be confirmed by labeling radioactive isotopes or enzymes, and can be designed and modified in a known manner.

The agent for measuring the protein level may be an antibody that specifically binds to a protein encoded by a gene, but is not limited thereto.

As used herein, the term “antibody” includes immunoglobulin molecules that are immunologically reactive with a specific antigen, and includes both monoclonal and polyclonal antibodies. In addition, the antibody includes forms produced by genetic engineering such as chimeric antibodies (eg, humanized murine antibodies) and heterologous antibodies (eg, bispecific antibodies).

The kit for diagnosing vascular immunocytoblastic T cell lymphoma of the present invention may be composed of one or more other component compositions, solutions, or devices suitable for analytical methods.

For example, in order to perform PCR, the kit of the present invention contains genomic DNA derived from a sample to be analyzed, a primer set specific for the marker gene of the present invention, an appropriate amount of DNA polymerase, dNTP mixture, PCR buffer solution and water. It may be a kit including. The PCR buffer solution may contain KCl, Tris-HCl and MgCl2. In addition, components necessary for performing electrophoresis capable of confirming whether or not PCR products are amplified may be additionally included in the kit of the present invention.

In addition, the kit of the present invention may be a kit including essential elements necessary for performing RT-PCR. RT-PCR kits include enzymes such as test tubes or other suitable containers, reaction buffers, deoxynucleotides (dNTPs), Taq-polymerases and reverse transcriptases, DNase, RNase inhibitors, DEPCs, in addition to each primer pair specific for a marker gene. -It may include DEPC-water, sterile water, and the like. In addition, it may include a primer pair specific to the gene used as a quantitative control.

In addition, the kit of the present invention may be a kit containing essential elements necessary to perform a DNA chip. The DNA chip kit includes a substrate to which a cDNA corresponding to a gene or a fragment thereof is attached as a probe, and the substrate may include a cDNA corresponding to a quantitative structural gene or a fragment thereof. In addition, the kit of the present invention may be in the form of a microarray having a substrate on which the marker gene of the present invention is immobilized.

In another aspect of the present invention, the present invention relates to a biological sample derived from a human subject, i) measuring the mRNA level of the CTLA4 and CD28 genes, or the protein level encoded by the gene, or ii) the number of copies of the CTLA4 and CD28 genes ( It provides a method for providing information for the diagnosis of vascular immunocytoblastic T cell lymphoma, including the step of measuring a copy number).

The expression level and the number of copies of the mRNA can be measured through methods such as polymerase chain reaction (PCR), reverse transcriptase chain reaction (RT-PCR), and real-time polymerase chain reaction in a conventional manner known in the art. , But is not limited to this.

The protein expression level is Western blotting, radioimmunoassay (RIA), radioimmunodiffusion, enzymatic immunoassay (ELISA), immunoprecipitation, by conventional methods known in the art. , Flow cytometry, immunofluorescence, and the like, but it is not limited thereto.

As another aspect of the present invention, the present invention provides a method for screening for treating an angiogenic immunocytoblastic T cell lymphoma comprising the following steps.

(1) processing the candidate substance in cells in vitro;

(2) in the cells i) measuring the expression level of mRNA or protein of CTLA4 and CD28 genes, or ii) measuring the copy number of CTLA4 and CD28 genes; And

(3) Comprising the step of selecting a substance that reduces the expression level of the mRNA or protein of the CTLA4 and CD28 genes, or the copy number of the CTLA4 and CD28 genes as a therapeutic substance, compared to the untreated group of candidate substances , Vascular immunoblastic T cell lymphoma therapeutic screening method.

The candidate material may be selected from the group consisting of compounds, microbial cultures or extracts, natural product extracts, nucleic acids, and peptides, and the nucleic acids are preferably siRNA, shRNA, microRNA, antisense RNA, aptamer, LNA ( It may be selected from the group consisting of locked nucleic acid (PNA), peptide nucleic acid (PNA), and morpholino, but is not limited thereto.

Hereinafter, preferred embodiments are provided to help understanding of the present invention. However, the following examples are only provided to more easily understand the present invention, and the contents of the present invention are not limited by the following examples.

[Example]

Example 1. Patient sample preparation

All AITL patient samples were obtained under the approval of the Helsinki Declaration and Samsung Medical Center System Review Committee, and the clinical information of specific AITL patients is shown in Table 1 below.

case no. Diagnosis Age Gender Tissue of origin CD-28-CTLA4 fusion One AITL 66 M Lymph node N 2 AITL 64 M Lymph node N 3 AITL 28 F Lymph node P 4 AITL 59 M Lymph node P 5 AITL 76 F Lymph node P 6 AITL 72 M Lymph node P 7 AITL 67 M Lymph node P 8 AITL 59 M Lymph node P 9 AITL 52 M Lymph node P 10 AITL 42 M Lymph node P 11 AITL 60 F Lymph node N 12 AITL 53 F Lymph node P 13 AITL 67 M Lymph node N 14 AITL 57 F Lymph node P 15 AITL 58 M Lymph node N 16 AITL 62 M Lymph node P 17 AITL 51 M Lymph node P 18 AITL 60 F Lymph node P 19 AITL 57 F Lymph node P 20 AITL 47 M Lymph node P 21 AITL 50 F Lymph node N 22 AITL 58 M Lymph node N 23 AITL 67 M Lymph node N 24 AITL 75 M Lymph node P 25 AITL 64 M Lymph node N 26 AITL 74 M Lymph node P 27 AITL 72 M Lymph node N 28 AITL 75 M Lymph node N 29 AITL 62 M Lymph node P 30 AITL 38 M Lymph node N 31 AITL 54 M Lymph node P 32 AITL 57 M Lymph node P 33 AITL 65 M Lymph node P 34 AITL 41 F Lymph node N 35 AITL 77 M Lymph node N 36 AITL 78 M Lymph node N 37 AITL 79 M Lymph node N 38 AITL 57 M Lymph node P 39 AITL 73 F Lymph node N 40 AITL 70 M Lymph node P 41 AITL 63 M Lymph node P 42 AITL 60 F Lymph node P 43 AITL 77 M Lymph node N 44 AITL 56 M Lymph node N 45 AITL 48 M Lymph node N

Example  2. AITL  From the patient CTLA4  And CD28 Number of copies (copy number) Confirm change

Changes in the copy number of CTLA4 and CD28 genes in samples of AITL patients prepared in Example 1 were confirmed by quantitative-PCR. At this time, the specific information of the primer used is shown in Table 2 below.

division Sequence information (5'- 3 ') Sequence number CD28_gQ1
F 5'-AGGCATTGATGAGGATACGC-3 ' One R 5'-TTCTATCCCTTGCCATGACC-3 ' 2 CD28_gQ2
F 5'-AGGGATGGGTTACAGCACAG-3 ' 3 R 5'-GAGTTCGAGGAAGCCAGTTG-3 ' 4 CD28_gQ3
F 5'-CGGTGAGCAAGCAGAATACA-3 ' 5 R 5'-GGAAGAGCAACCAACTCCAG-3 ' 6 CD28_gQ4
F 5'-GGCCCACATTCCAACTTACC-3 ' 7 R 5'-GGGAAGAGGCTCCCAGAATC-3 ' 8 CD28_gQ5
F 5'-TCCAATCAGACCAGGTAGGAGC-3 ' 9 R 5'-CCACAACCCACTTTGGATCTCC-3 ' 10 CD28_gQ6
F 5'-CACAGGCATGTTCCTACCTCAGG-3 ' 11 R 5'-GGACCTGAAGGGTGACGAGG-3 ' 12 CTLA4_gQ1
F 5'-CTCACTATCCAAGGACTGAGGGC-3 ' 13 R 5'-CTGGGTTCCGTTGCCTATGC-3 ' 14 CTLA4_gQ2
F 5'-TGCAATTTAGGGGTGGACCT-3 ' 15 R 5'-AGAATCTGGGCACGGTTCTGGAT-3 ' 16 CTLA4_gQ3
F 5'-CCATCACCTGGAAGTCACCT-3 ' 17 R 5'-CCCAGTCAAGCAAACTGGAT-3 ' 18 CTLA4_gQ4
F 5'-CAGGGAAGTTTTGTGGAGGA-3 ' 19 R 5'-CACAATTCCACGCAATCAAG-3 ' 20 β-actin_gQ
F 5'-TGAGCCTCATCTCCCACGTA-3 ' 21 R 5'-TCTCAGCCAGCACCATGACT-3 ' 22

As a result, as shown in Figure 1a and 1b, it was confirmed that the copy number (copy number) of the CTLA4 and CD28 genes in the samples of AITL patients increased. In particular, in the case of a fusion-positive AITL patient expressing the CD-28-CTLA4 fusion gene, it was confirmed that the copy number of the CTLA4 and CD28 genes increased significantly.

Example 3. Confirmation of CTLA4 and CD28 expression level changes in AITL patients

In the samples of AITL patients prepared in Example 1, mRNA expression levels of CTLA4 and CD28 genes were confirmed through quantitative-PCR. At this time, the specific information of the primer used is shown in Table 3 below.

division SEQ ID NO (5'-3 ') Sequence number CD28_cQ
F 5'-ACAATGCGGTCAACCTTAGC-3 ' 23 R 5'-ACCTGAAGCTGCTGGGAGTA-3 ' 24 CTLA4_cQ
F 5'-GTGCCCAGATTCTGACTTCC-3 ' 25 R 5'-CTGGCTCTGTTGGGGGCATTTTC-3 ' 26 Fusion_cQ
F 5'-CAGCAGTTAGTTCGGGGTTG-3 ' 27 R 5'-GCGGGGAGTCATGTTCATGT-3 ' 28 β-actin_cQ
F 5'-CCAACCGCGAGAAGATGACC-3 ' 29 R 5'-GGTCCAGACGCAGGATGGC-3 ' 30

As a result, as shown in Figure 2, it was confirmed that the mRNA expression of CTLA4 and CD28 is increased in samples of AITL patients. Particularly, in the case of a fusion-positive AITL patient expressing the CD-28-CTLA4 fusion gene, it was confirmed that the mRNA expression of CD28 and CTLA4 increased by about 3 and 7 times, respectively, in the fusion-positive patient.

The above description of the present invention is for illustration only, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

<110> SAMSUNG LIFE PUBLIC WELFARE FOUNDATION <120> Markers for diagnosis angioimmunoblastic T cell lymphoma and uses          thereof <130> PD17-009 <160> 30 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ1 F <400> 1 aggcattgat gaggatacgc 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ1 R <400> 2 ttctatccct tgccatgacc 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ2 F <400> 3 agggatgggt tacagcacag 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ2 R <400> 4 gagttcgagg aagccagttg 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ3 F <400> 5 cggtgagcaa gcagaataca 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ3 R <400> 6 ggaagagcaa ccaactccag 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ4 F <400> 7 ggcccacatt ccaacttacc 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ4 R <400> 8 gggaagaggc tcccagaatc 20 <210> 9 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ5 F <400> 9 tccaatcaga ccaggtagga gc 22 <210> 10 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ5 R <400> 10 ccacaaccca ctttggatct cc 22 <210> 11 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ6 F <400> 11 cacaggcatg ttcctacctc agg 23 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CD28_gQ6 R <400> 12 ggacctgaag ggtgacgagg 20 <210> 13 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> CTLA4_gQ1 F <400> 13 ctcactatcc aaggactgag ggc 23 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CTLA4_gQ1 R <400> 14 ctgggttccg ttgcctatgc 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CTLA4_gQ2 F <400> 15 tgcaatttag gggtggacct 20 <210> 16 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> CTLA4_gQ2 R <400> 16 agaatctggg cacggttctg gat 23 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CTLA4_gQ3 F <400> 17 ccatcacctg gaagtcacct 20 <210> 18 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CTLA4_gQ3 R <400> 18 cccagtcaag caaactggat 20 <210> 19 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CTLA4_gQ4 F <400> 19 cagggaagtt ttgtggagga 20 <210> 20 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CTLA4_gQ4 R <400> 20 cacaattcca cgcaatcaag 20 <210> 21 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> beta-actin_gQ F <400> 21 tgagcctcat ctcccacgta 20 <210> 22 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> beta-actin_gQ R <400> 22 tctcagccag caccatgact 20 <210> 23 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CD28_cQ F <400> 23 acaatgcggt caaccttagc 20 <210> 24 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CD28_cQ R <400> 24 acctgaagct gctgggagta 20 <210> 25 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CTLA4_cQ F <400> 25 gtgcccagat tctgacttcc 20 <210> 26 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> CTLA4_cQ R <400> 26 ctggctctgt tgggggcatt ttc 23 <210> 27 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Fusion_cQ F <400> 27 cagcagttag ttcggggttg 20 <210> 28 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Fusion_cQ R <400> 28 gcggggagtc atgttcatgt 20 <210> 29 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> beta-actin_cQ F <400> 29 ccaaccgcga gaagatgacc 20 <210> 30 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> beta-actin_cQ R <400> 30 ggtccagacg caggatggc 19

Claims (11)

delete i) an agent that measures the copy number or mRNA of the CTLA4 gene or the expression level of the protein encoded by the gene; And
ii) A composition for diagnosing vascular immunocytoblastic T cell lymphoma having a CTLA4 and CD28 fusion gene, comprising an agent that measures the copy number or mRNA of a CD28 gene or the expression level of a protein encoded by the gene.
According to claim 2,
The agent for measuring the level of mRNA of the gene is characterized in that the sense and antisense primers, or probes that complementarily bind to the mRNA of the gene, diagnostic composition.
According to claim 2,
The agent for measuring the level of the protein is characterized in that the antibody specifically binding to the protein, diagnostic composition.
A kit for diagnosing vascular immunocytoblastic T cell lymphoma having CTLA4 and CD28 fusion genes, comprising the composition of claim 2.
For biological samples derived from human subjects,
i) the mRNA of CTLA4 and CD28 genes, or the protein level encoded by the gene is measured or
ii) A method of providing information for diagnosing vascular immunocytoblastic T cell lymphoma with CTLA4 and CD28 fusion genes, comprising the step of measuring the copy number of CTLA4 and CD28 genes.
The method of claim 6,
The mRNA level or the number of copies is measured through the method of polymerase chain reaction (PCR), reverse transcriptase chain reaction (RT-PCR), or real-time polymerase chain reaction (Real-time PCR), information How to provide.
The method of claim 6,
The protein expression level is western blotting, radioimmunoassay (RIA), radioimmunodiffusion, enzyme immunoassay (ELISA), immunoprecipitation, flow cytometry, Or characterized by being measured through immunofluorescence staining (immunofluorescnce), information providing method.
(1) processing the candidate substance on the cell in vitro;
(2) in the cells i) measuring the expression level of mRNA or protein of CTLA4 and CD28 genes or ii) measuring the copy number of CTLA4 and CD28 genes; And
(3) selecting a substance that reduces the expression level of the mRNA or protein of the CTLA4 and CD28 genes, or the copy number of the CTLA4 and CD28 genes, as a therapeutic substance, compared to a non-treatment group of candidate substances A method for screening for therapeutic agents for vascular immunocytoblastic T cell lymphoma having CTLA4 and CD28 fusion genes.
The method of claim 9,
The candidate material is selected from the group consisting of nucleic acid, compound, microbial culture or extract, natural product extract, peptide, substrate analog, aptamer, and antibody, screening method.
The method of claim 10,
The nucleic acid is selected from the group consisting of siRNA, shRNA, microRNA, antisense RNA, aptamer, locked nucleic acid (LNA), peptide nucleic acid (PNA), and morpholino, Screening method.

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