JP6667790B2 - Test method for T cell lymphoma - Google Patents

Description

  The present invention relates to a method for testing T cell lymphoma, which is a novel molecular diagnostic method, and a method for screening a therapeutic agent for T cell lymphoma based on the molecular pathology of T cell lymphoma.

  T-cell lymphoma is part of non-Hodgkin's lymphoma and is classified as Peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS) and adult T-cell leukemia / lymphoma (adult T-cell leukemia / It is a group of diseases including various types such as lymphoma (ATL). Among them, ATL is a lymphoma caused by HTLV-1 virus infection, and is known to be an extremely poor prognosis. The incubation period is long from the infection of HTLV-1 virus to the onset of ATL, and the protein Tax encoded by the HTLV-1 virus is inactivated at the onset of ATL. These findings suggest that ATL development is related to somatic mutations acquired after HTLV-1 virus infection. However, the overall picture of the genetic basis of T-cell lymphoma, including ATL, is poorly understood.

  Currently, T cell lymphoma is diagnosed by pathological diagnosis by lymph node biopsy and flow cytometry when symptoms and signs such as swelling of lymph nodes are observed and T cell lymphoma is suspected. In addition, ATL is diagnosed by confirming HLTV-1 integration by HTLV-1 antibody test or Southern blot. T-cell lymphoma is a disease that is resistant to conventional chemotherapy, and radical treatment is limited to hematopoietic stem cell transplantation. However, hematopoietic stem cell transplantation is a treatment with a large number of treatment-related deaths and is not indicated for the elderly or patients with complications. The treatment of ATL is selected from chemotherapy with anticancer drugs, hematopoietic stem cell transplantation, and the like. Recently, a new anti-CCR4 antibody against CC chemokine receptor 4 (CCR4) has been approved for CCR4-positive ATL. However, at present, treatment methods for T cell lymphoma, mainly ATL, are limited, and there is an eager desire to understand the molecular pathology of T cell lymphoma and to develop pathological-specific treatments.

  For ATL, it has been reported that diagnosis and prognosis can be performed by measuring the methylation status of CpG islands in the promoter region such as the SHP1 gene (Patent Documents 1 and 2). In addition, as a comprehensive genetic analysis of T-cell lymphoma, as a result of comprehensive genetic analysis of PTCL-NOS and angioimmunoblastic T-cell lymphoma (AITL), the RHOA gene was analyzed. It has been reported by the present inventors that mutations are frequently observed (Non-patent document 1: Nat Genet. 2014; 46 (2): 166-70, Non-patent document 2: Nat Genet. 2014; 46 ( 2): 171-5, Non-Patent Document 3: Nat Genet. 2014; 46 (4): 371-5). As a genetic analysis of ATL, TP53 gene (Non-patent document 4: Blood. 1992; 79 (2): 477-80) and FAS gene (Non-patent document 5: Blood. 1998; 91 (10): 3935-42.), NOTCH1 gene (Non-patent document 6: Proc Natl Acad Sci US A. 2010; 107 (38): 16619-24.), JAK3 gene (Non-patent document 7: Blood. 2011). Oct 6; 118 (14): 3911-21) and CDKN2A gene (Non-patent document 8: J Clin Oncol. 1997; 15 (5): 1778-85) or ZEB1 by copy number analysis of the gene. Gene (Non-patent document 9: Blood. 2008; 112 (2): 383-93), NDRG2 gene (Non-patent document 10: Nat Commun. 2014; 5: 3393), miR-31 (Non-patent document 11: Cancer Cell) 2012; 21 (1): 121-35), amplification of CARD11 gene and BCL11B gene (Non-Patent Document 12: Blood. 2006; 107 (11): 4500-7). I have.

  In the above report, genetic analysis was performed on a small number of cases of PTCL-NOS and AITL, and gene abnormalities depending on individual cases were detected, but the overall picture of genetic abnormalities in T cell lymphoma remains unknown It is. In addition, the functional significance of many of the identified genetic abnormalities remains to be elucidated, and has not led to the understanding of molecular pathologies or the development of pathological-specific treatments. Comprehensive genetic analysis using a large number of cases is desired to clarify the overall picture of genetic abnormalities in T-cell lymphoma, but this has not been achieved.

JP 2007-244377 A JP 2009-254357 A

Nat Genet. 2014; 46 (2): 166-70 Nat Genet. 2014; 46 (2): 171-5 Nat Genet. 2014; 46 (4): 371-5 Blood. 1992; 79 (2): 477-80 Blood. 1998; 91 (10): 3935-42 Proc Natl Acad Sci U S A. 2010; 107 (38): 16619-24 Blood. 2011 Oct 6; 118 (14): 3911-21 J Clin Oncol. 1997; 15 (5): 1778-85 Blood. 2008; 112 (2): 383-93 Nat Commun. 2014; 5: 3393 Cancer Cell. 2012; 21 (1): 121-35 Blood. 2006; 107 (11): 4500-7

  An object of the present invention is to elucidate the molecular pathology of T-cell lymphoma and to provide a new T-cell lymphoma test method and therapeutic target.

  The present inventors, in order to solve the above problems, using a T-cell lymphoma specimen for more than 350 cases, when comprehensively analyzing the genetic abnormality that occurs frequently, T-cell lymphoma specimen, Focusing on the fact that high-frequency gene mutations are detected in 35 kinds of genes including the PRKCB gene and CCR4 gene, in particular, by detecting gene mutations for at least one or more of the PRKCB gene and CCR4 gene It has been found that a test for T cell lymphoma can be performed, and the present invention has been completed.

That is, the present invention includes the following.
1. A test method for T cell lymphoma, comprising a step of detecting a mutation in at least one of PRKCB gene and CCR4 gene in a sample collected from a subject.
2. 2. The method for testing a T-cell lymphoma according to the above item 1, comprising a step of detecting a mutation in both the PRKCB gene and the CCR4 gene in a sample collected from a subject.
3. The T cell lymphoma according to the above 1 or 2, further comprising a step of detecting a mutation in at least one or more of the PLCG1 gene, CARD11 gene, CCR7 gene, and STAT3 gene for the specimen collected from the subject. Inspection method.
4. 4. The method for testing a T-cell lymphoma according to the above item 3, further comprising a step of detecting a mutation in all of the PLCG1, CARD11, CCR7, and STAT3 genes in a sample collected from the subject.
5. For samples collected from subjects, furthermore, TP53 gene, NOTCH1 gene, VAV1 gene, TBL1XR1 gene, GATA3 gene, IRF4 gene, FAS gene, POT1 gene, RHOA gene, IRF2BP2 gene, TET2 gene, HNRNPA2B1 gene, EP300 gene , CD58 gene, GPR183 gene, CSNK1A1 gene, CSNK2B gene, FYN gene, B2M gene, CBLB gene, CD28 gene, ZFP36L2 gene, CSNK2A1 gene, ATXN1 gene, SYNCRIP gene, S1PR1 gene, RELA gene, CDKN2A gene, and ICOS gene 5. The method for testing a T cell lymphoma according to any one of the above items 1 to 4, comprising a step of detecting a mutation of at least one or more genes.
6. To assist in the classification of T-cell lymphoma types, including the further step of detecting a mutation in at least one of the STAT3, TP53, IRF4, and CD58 genes in a sample collected from a subject. The method for testing a T-cell lymphoma according to any one of the above items 1 to 5.
7. A method for assisting in predicting the prognosis of T-cell lymphoma, comprising a step of further detecting a mutation in at least one gene of STAT3 gene, TP53 gene, IRF4 gene, and CD58 gene in a sample collected from a subject. The method for testing a T-cell lymphoma according to any one of the above items 1 to 5.
8. Mutation of the PRKCB gene is caused by mutation of at least one or more amino acids at amino acids 427, 470, 533, 546 and 630 in the amino acid sequence of SEQ ID NO: 1. 8. The method for testing a T-cell lymphoma according to any one of the above items 1 to 7, which is a gene mutation that results.
9. The preceding claim, wherein the mutation of the CCR4 gene is a gene mutation that causes at least one or more nonsense mutations or frameshift mutations in the C-terminal region, which is the amino acid sequence from position 309 to position 360 in the amino acid sequence of SEQ ID NO: 3. 9. The method for testing a T cell lymphoma according to any one of 1 to 8.
10. A gene mutation in which a mutation in the CCR4 gene results in a mutation in at least one or more of the amino acids 322, 323, 330, 331, and 336 of SEQ ID NO: 3. 10. The method for testing a T-cell lymphoma according to any one of the above items 1 to 9,
11. The method for testing a T cell lymphoma according to any one of the above items 3 to 10, wherein the mutation of each gene is selected from the following:
(I) For the PLCG1 gene, the amino acid at position 48, the amino acid at position 345, the amino acid at position 520, the amino acid at position 748, the amino acid at position 1016, the amino acid at position 1163, and the amino acid at position 1165 in the amino acid sequence of SEQ ID NO: 5 Gene mutation that results in mutation of at least one or more amino acids:
(Ii) mutation of at least one amino acid of amino acid 357, amino acid 361, amino acid 401, amino acid 615, and amino acid 626 in the amino acid sequence of SEQ ID NO: 9 with respect to CARD11 gene Gene mutations that result in:
(Iii) a gene mutation that causes a mutation in at least one of the 349th amino acid, the 351st amino acid, and the 355th amino acid in the amino acid sequence of SEQ ID NO: 7 with respect to the CCR7 gene.
12. The method for testing a T cell lymphoma according to any one of the above items 5 to 11, wherein the mutation of each gene is selected from the following:
(1) With respect to the VAV1 gene, at least one of the 174th amino acid, the 175th amino acid, the 404th amino acid, the 556th amino acid, the 798th amino acid, and the 822th amino acid in the amino acid sequence of SEQ ID NO: 19 Gene mutations that result in the above amino acid mutations:
(2) For the GPR183 gene, a gene mutation that causes at least one or more nonsense mutation or frameshift mutation in the amino acid sequence of SEQ ID NO: 21:
(3) For the TBL1XR1 gene, a gene mutation that causes at least one or more missense mutation, nonsense mutation or frameshift mutation in the amino acid sequence of SEQ ID NO: 15:
(4) For the GATA3 gene, a gene mutation that causes at least one or more nonsense mutation, frameshift mutation or splice site mutation in the amino acid sequence of SEQ ID NO: 11:
(5) a gene mutation that causes a mutation of at least one amino acid of the amino acid at position 59, the amino acid at position 70, and the amino acid at position 114 in the amino acid sequence of SEQ ID NO: 13 with respect to the IRF4 gene:
(6) For the IRF2BP2 gene, a gene mutation that causes at least one or more missense mutation, nonsense mutation or frameshift mutation in the amino acid sequence set forth in SEQ ID NO: 23:
(7) A gene mutation that causes at least one mutation of a splice site mutation of exon 10 and a mutation of the 322nd amino acid in the amino acid sequence of SEQ ID NO: 17 with respect to the HNRNPA2B1 gene:
(8) A gene mutation that causes at least one or more missense mutation, nonsense mutation, or frameshift mutation in the amino acid sequence of SEQ ID NO: 25 in the CSNK1A1 gene.
(9) For the CSNK2B gene, a gene mutation that causes at least one or more missense mutation, nonsense mutation, or frameshift mutation in the amino acid sequence of SEQ ID NO: 27:
(10) For the CSNK2A1 gene, a gene mutation that causes at least one or more nonsense mutation, frameshift mutation or splice site mutation in the amino acid sequence of SEQ ID NO: 29:
(11) A gene mutation that causes at least one or more mutations of the amino acid at position 124 and the amino acid at position 175 in the amino acid sequence of SEQ ID NO: 31 with respect to the CD28 gene.
13. 13. The method for testing a T-cell lymphoma according to any one of the above items 1 to 12, further comprising detecting an abnormal copy number of a gene in a sample collected from the subject.
14． About the specimen collected from the subject, the human chromosome 14q31.1 region, the human chromosome 7q31.1 region, the human chromosome 1p21.3 region of at least one or more chromosomes, at least one or more sites 14. The method for testing a T-cell lymphoma according to any one of the above items 1 to 13, which comprises detecting a deletion.
15. PRKCB gene, CCR4 gene, PLCG1 gene, CARD11 gene, CCR7 gene, STAT3 gene, TP53 gene, NOTCH1 gene, VAV1 gene, TBL1XR1 gene, GATA3 gene, IRF4 gene, FAS gene, POT1 gene, RHOA gene, IRF2BP2 gene, TET2 gene , HNRNPA2B1 gene, EP300 gene, CD58 gene, GPR183 gene, CSNK1A1 gene, CSNK2B gene, FYN gene, B2M gene, CBLB gene, CD28 gene, ZFP36L2 gene, CSNK2A1 gene, ATXN1 gene, SYNCRIP gene, S1PR1 gene, RELA gene, CDKN2A A method for screening a therapeutic agent for T-cell lymphoma, comprising screening for a gene and a substance capable of controlling a mutation or copy number abnormality of any one of the ICOS genes or a change in function of the gene due to the mutation or copy number abnormality.

  According to the test method of the present invention, T-cell lymphoma can be easily and effectively tested, and is useful information for diagnosing T-cell lymphoma, classifying T-cell lymphoma pathology and disease type, predicting the prognosis of T-cell lymphoma, and the like. Can be obtained. Furthermore, by confirming the copy number abnormality of the gene and the deletion of a specific region of the chromosome, information can be obtained, so that the test for T cell lymphoma can be performed with high accuracy. Further, according to the screening method of the present invention, a new therapeutic agent for T-cell lymphoma can be provided, and a therapeutic agent for a disease state with a poor prognosis can be screened.

  In the present invention, the whole picture of the genetic basis of T-cell lymphoma was clarified, and a plurality of genetic abnormalities that could serve as therapeutic targets and diagnostic markers for T-cell lymphoma could be identified. The present invention is based on the detection of gene mutations in a large number of cases to be analyzed as compared with the conventional report cases, and is considered to be highly clinically useful. Specifically, in the present invention, a new PLCG1 gene, PRKCB gene, CCR4 gene, CCR7 gene, CARD11 gene, RHOA gene, IRF4 gene, GATA3 gene, CD58 gene, POT1 gene, GPR183 gene, HNRNPA2B1 gene mutation, It was found that it was frequently observed. In particular, the PRKCB gene, CCR4 gene, CCR7 gene, GPR183 gene, and HNRNPA2B1 gene are genes for which no gene mutation has been reported in other tumors, and were considered to be mutations characteristic of T cell lymphoma. We have identified hot spots for mutations in the PRKCB, PLCG1, CCR4, CCR7, CARD11, RHOA, and IRF4 genes, and these genes are promising not only for diagnostic significance but also as therapeutic targets It is considered to be. In addition, in the present invention, it is possible to identify a characteristic gene mutation for each disease type, and it is considered to be useful for prognosis prediction. In the present invention, the molecular pathology of T-cell lymphoma has been comprehensively clarified, and a number of therapeutic targets for overcoming the pathology with a poor prognosis have been obtained.

FIG. 7 is a diagram showing the results of performing total exome analysis on specimens derived from normal tissues and tumor tissues of 38 patients. (Example 1 (1)) FIG. 4 is a diagram showing the results of identifying the types and frequencies of gene mutations by target sequence analysis for 355 cases including ATL and PTCL-NOS. (Example 1 (2)) It is a figure showing distribution of the gene mutation identified by target sequence analysis for every gene. (Example 1 (2)) It is a figure showing distribution of the gene mutation identified by target sequence analysis for every gene. (Example 1 (2)) It is a figure showing distribution of the gene mutation identified by target sequence analysis for every gene. (Example 1 (2)) It is a figure showing distribution of the gene mutation identified by target sequence analysis for every gene. (Example 1 (2)) It is a figure showing the frequency of the gene mutation identified by target sequence analysis. (Example 1 (2)) It is a figure which shows the result of having analyzed the mutation frequency for every ATL disease type. (Example 1 (2)), FIG. 4 is a view showing the result of analyzing a copy number abnormality of a gene by SNP array analysis. (Example 1 (3)) It is a figure showing the result of having analyzed the structural abnormality by whole genome analysis. (Example 1 (4)) FIG. 9 shows the results of Western blot confirming that mutant PRKCB (PRKCB D427N) enhances IKK phosphorylation and IκBα phosphorylation, and that PRKCB membrane translocation is enhanced. (Example 2 (1)) FIG. 9 shows the results of confirming that mutant PRKCB (PRKCB D427N) enhances NF-κB transcription activity by luciferase assay. (Example 2 (1)) FIG. 4 shows the results of examining the correlation between CCR4 expression and the presence or absence of a CCR4 mutation by flow cytometry. (Example 2 (2)). FIG. 2 shows that introduction of the CCR4 Y331X mutation increases expression of CCR4 on the membrane surface and suppresses receptor internalization by CCL22 ligand. (Example 2 (2))

  T cell lymphoma is a type of non-Hodgkin's lymphoma, a malignant tumor of the lymphatic system. Non-Hodgkin's lymphoma includes B-cell lymphoma and T-cell lymphoma. T-cell lymphoma includes adult T-cell leukemia / lymphoma (ATL), peripheral T-cell lymphoma, not otherwise specified: PTCL-NOS, vascular immunoblast Angioimmunoblastic T-cell lymphoma (AITL), Precursor T cell lymphoblastic lymphoma / leukemia (T-LBL), chronic T lymphocytic leukemia / progenitor lymphocytic Leukemia (T cell chronic lymphocytic leukemia / T-Prolymphocytic lymphoma: T-PLL), T-cell large granular lymphocyte leukemia (T-LGL), large-granular NK cell leukemia (NK-Cell) Large Granular Lymphocyte Leukemia (NK-LGL), Angiocentric Lymphoma, Intestinal T-Cell Lymphoma, Anaplastic large Cell (CD30 +) Lymphoma , Hodgkin / Hodgkin Seki Anaplastic large cell lymphoma (ALCL Hodgkin's-Like / Hodgkin's-related), include mycosis fungoides (Mycosis fungoides) and the like. The T-cell lymphoma according to the present invention is preferably ATL or PTCL-NOS.

  Adult T-cell leukemia / lymphoma (ATL) is known to have various symptoms and clinical courses. (1) Acute type (Acute), (2) Lymphoma type (Lymphoma) , (3) Chronic, and (4) Smoldering. (1) In the acute form, abnormal lymphocytes with nuclei in the form of petals appear in the blood and increase rapidly, and infections and elevated calcium levels in the blood may be seen. Requires immediate treatment with the drug. (2) In the lymphoma type, malignant lymphocytes proliferate mainly in the lymph nodes, and no abnormal cells are found in the blood. However, symptoms rapidly appear as in the acute type, and It is necessary to start treatment with cancer drugs. (3) In the chronic type, the number of white blood cells in the blood increases, and a large number of abnormal lymphocytes appear. However, the proliferation is not fast and many of them have almost no symptoms. An observation is made. (4) In the smoldering type, the white blood cell count is normal, but abnormal lymphocytes are present in the blood and may be accompanied by skin eruptions. Follow-up is performed. It is known that the acute type and the lymphoma type have a relatively poor prognosis, and the chronic type and the smoldering type have a relatively good prognosis.

  The test method of the present invention includes a step of detecting a gene mutation for at least one or more, preferably both, of two genes, PRKCB gene and CCR4 gene. In order to test T-cell lymphoma with higher accuracy, in addition to at least one of PRKCB gene and CCR4 gene, at least four of PLCG1, CARD11, CCR7, and STAT3 genes It is preferable that a step of detecting a gene mutation is included in at least one, preferably at least two, more preferably all genes. In order to test T cell lymphoma with higher precision, at least one of PRKCB gene and CCR4 gene and at least one of PLCG1, CARD11, CCR7, and STAT3 genes are required. In addition to the gene, TP53 gene, NOTCH1 gene, VAV1 gene, TBL1XR1 gene, GATA3 gene, IRF4 gene, FAS gene, POT1 gene, RHOA gene, IRF2BP2 gene, TET2 gene, HNRNPA2B1 gene, EP300 gene, CD58 gene, GPR183 gene, CSNK1A1 gene, CSNK2B gene, FYN gene, B2M gene, CBLB gene, CD28 gene, ZFP36L2 gene, CSNK2A1 gene, ATXN1 gene, SYNCRIP gene, S1PR1 gene, RELA gene, CDKN2A gene, and at least one of 29 genes of ICOS gene , Preferably at least two or more, more preferably at least five or more, even more preferably all genes. In addition, it is preferable that a step of detecting a gene mutation is included.

The sequence information for each of the above genes is as follows.

The PRKCB gene (Genbank Accession No. NM_002738: SEQ ID NO: 2) is a gene encoding a protein belonging to protein kinase C, which is a serine threonine-specific protein kinase. The PRKCB gene encodes the amino acid sequence shown in SEQ ID NO: 1 below.
SEQ ID NO: 1 (673aa): Genbank Accession No. NP_002729
1 MADPAAGPPP SEGEESTVRF ARKGALRQKN VHEVKNHKFT ARFFKQPTFC SHCTDFIWGF
61 GKQGFQCQVC CFVVHKRCHE FVTFSCPGAD KGPASDDPRS KHKFKIHTYS SPTFCDHCGS
121 LLYGLIHQGM KCDTCMMNVH KRCVMNVPSL CGTDHTERRG RIYIQAHIDR DVLIVLVRDA
181 KNLVPMDPNG LSDPYVKLKL IPDPKSESKQ KTKTIKCSLN PEWNETFRFQ LKESDKDRRL
241 SVEIWDWDLT SRNDFMGSLS FGISELQKAS VDGWFKLLSQ EEGEYFNVPV PPEGSEANEE
301 LRQKFERAKI SQGTKVPEEK TTNTVSKFDN NGNRDRMKLT DFNFLMVLGK GSFGKVMLSE
361 RKGTDELYAV KILKKDVVIQ DDDVECTMVE KRVLALPGKP PFLTQLHSCF QTMDRLYFVM
421 EYVNGGDLMY HIQQVGRFKE PHAVFYAAEI AIGLFFLQSK GIIYRDLKLD NVMLDSEGHI
481 KIADFGMCKE NIWDGVTTKT FCGTPDYIAP EIIAYQPYGK SVDWWAFGVL LYEMLAGQAP
541 FEGEDEDELF QSIMEHNVAY PKSMSKEAVA ICKGLMTKHP GKRLGCGPEG ERDIKEHAFF
601 RYIDWEKLER KEIQPPYKPK ACGRNAENFD RFFTRHPPVL TPPDQEVIRN IDQSEFEGFS
661 FVNSEFLKPE VKS
The mutation of the PRKCB gene associated with T cell lymphoma is at least one or more missense mutations, specifically, the amino acid at position 427, the amino acid at position 470, the amino acid at position 533 in the amino acid sequence of SEQ ID NO: 1, It is a gene mutation that results in a mutation of at least one amino acid at amino acid 546 and amino acid 630. More specifically, the gene mutation in the PRKCB gene is, specifically, substitution of aspartic acid at position 427 with asparagine, glycine or alanine (D427N, D427G, or D427A) in the amino acid sequence of SEQ ID NO: 1, amino acid at position 470 Substitution of histidine for histidine (D470H), substitution of glutamic acid at position 533 for lysine (E533K), substitution of glutamic acid at position 546 for valine (E546V), and substitution of aspartic acid at position 630 for asparagine, tyrosine, and glycine (D630N, D630Y, D630G).

The CCR4 gene (Genbank Accession No. NM_005508: SEQ ID NO: 4) is a gene encoding CC chemokine receptor 4 (CCR4). The CCR4 gene encodes the amino acid sequence shown in SEQ ID NO: 3 below.
SEQ ID NO: 3 (360aa): Genbank Accession No. NP_005499
1 MNPTDIADTT LDESIYSNYY LYESIPKPCT KEGIKAFGEL FLPPLYSLVF VFGLLGNSVV
61 VLVLFKYKRL RSMTDVYLLN LAISDLLFVF SLPFWGYYAA DQWVFGLGLC KMISWMYLVG
121 FYSGIFFVML MSIDRYLAIV HAVFSLRART LTYGVITSLA TWSVAVFASL PGFLFSTCYT
181 ERNHTYCKTK YSLNSTTWKV LSSLEINILG LVIPLGIMLF CYSMIIRTLQ HCKNEKKNKA
241 VKMIFAVVVL FLGFWTPYNI VLFLETLVEL EVLQDCTFER YLDYAIQATE TLAFVHCCLN
301 PIIYFFLGEK FRKYILQLFK TCRGLFVLCQ YCGLLQIYSA DTPSSSYTQS TMDHDLHDAL
The mutation of the CCR4 gene associated with T-cell lymphoma is at least one or more nonsense mutations or frameshift mutations in the C-terminal region, which is the amino acid sequence from 309 to 360 in the amino acid sequence of SEQ ID NO: 3. . More specifically, the gene mutation in the CCR4 gene is a frame shift mutation at the 322nd cysteine (C322fs) or 3 bases inserted at the position of the 322nd cysteine in the amino acid sequence of SEQ ID NO: 3; Arginine-induced mutation (C322delinsCR), frameshift at cysteine at position 323 (C323fs), conversion of glutamine at 330 to a stop codon (Q330X), conversion of tyrosine at position 331 to a stop codon (Y331X), position 336 Is a mutation that causes at least one or more conversions of glutamine to a stop codon (Q336X).

The PLCG1 (Phospholipase C, gamma 1) gene (Genbank Accession No. NM_002660: SEQ ID NO: 6) encodes the amino acid sequence shown in SEQ ID NO: 5 below.
Sequence number 5 (1291aa): Genbank Accession No. NP_002651
1 MAGAASPCAN GCGPGAPSDA EVLHLCRSLE VGTVMTLFYS KKSQRPERKT FQVKLETRQI
61 TWSRGADKIE GAIDIREIKE IRPGKTSRDF DRYQEDPAFR PDQSHCFVIL YGMEFRLKTL
121 SLQATSEDEV NMWIKGLTWL MEDTLQAPTP LQIERWLRKQ FYSVDRNRED RISAKDLKNM
181 LSQVNYRVPN MRFLRERLTD LEQRSGDITY GQFAQLYRSL MYSAQKTMDL PFLEASTLRA
241 GERPELCRVS LPEFQQFLLD YQGELWAVDR LQVQEFMLSF LRDPLREIEE PYFFLDEFVT
301 FLFSKENSVW NSQLDAVCPD TMNNPLSHYW ISSSHNTYLT GDQFSSESSL EAYARCLRMG
361 CRCIELDCWD GPDGMPVIYH GHTLTTKIKF SDVLHTIKEH AFVASEYPVI LSIEDHCSIA
421 QQRNMAQYFK KVLGDTLLTK PVEISADGLP SPNQLKRKIL IKHKKLAEGS AYEEVPTSMM
481 YSENDISNSI KNGILYLEDP VNHEWYPHYF VLTSSKIYYS EETSSDQGNE DEEEPKEVSS
541 STELHSNEKW FHGKLGAGRD GRHIAERLLT EYCIETGAPD GSFLVRESET FVGDYTLSFW
601 RNGKVQHCRI HSRQDAGTPK FFLTDNLVFD SLYDLITHYQ QVPLRCNEFE MRLSEPVPQT
661 NAHESKEWYH ASLTRAQAEH MLMRVPRDGA FLVRKRNEPN SYAISFRAEG KIKHCRVQQE
721 GQTVMLGNSE FDSLVDLISY YEKHPLYRKM KLRYPINEEA LEKIGTAEPD YGALYEGRNP
781 GFYVEANPMP TFKCAVKALF DYKAQREDEL TFIKSAIIQN VEKQEGGWWR GDYGGKKQLW
841 FPSNYVEEMV NPVALEPERE HLDENSPLGD LLRGVLDVPA CQIAIRPEGK NNRLFVFSIS
901 MASVAHWSLD VAADSQEELQ DWVKKIREVA QTADARLTEG KIMERRKKIA LELSELVVYC
961 RPVPFDEEKI GTERACYRDM SSFPETKAEK YVNKAKGKKF LQYNRLQLSR IYPKGQRLDS
1021 SNYDPLPMWI CGSQLVALNF QTPDKPMQMN QALFMTGRHC GYVLQPSTMR DEAFDPFDKS
1081 SLRGLEPCAI SIEVLGARHL PKNGRGIVCP FVEIEVAGAE YDSTKQKTEF VVDNGLNPVW
1141 PAKPFHFQIS NPEFAFLRFV VYEEDMFSDQ NFLAQATFPV KGLKTGYRAV PLKNNYSEDL
1201 ELASLLIKID IFPAKQENGD LSPFSGTSLR ERGSDASGQL FHGRAREGSF ESRYQQPFED
1261 FRISQEHLAD HFDSRERRAP RRTRVNGDNR L
The mutation of the PLCG1 gene associated with T cell lymphoma is at least one or more missense mutations, and is the amino acid at position 48, the amino acid at position 345, the amino acid at position 520, and the amino acid at position 748 in the amino acid sequence of SEQ ID NO: 5. , The amino acid at position 1016, the amino acid at position 1163, and the missense mutation of at least one amino acid at the amino acid at position 1165. More specifically, the gene mutation in the PLCG1 gene may be, for example, a substitution of arginine at position 48 with tryptophan (R48W), a substitution of serine at position 345 with phenylalanine (S345F), 520 in the amino acid sequence of SEQ ID NO: 5. Substitution of serine for phenylalanine at position (S520F), substitution of arginine at position 748 for cysteine or glycine (R748C, R748G), substitution of glutamine at position 1016 for leucine or arginine (Q1016L, Q1016R), position of glutamic acid at position 1163 (E1163K), and substitution of aspartic acid at position 1165 with histidine, glycine, or valine (D1165H, D1165G, D1165V).

The CCR7 (CC chemokine receptor 7) gene (Genbank Accession No. NM_001838: SEQ ID NO: 8) encodes the amino acid sequence shown in SEQ ID NO: 7 below.
SEQ ID NO: 7 (378aa): Genbank Accession No. NP_001829
1 MDLGKPMKSV LVVALLVIFQ VCLCQDEVTD DYIGDNTTVD YTLFESLCSK KDVRNFKAWF
61 LPIMYSIICF VGLLGNGLVV LTYIYFKRLK TMTDTYLLNL AVADILFLLT LPFWAYSAAK
121 SWVFGVHFCK LIFAIYKMSF FSGMLLLLCI SIDRYVAIVQ AVSAHRHRAR VLLISKLSCV
181 GIWILATVLS IPELLYSDLQ RSSSEQAMRC SLITEHVEAF ITIQVAQMVI GFLVPLLAMS
241 FCYLVIIRTL LQARNFERNK AIKVIIAVVV VFIVFQLPYN GVVLAQTVAN FNITSSTCEL
301 SKQLNIAYDV TYSLACVRCC VNPFLYAFIG VKFRNDLFKL FKDLGCLSQE QLRQWSSCRH
361 IRRSSMSVEA ETTTTFSP
The mutation of the CCR7 gene associated with T-cell lymphoma is at least one or more nonsense mutation or frameshift mutation in the C-terminal region which is the amino acid sequence from 332 to 378 in the amino acid sequence of SEQ ID NO: 7. More specifically, the mutation in the CCR7 gene is, in the amino acid sequence of SEQ ID NO: 7, the conversion of a glutamine at position 349 to a stop codon (Q349X), the conversion of a glutamine at position 351 to a stop codon (Q351X), The conversion of tryptophan at position 355 into a stop codon (W355X) results in at least one or more mutations.

CARD11 (Caspase recruitment domain-containing protein 11) gene (Genbank Accession No. NM_032415: SEQ ID NO: 10) encodes an amino acid sequence represented by the following SEQ ID NO: 9.
SEQ ID NO: 9 (1154aa): Genbank Accession No. NP_115791
1 MPGGGPEMDD YMETLKDEED ALWENVECNR HMLSRYINPA KLTPYLRQCK VIDEQDEDEV
61 LNAPMLPSKI NRAGRLLDIL HTKGQRGYVV FLESLEFYYP ELYKLVTGKE PTRRFSTIVV
121 EEGHEGLTHF LMNEVIKLQQ QMKAKDLQRC ELLARLRQLE DEKKQMTLTR VELLTFQERY
181 YKMKEERDSY NDELVKVKDD NYNLAMRYAQ LSEEKNMAVM RSRDLQLEID QLKHRLNKME
241 EECKLERNQS LKLKNDIENR PKKEQVLELE RENEMLKTKN QELQSIIQAG KRSLPDSDKA
301 ILDILEHDRK EALEDRQELV NRIYNLQEEA RQAEELRDKY LEEKEDLELK CSTLGKDCEM
361 YKHRMNTVML QLEEVERERD QAFHSRDEAQ TQYSQCLIEK DKYRKQIREL EEKNDEMRIE
421 MVRREACIVN LESKLRRLSK DSNNLDQSLP RNLPVTIISQ DFGDASPRTN GQEADDSSTS
481 EESPEDSKYF LPYHPPQRRM NLKGIQLQRA KSPISLKRTS DFQAKGHEEE GTDASPSSCG
541 SLPITNSFTK MQPPRSRSSI MSITAEPPGN DSIVRRYKED APHRSTVEED NDSGGFDALD
601 LDDDSHERYS FGPSSIHSSS SSHQSEGLDA YDLEQVNLMF RKFSLERPFR PSVTSVGHVR
661 GPGPSVQHTT LNGDSLTSQL TLLGGNARGS FVHSVKPGSL AEKAGLREGH QLLLLEGCIR
721 GERQSVPLDT CTKEEAHWTI QRCSGPVTLH YKVNHEGYRK LVKDMEDGLI TSGDSFYIRL
781 NLNISSQLDA CTMSLKCDDV VHVRDTMYQD RHEWLCARVD PFTDHDLDMG TIPSYSRAQQ
841 LLLVKLQRLM HRGSREEVDG THHTLRALRN TLQPEEALST SDPRVSPRLS RASFLFGQLL
901 QFVSRSENKY KRMNSNERVR IISGSPLGSL ARSSLDATKL LTEKQEELDP ESELGKNLSL
961 IPYSLVRAFY CERRRPVLFT PTVLAKTLVQ RLLNSGGAME FTICKSDIVT RDEFLRRQKT
1021 ETIIYSREKN PNAFECIAPA NIEAVAAKNK HCLLEAGIGC TRDLIKSNIY PIVLFIRVCE
1081 KNIKRFRKLL PRPETEEEFL RVCRLKEKEL EALPCLYATV EPDMWGSVEE LLRVVKDKIG
1141 EEQRKTIWVD EDQL
The mutation of the CARD11 gene associated with the T cell lymphoma is at least one or more missense mutations, and the amino acid at position 357, the amino acid at position 361, the amino acid at position 401, and the amino acid at position 615 in the amino acid sequence described in SEQ ID NO: 9. , And a gene mutation resulting in a mutation of at least one or more amino acids of the 626th amino acid. More specifically, the mutation in the CRAD11 gene is the substitution of aspartic acid at position 357 with asparagine or glycine (D357N, D357G), or the substitution of tyrosine at position 361 with cysteine or serine in the amino acid sequence of SEQ ID NO: 9. Substitution (Y361C, Y361S), substitution of aspartic acid at position 401 with asparagine or valine (D401N, D401V), substitution of serine at position 615 with phenylalanine (S615F), substitution of glutamic acid at position 626 with lysine (E626K) At least one of the following.

The GATA3 (GATA binding protein 3) gene (Genbank Accession No. NM_001002295: SEQ ID NO: 12) encodes the amino acid sequence represented by the following SEQ ID NO: 11.
SEQ ID NO: 11 (444aa): Genbank Accession No. NP_001002295
1 MEVTADQPRW VSHHHPAVLN GQHPDTHHPG LSHSYMDAAQ YPLPEEVDVL FNIDGQGNHV
61 PPYYGNSVRA TVQRYPPTHH GSQVCRPPLL HGSLPWLDGG KALGSHHTAS PWNLSPFSKT
121 SIHHGSPGPL SVYPPASSSS LSGGHASPHL FTFPPTPPKD VSPDPSLSTP GSAGSARQDE
181 KECLKYQVPL PDSMKLESSH SRGSMTALGG ASSSTHHPIT TYPPYVPEYS SGLFPPSSLL
241 GGSPTGFGCK SRPKARSSTE GRECVNCGAT STPLWRRDGT GHYLCNACGL YHKMNGQNRP
301 LIKPKRRLSA ARRAGTSCAN CQTTTTTLWR RNANGDPVCN ACGLYYKLHN INRPLTMKKE
361 GIQTRNRKMS SKSKKCKKVH DSLEDFPKNS SFNPAALSRH MSSLSHISPF SHSSHMLTTP
421 TPMHPPSSLS FGPHHPSSMV TAMG
The GATA3 gene mutation associated with T cell lymphoma is at least one or more nonsense mutation, frameshift mutation, and splice site mutation in the amino acid sequence of SEQ ID NO: 11. More specifically, the mutation of the GATA3 gene includes conversion of the 63rd tyrosine into a stop codon in the amino acid sequence of SEQ ID NO: 11 (Y63X), mutation of the splice site of exon 2 (mutation of the acceptor site or the donor site). ), Which results in at least one or more conversions of the 96th tryptophan to a stop codon (W96X).

The IRF4 (interferon regulatory factor 4) gene (Genbank Accession No. NM_001195286: SEQ ID NO: 14) encodes the amino acid sequence shown in SEQ ID NO: 13 below.
SEQ ID NO: 13 (450aa): Genbank Accession No. NP_001182215
1 MNLEGGGRGG EFGMSAVSCG NGKLRQWLID QIDSGKYPGL VWENEEKSIF RIPWKHAGKQ
61 DYNREEDAAL FKAWALFKGK FREGIDKPDP PTWKTRLRCA LNKSNDFEEL VERSQLDISD
121 PYKVYRIVPE GAKKGAKQLT LEDPQMSMSH PYTMTTPYPS LPAQVHNYMM PPLDRSWRDY
181 VPDQPHPEIP YQCPMTFGPR GHHWQGPACE NGCQVTGTFY ACAPPESQAP GVPTEPSIRS
241 AEALAFSDCR LHICLYYREI LVKELTTSSP EGCRISHGHT YDASNLDQVL FPYPEDNGQR
301 KNIEKLLSHL ERGVVLWMAP DGLYAKRLCQ SRIYWDGPLA LCNDRPNKLE RDQTCKLFDT
361 QQFLSELQAF AHHGRSLPRF QVTLCFGEEF PDPQRQRKLI TAHVEPLLAR QLYYFAQQNS
421 GHFLRGYDLP EHISNPEDYH RSIRHSSIQE
The mutation in the IRF4 gene associated with T cell lymphoma is at least one or more missense mutations, and is at least one of the amino acids at position 59, position 70, and position 114 in the amino acid sequence set forth in SEQ ID NO: 13. A genetic mutation that results in a mutation of one or more amino acids. More specifically, the mutations in the IRF4 gene are as follows: substitution of lysine at position 59 with arginine (K59R), substitution of leucine at position 70 with valine (L70V), Substitution of serine with arginine or asparagine (S114R, S114N).

The TBL1XR1 (transducin (beta) -like 1 X-linked receptor 1) gene (Genbank Accession No. NM_024665: SEQ ID NO: 16) encodes the amino acid sequence shown in SEQ ID NO: 15 below.
SEQ ID NO: 15 (514aa): Genbank Accession No. NP_078941
1 MSISSDEVNF LVYRYLQESG FSHSAFTFGI ESHISQSNIN GALVPPAALI SIIQKGLQYV
61 EAEVSINEDG TLFDGRPIES LSLIDAVMPD VVQTRQQAYR DKLAQQQAAA AAAAAAAASQ
121 QGSAKNGENT ANGEENGAHT IANNHTDMME VDGDVEIPPN KAVVLRGHES EVFICAWNPV
181 SDLLASGSGD STARIWNLSE NSTSGSTQLV LRHCIREGGQ DVPSNKDVTS LDWNSEGTLL
241 ATGSYDGFAR IWTKDGNLAS TLGQHKGPIF ALKWNKKGNF ILSAGVDKTT IIWDAHTGEA
301 KQQFPFHSAP ALDVDWQSNN TFASCSTDMC IHVCKLGQDR PIKTFQGHTN EVNAIKWDPT
361 GNLLASCSDD MTLKIWSMKQ DNCVHDLQAH NKEIYTIKWS PTGPGTNNPN ANLMLASASF
421 DSTVRLWDVD RGICIHTLTK HQEPVYSVAF SPDGRYLASG SFDKCVHIWN TQTGALVHSY
481 RGTGGIFEVC WNAAGDKVGA SASDGSVCVL DLRK
The mutation of the TBL1XR1 gene associated with T cell lymphoma is at least one or more missense mutation, nonsense mutation or frameshift mutation in the amino acid sequence set forth in SEQ ID NO: 15, and specifically 142 to 143, 230 , 290, 307, and 325 amino acids. More specifically, the mutation in the TBL1XR1 gene is a mutation (A142_N143delinsX) in which two bases are inserted into the amino acid sequence of SEQ ID NO: 15 at positions 142 to 143 to asparagine to generate a stop codon, and position 230. Substitution of serine for phenylalanine or proline, or deletion (S230F, S230P, S230del), substitution of threonine at position 290 with arginine or lysine (T290R, T290K), substitution of histidine at position 307 with arginine, proline, or tyrosine (H307R, H307P, H307Y), at least one of substitution of cysteine at position 325 with tyrosine or phenylalanine (C325Y, C325F).

HNRNPA2B1 (heterogeneous nuclear ribonucleoprotein A2 / B1) gene (Genbank Accession No. NM_002137: SEQ ID NO: 18) encodes the amino acid sequence represented by SEQ ID NO: 17 below.
SEQ ID NO: 17 (341aa): Genbank Accession No. NP_002128
1 MEREKEQFRK LFIGGLSFET TEESLRNYYE QWGKLTDCVV MRDPASKRSR GFGFVTFSSM
61 AEVDAAMAAR PHSIDGRVVE PKRAVAREES GKPGAHVTVK KLFVGGIKED TEEHHLRDYF
121 EEYGKIDTIE IITDRQSGKK RGFGFVTFDD HDPVDKIVLQ KYHTINGHNA EVRKALSRQE
181 MQEVQSSRSG RGGNFGFGDS RGGGGNFGPG PGSNFRGGSD GYGSGRGFGD GYNGYGGGPG
241 GGNFGGSPGY GGGRGGYGGG GPGYGNQGGG YGGGYDNYGG GNYGSGNYND FGNYNQQPSN
301 YGPMKSGNFG GSRNMGGPYG GGNYGPGGSG GSGGYGGRSR Y
The mutation of the HNRNPA2B1 gene associated with T cell lymphoma is at least one or more nonsense mutation, frameshift mutation, or splice site mutation in SEQ ID NO: 17, specifically, a splice site mutation of exon 10, and a sequence. It is a gene mutation that causes at least one mutation of the mutation at the 322nd amino acid in the amino acid sequence described in No. 17. The amino acid mutation at position 322 in the HNRNPA2B1 gene is more specifically a substitution of glycine at position 322 with arginine or a stop codon (G322R, G322X) in the amino acid sequence of SEQ ID NO: 17.

The VAV1 (vav 1 guanine nucleotide exchange factor) gene (Genbank Accession No. NM_005428: SEQ ID NO: 20) encodes the amino acid sequence represented by SEQ ID NO: 19 below.
SEQ ID NO: 19 (845aa): Genbank Accession No. NP_005419
1 MELWRQCTHW LIQCRVLPPS HRVTWDGAQV CELAQALRDG VLLCQLLNNL LPHAINLREV
61 NLRPQMSQFL CLKNIRTFLS TCCEKFGLKR SELFEAFDLF DVQDFGKVIY TLSALSWTPI
121 AQNRGIMPFP TEEESVGDED IYSGLSDQID DTVEEDEDLY DCVENEEAEG DEIYEDLMRS
181 EPVSMPPKMT EYDKRCCCLR EIQQTEEKYT DTLGSIQQHF LKPLQRFLKP QDIEIIFINI
241 EDLLRVHTHF LKEMKEALGT PGAANLYQVF IKYKERFLVY GRYCSQVESA SKHLDRVAAA
301 REDVQMKLEE CSQRANNGRF TLRDLLMVPM QRVLKYHLLL QELVKHTQEA MEKENLRLAL
361 DAMRDLAQCV NEVKRDNETL RQITNFQLSI ENLDQSLAHY GRPKIDGELK ITSVERRSKM
421 DRYAFLLDKA LLICKRRGDS YDLKDFVNLH SFQVRDDSSG DRDNKKWSHM FLLIEDQGAQ
481 GYELFFKTRE LKKKWMEQFE MAISNIYPEN ATANGHDFQM FSFEETTSCK ACQMLLRGTF
541 YQGYRCHRCR ASAHKECLGR VPPCGRHGQD FPGTMKKDKL HRRAQDKKRN ELGLPKMEVF
601 QEYYGLPPPP GAIGPFLRLN PGDIVELTKA EAEQNWWEGR NTSTNEIGWF PCNRVKPYVH
661 GPPQDLSVHL WYAGPMERAG AESILANRSD GTFLVRQRVK DAAEFAISIK YNVEVKHIKI
721 MTAEGLYRIT EKKAFRGLTE LVEFYQQNSL KDCFKSLDTT LQFPFKEPEK RTISRPAVGS
781 TKYFGTAKAR YDFCARDRSE LSLKEGDIIK ILNKKGQQGW WRGEIYGRVG WFPANYVEED
841 YSEYC
The mutation of the VAV1 gene associated with T cell lymphoma is at least one or more missense mutations, and the amino acids at positions 174, 175, 404, and 556 in the amino acid sequence of SEQ ID NO: 19. 798, the amino acid at position 798, and at least one amino acid at position 822. More specifically, the mutations in the VAV1 gene are as follows: substitution of the tyrosine at position 174 with cysteine (Y174C), substitution of glutamic acid at position 175 with valine (E175V), Substitution of lysine for arginine (K404R), substitution of glutamic acid at position 556 with aspartic acid or lysine (E556D, E556K), substitution of arginine at position 798 with proline or glutamine (R798P, R798Q), and substitution at position 822 It results in at least one or more substitutions of arginine for glutamine (R822Q).

The GPR183 (G protein-coupled receptor 183) gene (Genbank Accession No. NM_004951: SEQ ID NO: 22) encodes the amino acid sequence shown in SEQ ID NO: 21 below.
SEQ ID NO: 21 (361aa): Genbank Accession No. NP_004942
1 MDIQMANNFT PPSATPQGND CDLYAHHSTA RIVMPLHYSL VFIIGLVGNL LALVVIVQNR
61 KKINSTTLYS TNLVISDILF TTALPTRIAY YAMGFDWRIG DALCRITALV FYINTYAGVN
121 FMTCLSIDRF IAVVHPLRYN KIKRIEHAKG VCIFVWILVF AQTLPLLINP MSKQEAERIT
181 CMEYPNFEET KSLPWILLGA CFIGYVLPLI IILICYSQIC CKLFRTAKQN PLTEKSGVNK
241 KALNTIILII VVFVLCFTPY HVAIIQHMIK KLRFSNFLEC SQRHSFQISL HFTVCLMNFN
301 CCMDPFIYFF ACKGYKRKVM RMLKRQVSVS ISSAVKSAPE ENSREMTETQ MMIHSKSSNG
361 K
The mutation of the GPR183 gene associated with T cell lymphoma is at least one or more nonsense mutation, frameshift mutation or splice site mutation in the amino acid sequence of SEQ ID NO: 21.

The IRF2BP2 (interferon regulatory factor 2 binding protein 2) gene (Genbank Accession No. NM_001077397: SEQ ID NO: 24) encodes the amino acid sequence shown in SEQ ID NO: 23 below.
SEQ ID NO: 23 (571aa): Genbank Accession No. NP_001070865
1 MAAAVAVAAA SRRQSCYLCD LPRMPWAMIW DFTEPVCRGC VNYEGADRVE FVIETARQLK
61 RAHGCFPEGR SPPGAAASAA AKPPPLSAKD ILLQQQQQLG HGGPEAAPRA PQALERYPLA
121 AAAERPPRLG SDFGSSRPAA SLAQPPTPQP PPVNGILVPN GFSKLEEPPE LNRQSPNPRR
181 GHAVPPTLVP LMNGSATPLP TALGLGGRAA ASLAAVSGTA AASLGSAQPT DLGAHKRPAS
241 VSSSAAVEHE QREAAAKEKQ PPPPAHRGPA DSLSTAAGAA ELSAEGAGKS RGSGEQDWVN
301 RPKTVRDTLL ALHQHGHSGP FESKFKKEPA LTAVARTARK RKPSPEPEGE VGPPKINGEA
361 QPWLSTSTEG LKIPMTPTSS FVSPPPPTAS PHSNRTTPPE AAQNGQSPMA ALILVADNAG
421 GSHASKDANQ VHSTTRRNSN SPPSPSSMNQ RRLGPREVGG QGAGNTGGLE PVHPASLPDS
481 SLATSAPLCC TLCHERLEDT HFVQCPSVPS HKFCFPCSRQ SIKQQGASGE VYCPSGEKCP
541 LVGSNVPWAF MQGEIATILA GDVKVKKERD S
The mutation of the IRF2BP2 gene associated with T-cell lymphoma is a gene mutation that results in at least one or more missense mutations, nonsense mutations or frameshift mutations in the amino acid sequence of SEQ ID NO: 23. It is a mutation of the 194th amino acid. More specifically, the missense mutation in the IRF2BP2 gene is a substitution of glycine at position 194 with aspartic acid or valine (G194D, G194V) in the amino acid sequence of SEQ ID NO: 23.

The CSNK1A1 (casein kinase 1, alpha 1) gene (Genbank Accession No. NM_001892: SEQ ID NO: 26) encodes the amino acid sequence represented by SEQ ID NO: 25 below.
SEQ ID NO: 25 (337aa): Genbank Accession No. NP_001883
1 MASSSGSKAE FIVGGKYKLV RKIGSGSFGD IYLAINITNG EEVAVKLESQ KARHPQLLYE
61 SKLYKILQGG VGIPHIRWYG QEKDYNVLVM DLLGPSLEDL FNFCSRRFTM KTVLMLADQM
121 ISRIEYVHTK NFIHRDIKPD NFLMGIGRHC NKLFLIDFGL AKKYRDNRTR QHIPYREDKN
181 LTGTARYASI NAHLGIEQSR RDDMESLGYV LMYFNRTSLP WQGLKAATKK QKYEKISEKK
241 MSTPVEVLCK GFPAEFAMYL NYCRGLRFEE APDYMYLRQL FRILFRTLNH QYDYTFDWTM
301 LKQKAAQQAA SSSGQGQQAQ TPTGKQTDKT KSNMKGF
The mutation of the CSNK1A1 gene associated with T cell lymphoma is at least one or more missense mutations, and is at least one of the amino acids at 136, 160 and 198 in the amino acid sequence of SEQ ID NO: 25. It is a gene mutation that causes the above mutation. More specifically, the mutation in the CSNK1A1 gene is the substitution of 136th aspartic acid with histidine, glycine, or asparagine (D136H, D136G, D136N) in the amino acid sequence of SEQ ID NO: 25, or phenylalanine of the 160th leucine. (L160F), or substitution of glutamine at position 198 with lysine or glutamic acid (Q198K, Q198E).

The CSNK2B (casein kinase 2, beta polypeptide) gene (Genbank Accession No. NM_001320: SEQ ID NO: 28) encodes the amino acid sequence represented by SEQ ID NO: 27 below.
SEQ ID NO: 27 (215aa): Genbank Accession No. NP_001311
1 MSSSEEVSWI SWFCGLRGNE FFCEVDEDYI QDKFNLTGLN EQVPHYRQAL DMILDLEPDE
61 ELEDNPNQSD LIEQAAEMLY GLIHARYILT NRGIAQMLEK YQQGDFGYCP RVYCENQPML
121 PIGLSDIPGE AMVKLYCPKC MDVYTPKSSR HHHTDGAYFG TGFPHMLFMV HPEYRPKRPA
181 NQFVPRLYGF KIHPMAYQLQ LQAASNFKSP VKTIR
The mutation of the CSNK2B gene associated with T-cell lymphoma is at least one or more missense mutation, nonsense mutation, or frameshift mutation in the amino acid sequence of SEQ ID NO: 27, specifically, the 173rd amino acid, and It is a genetic mutation that results in a mutation of at least one of the 182nd amino acid. More specifically, the mutation in the CSNK2B gene is, for example, the conversion of the 173rd glutamic acid to a stop codon or the replacement with aspartic acid (E173X, E173D) and the 182nd glutamine in the amino acid sequence of SEQ ID NO: 27. It causes at least one mutation in codon conversion (Q182X).

The CSNK2A1 (casein kinase 2, alpha 1 polypeptide) gene (Genbank Accession No. NM_001895: SEQ ID NO: 30) encodes the amino acid sequence represented by SEQ ID NO: 29 below.
SEQ ID NO: 29 (391aa): Genbank Accession No. NP_001886
1 MSGPVPSRAR VYTDVNTHRP REYWDYESHV VEWGNQDDYQ LVRKLGRGKY SEVFEAINIT
61 NNEKVVVKIL KPVKKKKIKR EIKILENLRG GPNIITLADI VKDPVSRTPA LVFEHVNNTD
121 FKQLYQTLTD YDIRFYMYEI LKALDYCHSM GIMHRDVKPH NVMIDHEHRK LRLIDWGLAE
181 FYHPGQEYNV RVASRYFKGP ELLVDYQMYD YSLDMWSLGC MLASMIFRKE PFFHGHDNYD
241 QLVRIAKVLG TEDLYDYIDK YNIELDPRFN DILGRHSRKR WERFVHSENQ HLVSPEALDF
301 LDKLLRYDHQ SRLTAREAME HPYFYTVVKD QARMGSSSMP GGSTPVSSAN MMSGISSVPT
361 PSPLGPLAGS PVIAAANPLG MPVPAAAGAQ Q
The mutation of the CSNK2A1 gene associated with T cell lymphoma is at least one or more nonsense mutations or frameshift mutations in the amino acid sequence of SEQ ID NO: 29.

The CD28 gene (Genbank Accession No. NM_006139: SEQ ID NO: 32) encodes the amino acid sequence shown in SEQ ID NO: 31 below.
SEQ ID NO: 31 (220aa): Genbank Accession No. NP_006130
1 MLRLLLALNL FPSIQVTGNK ILVKQSPMLV AYDNAVNLSC KYSYNLFSRE FRASLHKGLD
61 SAVEVCVVYG NYSQQLQVYS KTGFNCDGKL GNESVTFYLQ NLYVNQTDIY FCKIEVMYPP
121 PYLDNEKSNG TIIHVKGKHL CPSPLFPGPS KPFWVLVVVG GVLACYSLLV TVAFIIFWVR
181 SKRSRLLHSD YMNMTPRRPG PTRKHYQPYA PPRDFAAYRS
The mutation of the CD28 gene associated with T cell lymphoma is at least one or more missense mutations, and is a mutation of at least one or more of the amino acid at position 124 and the amino acid at position 175 in the amino acid sequence described in SEQ ID NO: 31. More specifically, the mutation in the CD28 gene is a substitution of aspartic acid at position 124 with glutamic acid or valine (D124E, D124V), or a modification of threonine at position 175 with isoleucine or proline in the amino acid sequence of SEQ ID NO: 31. It causes at least one mutation of substitution (T175I, T175P).

  The sample used in the test method of the present invention is not particularly limited as long as it is a sample collected from a subject and can be expected to contain a nucleic acid or a protein derived from the gene for the various genes. Samples of the present invention include, for example, body fluids such as lymph nodes, tonsils, bone marrow, digestive organs, digestive organs, respiratory organs, spleen, liver, sensory organs, central nervous system, motor organs, skin, urine, lymph, and blood including peripheral blood. Selected. The nucleic acid relating to a gene means genomic DNA of the gene, mRNA which is a transcription product of the gene, and the like. The gene-derived protein means a protein molecule expressed from the gene, a degradation product thereof, and the like. The test method of the present invention can detect a gene mutation using a biological sample obtained by processing a sample, and the biological sample can be, for example, one prepared by extracting nucleic acids or proteins, and appropriately diluted. Can be used.

  In the test method of the present invention, a method for detecting a gene defect or mutation is not particularly limited, and is a known method used for detecting a gene mutation or polymorphism, for example, a base sequence determination method based on the Sanger method. Invader method, Taqman probe method, SMMD method (Simultaneous Multiple Mutation Detection System), PCR-RFLP method (polymerase chain reaction-restriction fragment length polymorphism), MASA method, base extension method, or a method using a next-generation sequencer, Modified methods and the like can be used. The invader method and the Taqman probe method have excellent detection sensitivity and accuracy, and can be detected using a probe or primer having a base sequence that specifically recognizes a mutation site. Probes and primers for detecting the deletion or mutation of the various genes can be designed and synthesized by a conventional method based on the various genes and their surrounding nucleotide sequences. When detecting gene mutations for a plurality of genes, the gene mutations may be detected for all the genes at the same time, or the gene mutations may be detected not at the same time but a plurality of times.

  The test method of the present invention can provide information for diagnosing T cell lymphoma and predicting the possibility of onset. When a gene mutation is detected for each of the above genes, the subject from which the sample has been collected can be determined to have T-cell lymphoma.

  The test method of the present invention can also provide information for identifying a type of T cell lymphoma. In particular, by detecting mutations in at least one of the STAT3 gene, TP53 gene, IRF4 gene and CD58 gene, ATL (1) acute type (Acute), (2) lymphoma type (Lymphoma), (3) It is possible to provide information for four classifications of chronic type (Chronic) and (4) smoldering type (Smoldering). The test method of the present invention can also provide information on the possibility of a disease exhibiting a condition similar to the types (1) to (4). Specifically, when a mutation is detected in the STAT3 gene, it is determined to be a (3) chronic type or (4) smoldering type of ATL or a disease exhibiting a disease state similar thereto, and the TP53 gene and the IRF4 gene If a mutation is detected in at least one or more of the CD58 genes, the disease may be determined to be ATL (1) acute type or (2) lymphoma type or a disease exhibiting a similar condition. it can. T-LGL is exemplified as a disease exhibiting a condition similar to (3) chronic type or (4) smoldering type of ATL. The test method of the present invention can identify a disease or disease type, and can select a treatment method according to the type of the disease or disease type.

  Further, the test method of the present invention can provide information for predicting the prognosis of T-cell lymphoma. In particular, if a mutation is detected in the STAT3 gene, the prognosis is judged to be good, and if a mutation is detected in at least one or more of the TP53 gene, IRF4 gene, and CD58 gene, the prognosis is poor. It can be determined that. Since the prognosis can be predicted by the test method of the present invention, a treatment method according to the prognosis can be selected.

  In the test method of the present invention, a T cell lymphoma test can be performed with higher accuracy by detecting an abnormal copy number of a gene in addition to the detection of the gene mutation. In diseases such as cancer, DNA fragments of various sizes ranging from several kilobase pairs to several megabase pairs in the human genome are known to increase or decrease during replication of cell division. I have. One or more genes are contained on the DNA fragment, and an increase or decrease in the copy number of the gene causes a change in function. In the present invention, the abnormal copy number of a gene is a concept including a case where the copy number is increased and a case where the copy number is decreased as compared with a normal value (for example, 2) of the copy number of the gene. Furthermore, in the present invention, the abnormal copy number of a gene includes a case where the copy number of the gene does not show an increase or decrease, but the heterozygosity of the chromosome is lost. Loss of heterozygosity means that when one normal gene and one abnormal gene are present at a specific locus in normal cells, the normal gene is lost and the function of the abnormal gene is exerted strongly. It can be done. That is, even if the copy number of a gene is normal, the absence of an allele and the presence of only a mutated gene are included in the gene copy number abnormality of the present invention.

  Measurement of the copy number of a gene can be performed using a standard method known in the art. By referring to the nucleotide sequence of each gene, the copy number of the gene can be measured using a standard method. Specific methods for measuring the copy number of a gene include analysis methods based on hybridization and amplification, such as Southern blotting and fluorescence in situ hybridization (FISH) (Angerer, Meth. Enzymol. 152, 649, 1987), a comparative genomic hybridization (CGH) method, a real-time quantitative PCR method, a SNP array method, a method using a next-generation sequencer, or the like.

  In the test method of the present invention, CD28 gene, CCR4 gene, DLG1 gene, IRF4 gene, CARD11 gene, CD247 gene, NOTCH1 gene, SMARCA4 gene, BCL11B gene, CCR7 gene, SPYD gene, CD58 gene, CD2 gene, IKZF2 gene, It is preferable to confirm an abnormality in the gene copy number of at least one of the TBL1XR1, TNFAIP3, TAX1BP1, LRRN3, CDKN2A, GATA3, ZEB1, GPR183, NRXN3, and TP53 genes.

  Further, in the test method of the present invention, human chromosome 14q31.1 region, human chromosome 7q31.1 region, human chromosome 1p21.3 region of at least one or more chromosome region, deletion of at least one or more sites. Preferably, detecting.

  Detection of the deletion of at least one or more sites on the chromosome can be performed using standard methods known in the art. By referring to the nucleotide sequence of each chromosome, a chromosomal site deletion can be detected using a standard method. Specific examples include analysis methods based on hybridization and amplification, such as Southern blotting, fluorescence in situ hybridization (FISH) (see Angerer, Meth. Enzymol. 152, 649, 1987), and comparative genomics. It can be detected by a method using a hybridization (CGH) method, a real-time quantitative PCR method, an SNP array method, a next-generation sequencer, or the like.

  The present invention also extends to a method for screening a therapeutic agent for T cell lymphoma. The screening method of the present invention is characterized by screening for a substance capable of controlling a gene mutation or copy number abnormality of any one of the above-mentioned genes possibly having various gene mutations or a change in the function of the gene caused by these mutations. The candidate compound to be selected in the screening method of the present invention may be any substance such as a low-molecular compound, a nucleic acid, and a protein. Hereinafter, the screening method will be described using the PRKCB gene as an example.

  The screening method of the present invention includes evaluating whether or not a candidate compound can affect the function of a PRKCB gene having a mutation (mutated PRKCB gene). Evaluation of whether a candidate compound can affect the function of a mutant PRKCB gene can be performed using cells expressing the mutant PRKCB encoded by the mutant PRKCB gene. Cells expressing the mutant PRKCB may be obtained by any method, and cells obtained by transformation by various known methods can be used. Evaluation of whether the candidate compound can affect the function of the mutant PRKCB gene depends on the expression level of the mutant PRKCB gene in the cells contacted with the candidate compound, the activity of the mutant PRKCB, the substance on which the mutant PRKCB acts Can be evaluated by measuring the amount / activity of the cell and the phenotype of the cell itself. For example, a mutant PRKCB having a D427N mutation enhances NF-κB transcription factor activity, so that when a candidate compound is brought into contact with cells expressing the mutant PRKCB, NF-κB transcription factor activity by the mutant PRKCB is increased. A substance that can suppress the enhancement of the gene can be screened by determining that the substance can control a change in the function of the gene.

  EXAMPLES The present invention will be described in more detail with reference to examples below for better understanding of the present invention, but it is needless to say that the present invention is not limited to these examples.

(Example 1) Identification of gene mutation and abnormal chromosomal structure in T-cell lymphoma In this example, patients diagnosed with ATL and PTCL-NOS among T-cell lymphomas and obtained informed consent were According to the protocol approved by the Ethics Committee of Kyoto University, a sample for the present invention was collected and comprehensive gene mutation analysis was performed.
In the present example, whole exome analysis was performed using samples derived from normal tissues and tumor tissues collected from 38 ATL patients, and comprehensive gene mutation analysis was performed. At the same time, multi-platform analysis was performed, including whole genome analysis (10 cases), transcriptome analysis (23 cases), methylome analysis (40 cases), and copy number analysis using SNP array (233 cases). In addition, follow-up sequences for identified gene mutations were performed using 355 T-cell lymphoma specimens. By combining these, we attempted to comprehensively elucidate the genetic abnormalities of T-cell lymphoma.

(1) Identification of Mutated Gene by Whole Exome Analysis First, whole exome analysis was performed on DNA from 38 normal ATL patient specimens derived from normal tissues and tumor tissues. Of the genomic region, the entire exon region encoding the protein (Exome) is concentrated using SureSelect Human All Exon V5 (Agilent Technology Co., Ltd.), which is a human all-exon capture kit, and then purified using HiSeq2000 (Illumina). The nucleotide sequence was determined. The mutation of the gene was confirmed by comparing the nucleotide sequence of the DNA derived from the tumor tissue with the nucleotide sequence of the DNA derived from the normal tissue.

  FIG. 1 shows the results of the whole exome analysis. A total of 2999 gene mutations (78.9 per case) were detected by whole exome analysis. When deep sequencing was performed to confirm the mutation of 332 genes in three cases, mutation was confirmed in 99.1% of all 332 genes. Next, as a result of analysis using MutSigCV, PLCG1 gene, PRKCB gene, CCR4 gene, CCR7 gene, CARD11 gene, RHOA gene, IRF4 gene, GATA3 gene, TP53 gene, FAS gene, CD58 gene, POT1 gene, GPR183 gene, Significantly more gene mutations were found in the HNRNPA2B1 gene.

(2) Identification of mutation frequency and analysis of mutation pattern by target sequence analysis
Target sequencing was performed using SureSelect Target Enrichment (Agilent Technology Co., Ltd.) on specimens derived from tumor tissues of 355 T-cell lymphoma patients, mainly ATL and PTCL-NOS patients. As target regions, genes identified by whole exome analysis and 146 genes belonging to pathways related thereto were targeted.

  The results of the target sequence analysis are shown in FIG. 2, FIGS. 3-1 to 3-4, and FIG. Of the genes found in the whole exome analysis, PLCG1 gene (37.2%), PRKCB gene (30.7%), CCR4 gene (25.9%), CCR7 gene (11.0%), CARD11 gene (19.7%), RHOA gene (11.0% ), IRF4 gene (13.0%), GATA3 gene (13.8%), TP53 gene (18.9%), FAS gene (12.4%), CD58 gene (5.9%), POT1 gene (9.9%), GPR183 gene (5.4%) In addition, HNRNPA2B1 gene (6.2%) was found to be frequently observed in follow-up cases. In particular, novel hot spot missense mutations (D427N, D630N) were confirmed in the PRKCB gene, and these were considered to be gain-of-function mutations. In the CCR4 gene and CCR7 gene, nonsense mutations and frameshift mutations were frequently detected in the C-terminal region. Furthermore, it is known that the PLCG1, CARD11, IRF4, RHOA, and GATA3 genes have mutations in other lymphoid tumors, but ATL shows a novel hot spot missense mutation that has not been reported previously. (PLCG1 gene: R48W, S345F, E1163K, D1165H; CARD11 gene: E626K; IRF4 gene: K59R, L70V; RHOA gene: C16R; GATA3 gene: exon2 splice site mutation) were observed. In addition to the PRKCB, CCR4, and CCR7 genes, the GPR183 gene and the HNRNPA2B1 gene have not been reported to have mutations in other malignant tumors. In addition to these, as genes that show significant abnormalities, STAT3 gene (20.0%), NOTCH1 gene (16.9%), VAV1 gene (15.8%), TBL1XR1 gene (15.5%), IRF2BP2 gene (7.0%), TET2 gene ( 9.3%), EP300 gene (5.9%), CSNK1A1 gene (5.1%), CSNK2B gene (4.5%), FYN gene (3.9%), B2M gene (4.2%), CBLB gene (4.5%), CD28 gene (3.1% %), ZFP36L2 gene (2.8%), CSNK2A1 gene (2.5%), ATXN1 gene (2.8%), SYNCRIP gene (2.5%), S1PR1 gene (2.3%), RELA gene (2.3%), CDKN2A gene (1.7% ) And the ICOS gene (1.4%).

  According to the present example, it was possible to clarify the overall picture of the T cell lymphoma gene abnormality. In particular, mutations characteristic of T-cell lymphoma could be detected in 94.6% of patients by searching for significantly high-frequency 35 genes such as PLCG1, PRKCB, and CCR4. In addition, mutations could be detected in 45.4% of patients by searching PRKCB gene and CCR4 gene. In addition, mutations could be detected in 63.9% of patients by searching for the top three genes (PLCG1, PRKCB, and CCR4 genes). Mutations could be detected in 73.0% of patients by searching for PLCG1, PRKCB, CCR4, CCR7, CARD11, and STAT3 genes. Mutations could be detected in 72.7% of patients by searching for PLCG1, PRKCB, CCR4 and TP53 genes. The results also indicate that the detection of these gene mutations can be applied to the diagnosis of T-cell lymphoma.

  Analysis of mutation frequency for each type of ATL revealed that in the acute and lymphoma types, the TP53 gene, CD58 gene, and IRF4 gene mutations were significantly higher, and in the chronic and smoldering types, the STAT3 gene mutation was significantly higher. (Fig. 5). This result suggests that these genes may be useful for identifying ATL types and predicting prognosis. Furthermore, it is known that gene mutations in the STAT3 gene are frequently found in T-cell large-granular lymphocytic leukemia, suggesting that chronic and smoldering forms of ATL may exhibit pathologies similar to this disease .

(4) Identification of copy number abnormality using SNP array
Copy number analysis was performed on SNP arrays using GeneChip Human Mapping 250K Nsp (Affymetrix) for samples derived from tumor tissues of 233 cases of initial ATL patients.

FIG. 6 shows the results of gene copy number analysis by SNP array. Copy number abnormalities were observed in many genes. Further analysis using GISTIC2.0 revealed that copy number amplification of CD28 gene, CD274 gene, IL2RA gene, TCRζ gene, BCL11B gene, etc., copy of CDKN2A gene, IKZF2 gene, TBL1XR1 gene, TNFAIP3 gene, BLIMP1 gene, etc. A number of deletions were identified. In addition, copy number abnormalities were observed in genes frequently mutated in T cell lymphoma, and copy number amplification was observed in CCR4 gene, IFR4 gene, NOTCH1 gene, CARD11 gene, GATA3 gene, CD58 gene, TP53 gene, GPR183 In the gene, copy number deletions were repeatedly observed. These genes were considered to be targets for both gene mutation and copy number abnormality.
The same analysis was performed using 41 tumor-derived specimens of PTCL-NOS patients, and a common copy number abnormality such as amplification of the CD28 gene copy number was confirmed.

(5) Identification of gene copy number abnormality and chromosome structure abnormality by whole genome analysis
Genome-wide DNA analysis was performed on samples from 10 normal ATL patients from normal and tumor tissues. In the whole genome analysis, the nucleotide sequence was determined using HiSeq2000 (Illumina).

  In the whole genome analysis, not only the gene mutation observed in the whole exome analysis was detected, but also new copy number abnormalities and chromosomal structural abnormalities were identified (FIG. 7). In particular, in the 14q31.1 region, a total of 38 (1-8 / case) focal deletions were observed in all cases (10/10 cases), and abnormalities in the 14q31.1 region characterize ATL genomic abnormalities. it was thought. In addition, in the 7q31.1 region, a total of 18 (1-5 / case) focal deletions were observed in 8/10 cases, and in the 1p21.3 region, a total of 10 (1-3 / case) focal deletions were observed in 6/10 cases. Was done. In addition, tandem duplication was observed in the CD28 gene (2/10 cases) and CD274 gene (3/10 cases), respectively, as an abnormality that was repeatedly observed. The tandem duplication of this CD28 gene was recognized as a fusion transcript in two cases in RNA sequencing. In addition, whole genome analysis was able to detect the HTLV-1 genome, and integration of 1 to 3 HTLV-1 genomes was identified in each case.

(Example 2) Functional analysis of identified gene mutations A plurality of gene mutations identified in Example 1 were subjected to functional analysis below.
(1) PRKCB D427N, in which the most hot-spot missense mutations were identified, showed that the structure of the NFD motif that anchors the C1B domain, which is important for membrane translocation, was destabilized by three-dimensional structural analysis. It was thought to be easy to activate. The characteristic of the molecular pathology of ATL is that NF-κB is activated, and protein kinase C containing PRKCB is known to function as an upstream of the NF-κB pathway. The effect on the pathway was evaluated by introducing a mutant PRKCB gene (D427 mutation) into 293T cells.

  Western blot revealed that PRKCB D427 enhanced phosphorylation of IKK (IκB kinase) and IκBα (FIG. 8). Furthermore, as a result of a luciferase assay, the mutated PRKCB significantly enhanced the NF-κB transcription activity as compared with the wild-type PRKCB (FIG. 9).

  Similarly, a similar experiment was performed using Jurkat cells, a T-cell leukemia strain, and it was found that the mutant PRKCB significantly increased NF-κB transcriptional activity under PMA / Ionomycin administration (FIG. 9). ).

  These results indicate that mutant PRKCB contributes to NF-κB pathway activation in ATL.

(2) It was examined whether or not there is a correlation between the expression of CCR4 and the presence or absence of a CCR4 mutation in the tumor cells of the first ATL patient. As a result, CCR4 expression was significantly increased in CCR4 mutation-positive T-cell lymphomatosis (FIG. 10).

  Furthermore, the mutant CCR4 gene (Y331X mutation) was introduced into Jurkat cells, and the expression of CCR4 on the membrane surface was evaluated by flow cytometry. When the mutant CCR gene was introduced, the expression of CCR4 was increased (Fig. 11). Furthermore, suppression of receptor internalization for CCL22 ligand was observed. These results suggest that the gene mutation in CCR4 may increase the expression of CCR4 on the membrane surface, suggesting that the presence or absence of the CCR4 gene mutation may correlate with the reactivity to anti-CCR4 antibody.

  According to the test method of the present invention, T-cell lymphoma can be easily and effectively tested, and useful information is obtained for diagnosis of T-cell lymphoma, classification of T-cell lymphoma pathology, prediction of T-cell lymphoma prognosis, and the like. be able to. Furthermore, by confirming the copy number abnormality of the gene and the deletion of a specific region of the chromosome, information can be obtained, so that the test for T cell lymphoma can be performed with high accuracy. Further, according to the screening method of the present invention, a new therapeutic agent for T-cell lymphoma can be provided, and a therapeutic agent for a disease state with a poor prognosis can be screened.

Claims (4)

ATL (adult T cell leukemia / ATL) comprising a step of detecting at least one or more mutations in the PRKCB gene and / or CCR4 gene in a sample collected from a subject, as described in 1) to 3) below. how to assist in the inspection of lymphoma):
1) a mutation in the PRKCB gene that results in a mutant PRKCB that enhances phosphorylation of IκB kinase;
2) a mutation in the PRKCB gene that results in a mutant PRKCB that increases NF-κB transcription activity;
3) A gene mutation in which a mutation in the CCR4 gene causes an increase in CCR4 expression. The mutation of the PRKCB gene according to 1) and / or 2) above is
The method according to claim 1, wherein the mutation is a gene mutation that results in substitution of aspartic acid at position 427 of the amino acid sequence of SEQ ID NO: 1 with asparagine (D427N) . The mutation of the CCR4 gene described in 3) above is
A CCR4 gene mutation that results convert (Y331X) to termination codon of the 331 th tyrosine in the amino acid sequence set forth in SEQ ID NO: 3, a method for assisting the inspection according to claim 1. The method according to any one of claims 1 to 3 , further comprising a step of detecting a mutation in both a PRKCB gene and a CCR4 gene in a sample collected from a subject.