JP7066140B2 - How to test for T-cell lymphoma - Google Patents

Description

The present invention relates to a method for examining 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 a Peripheral T-cell lymphoma (not otherwise specified: PTCL-NOS) or 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 have an extremely poor prognosis. The incubation period is long from HTLV-1 virus infection to the onset of ATL, and the protein Tax encoded by the HTLV-1 virus is inactivated at the time of ATL onset. From these facts, it is considered that the onset of ATL is associated with somatic mutations acquired after HTLV-1 virus infection. However, little is known about the genetic basis of T-cell lymphoma, including ATL.

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. Furthermore, ATL is diagnosed by confirming HLTV-1 integration by HTLV-1 antibody test and 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 method with many treatment-related deaths and is not indicated for the elderly and patients with complications. Treatment of ATL is selected from chemotherapy with anticancer drugs and hematopoietic stem cell transplantation. Recently, a new anti-CCR4 antibody drug against CC chemokine receptor 4 (CCR4) has been approved for the indication of CCR4-positive ATL. However, the current situation is that treatment methods for T-cell lymphoma centered on ATL are limited, and it is eagerly desired to understand the molecular pathology of T-cell lymphoma and develop pathologically specific treatment methods.

It has been reported that ATL can be diagnosed and predicted by measuring the methylation state 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 in PTCL-NOS and angioimmunoblastic T-cell lymphoma (AITL), the RHOA gene was obtained. 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 (Non-Patent Document 1: Nat Genet. 2014; 46). 2): 171-5, Non-Patent Document 3: Nat Genet. 2014; 46 (4): 371-5). In addition, as a genetic analysis of ATL, the 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 USA. 2010; 107 (38): 1669-24.), JAK3 gene (Non-Patent Document 7: Blood. 2011) Oct 6; 118 (14): 3911-21) gene mutation, CDKN2A gene (Non-Patent Document 8: J Clin Oncol. 1997; 15 (5): 1778-85) and ZEB1 by gene copy number analysis 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) It has been reported that there is a deletion of 2012; 21 (1): 121-35) and amplification of the CARD11 gene and BCL11B gene (Non-Patent Document 12: Blood. 2006; 107 (11): 4500-7). There is.

In the above-mentioned reported cases, genetic analysis was performed on a small number of cases of PTCL-NOS and AITL, and although genetic abnormalities dependent on individual cases were detected, the overall picture of genetic abnormalities in T-cell lymphoma remains unclear. Is. In addition, many of the identified genetic abnormalities remain unclear in their functional significance and have not led to understanding of molecular pathologies or the development of pathologically specific therapies. Comprehensive genetic analysis using a large number of cases is desired to clarify the overall picture of gene abnormalities in T-cell lymphoma, but it has not been realized.

Japanese Unexamined Patent Publication No. 2007-244377 Japanese Unexamined Patent Publication No. 2009-254357

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 clarify the molecular pathology of T-cell lymphoma and to provide a new test method and therapeutic target for T-cell lymphoma.

In order to solve the above problems, the present inventors comprehensively analyzed gene abnormalities that occur frequently using T-cell lymphoma specimens in more than 350 cases. Focusing on the fact that high-frequency gene mutations are detected in 35 types of genes including the PRKCB gene and CCR4 gene, in particular, by detecting gene mutations in at least one or more of the PRKCB gene and CCR4 gene. , Have found that it is possible to test for T-cell lymphoma, and completed the present invention.

That is, the present invention comprises the following.
1. 1. For T-cell lymphoma , including the step of detecting mutations in at least one of the PLCG1 gene, CARD11 gene, CCR7 gene, VAV1 gene, GATA3 gene, IRF4 gene and HNRNPA2B1 gene in a sample collected from a subject. How to assist the inspection.
2. 2. The method for assisting the examination of T-cell lymphoma according to the preceding item 1, wherein the mutation of each gene is the gene mutation shown in the following (1) to (7):
(1) A mutation in the PLCG1 gene is at least one selected from the amino acids 48th, 345th, 520th, 748th, 1016th, 1163th, and 1165th in the amino acid sequence set forth in SEQ ID NO: 5. Missense mutations in genes that result in the above amino acid mutations;
(2) Mutations in the CARD11 gene include mutations in at least one amino acid selected from the amino acids 357, 361, 401, 615, and 626 in the amino acid sequence set forth in SEQ ID NO: 9. Missense mutations in the resulting gene;
(3) A nonsense or frameshift mutation in a gene in which a mutation in the CCR7 gene results in a mutation in at least one or more amino acids in the C-terminal region at positions 332-378 of the amino acid sequence set forth in SEQ ID NO: 7.
(4) A mutation in the VAV1 gene is at least one amino acid selected from the amino acids 174th, 175th, 404th, 556th, 798th, and 822th in the amino acid sequence set forth in SEQ ID NO: 19. Missense mutations in genes that result in mutations in
(5) Mutations in the GATA3 gene include at least one mutation selected from the conversion of amino acids 63 and 96 in the amino acid sequence set forth in SEQ ID NO: 11 to stop codons and the splice site mutation of exon 2. Gene mutations that result;
(6) A missense mutation in a gene resulting in a mutation in at least one amino acid selected from the 59th, 70th, and 114th amino acids in the amino acid sequence set forth in SEQ ID NO: 13;
(7) At least one gene mutation of the HNRNPA2B1 gene mutation resulting in a splice site mutation of Exxon 10 in the amino acid sequence set forth in SEQ ID NO: 17 and a gene mutation resulting in a mutation of the 322nd amino acid. ..
3. 3. Modifications of the PLCG1 gene in the amino acid sequence set forth in SEQ ID NO: 5 include the substitution of arginine at position 48 with tryptophan (R48W), the substitution of serine at position 345 with phenylalanine (S345F), and the substitution of serine at position 520 with phenylalanine. Substitution (S520F), Substitution of 748th arginine to cysteine (R748C), Substitution of 748th arginine to glycine (R748G), Substitution of 1016th glutamine to leucine (Q1016L), Substitution of 1016th glutamine to arginine Substitution to (Q1016R), replacement of glutamic acid at position 1163 with lysine (E1163K), replacement of aspartic acid at position 1165 with histidine (D1165H), replacement of aspartic acid at position 1165 with glycine (D1165G), and 1165. The method for assisting the examination of T-cell lymphoma according to the preceding paragraph 1 or 2, which is a gene mutation resulting in at least one mutation selected from the substitution of the second aspartic acid with valine (D1165V) .
4. The mutation in the CARD11 gene is the substitution of aspartic acid at position 357 with aspartin (D357N), the substitution of aspartic acid at position 357 with lysine (D357G), and the cysteine at position 361 in the amino acid sequence set forth in SEQ ID NO: 9. Substitution to (Y361C), Substitution of 361st tyrosine to serine (Y361S), Substitution of 401st aspartic acid to aspartic acid (D401N), Substitution of 401st aspartic acid to valine (D401V), 615th The T-cell lymphoma according to item 1 or 2 above, which is a gene mutation that results in at least one mutation selected from the substitution of serine with phenylalanine (S615F) and the substitution of glutamic acid at position 626 with lysine (E626K). How to assist in the inspection of.
5. Mutations in the CCR7 gene include the conversion of glutamine at position 349 to a stop codon (Q349X), the conversion of glutamine at position 351 to a stop codon (Q351X), and the conversion of tryptophan at position 355 in the amino acid sequence set forth in SEQ ID NO: 7. The method for assisting the examination of T-cell lymphoma according to the preceding item 1 or 2, which is a gene mutation that results in at least one mutation selected from conversion to a stop codon (W355X).
6. Mutations in the VAV1 gene in the amino acid sequence set forth in SEQ ID NO: 19 include the substitution of tyrosine at position 174 with cysteine (Y174C), the substitution of glutamic acid at position 175 with valine (E175V), and the substitution of lysine at position 404 with arginine. Substitution (K404R), Glutamic acid at position 556 with alparaginic acid (E556D), Glutamic acid at position 556 with lysine (E556K), Arginine at position 798 with proline (R798P), Arginine at position 798 The test for T-cell lymphoma according to item 1 or 2 above, which is a gene mutation that results in at least one mutation selected from the substitution with glutamine (R798Q) and the substitution of arginine at position 822 with glutamine (R822Q). How to assist.
7. Mutations in the GATA3 gene include the conversion of tyrosine at position 63 to the stop codon (Y63X) in the amino acid sequence set forth in SEQ ID NO: 11, the mutation at the splice site of Exxon 2, and the conversion of tryptophan at position 96 to the stop codon (W96X). The method for assisting the examination of T-cell lymphoma according to the above item 1 or 2, which is a gene mutation that causes at least one mutation selected from).
8. Mutations in the IRF4 gene include the substitution of lysine at position 59 with arginine (K59R), the substitution of leucine at position 70 with valine (L70V), and the substitution of serine at position 114 with arginine in the amino acid sequence set forth in SEQ ID NO: 13. Assists in the examination of T-cell lymphoma according to item 1 or 2 above, which is a gene mutation that results in at least one mutation selected from the substitution (S114R) and the substitution of serine at position 114 with asparaginine (S114N). Method.
9. Mutations in the HNRNPA2B1 gene are the substitution of glycin at position 322 with arginine (G322R), the conversion of glycine at position 322 to the stop codon (G322X) and the splice site of Exxon 10 in the amino acid sequence set forth in SEQ ID NO: 17. The method for assisting the examination of T-cell lymphoma according to the preceding item 1 or 2, which is a gene mutation that results in at least one mutation selected from the mutations.
10. The T-cell lymphoma test according to any one of 1 to 3 above, which comprises a step of detecting each gene mutation of the PRKCB gene and the CCR4 gene in addition to the mutation of the PLCG1 gene in the sample collected from the subject. How to assist.
11. The method for assisting the examination of T-cell lymphoma according to the above item 10, wherein the mutation of the PRKCB gene is the following a) or b) and the mutation of the CCR4 gene is the gene mutation shown in c):
a) A gene mutation in which a mutation in the PRKCB gene results in a mutant PRKCB that enhances phosphorylation of IκB kinase;
b) Mutations in the PRKCB gene result in mutant PRKCBs that increase NF-κB transcriptional activity;
c) A gene mutation in which a mutation in the CCR4 gene results in increased expression of CCR4.

The test method of the present invention can easily and effectively test T-cell lymphoma, and is useful information for diagnosis of T-cell lymphoma, classification of pathophysiology and type of T-cell lymphoma, prediction of prognosis of T-cell lymphoma, and the like. Can be obtained. Furthermore, by confirming abnormal gene copy counts and deletions of specific regions of chromosomes and obtaining information, T-cell lymphoma can be examined 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 pathological condition with a poor prognosis can also be screened.

In the present invention, the overall picture of the genetic basis of T-cell lymphoma was clarified, and a plurality of genetic abnormalities that could be 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 conventionally reported cases, and is considered to be highly clinically useful. Specifically, in the present invention, the gene mutations of the PLCG1 gene, PRKCB gene, CCR4 gene, CCR7 gene, CARD11 gene, RHOA gene, IRF4 gene, GATA3 gene, CD58 gene, POT1 gene, GPR183 gene, and HNRNPA2B1 gene are newly introduced. We 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 mutations have been reported in other tumors, and are considered to be characteristic mutations in T-cell lymphoma. In addition, hotspots of gene mutations have been identified in the PRKCB gene, PLCG1 gene, CCR4 gene, CCR7 gene, CARD11 gene, RHOA gene, and IRF4 gene, and these genes are promising not only for their diagnostic significance but also as therapeutic targets. Is considered to be. In addition, in the present invention, a gene mutation characteristic of each disease type can be identified, which is considered to be useful for prognosis prediction. In the present invention, the molecular pathological condition of T-cell lymphoma has been comprehensively clarified, and a large number of therapeutic targets for overcoming the pathological condition having a poor prognosis could be obtained.

It is a figure which shows the result of having performed the whole exome analysis about the sample derived from the normal tissue and the tumor tissue of 38 patients. (Example 1 (1)) It is a figure which shows the result of having identified the type and frequency of a gene mutation by the target sequence analysis of 355 cases including ATL and PTCL-NOS. (Example 1 (2)) It is the figure which showed the distribution of the gene mutation identified by the target sequence analysis for each gene. (Example 1 (2)) It is the figure which showed the distribution of the gene mutation identified by the target sequence analysis for each gene. (Example 1 (2)) It is the figure which showed the distribution of the gene mutation identified by the target sequence analysis for each gene. (Example 1 (2)) It is the figure which showed the distribution of the gene mutation identified by the target sequence analysis for each gene. (Example 1 (2)) It is a figure which showed the frequency of the gene mutation identified by the target sequence analysis. (Example 1 (2)) It is a figure which shows the result of having analyzed the mutation frequency for each type of ATL. (Example 1 (2)), It is a figure which shows the result of having analyzed the copy number abnormality of a gene by SNP array analysis. (Example 1 (3)) It is a figure which shows the result of having analyzed the structural abnormality by whole genome analysis. (Example 1 (4)) It is a figure which shows the result of having confirmed that the mutant PRKCB (PRKCB D427N) promotes the phosphorylation of IKK and the phosphorylation of IκBα by Western blotting, and the membrane metastasis of PRKCB is promoted. (Example 2 (1)) It is a figure which shows the result of having confirmed that the mutant PRKCB (PRKCB D427N) enhances the NF-κB transcription activity by the luciferase assay. (Example 2 (1)) It is a figure which shows the result of having examined the correlation between the expression of CCR4 and the presence or absence of a CCR4 mutation by flow cytometry. (Example 2 (2)). It is a figure showing that the expression of CCR4 on the membrane surface is increased and the internalization of the receptor by the CCL22 ligand is suppressed by introducing the CCR4 Y331X mutation. (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), unclassifiable peripheral T-cell lymphoma (not otherwise specified: PTCL-NOS), and vascular immunoblasts. Angioimmunoblastic T-cell lymphoma (AITL), Precursor T cell lymphoblastic lymphoma / leukemia (T-LBL), Chronic T-lymphoma / precursor lymphoma Leukemia (T cell chronic lymphocytic leukemia / T-Prolymphocytic lymphoma: T-PLL), T-Cell Large Granular Lymphocyte Leukemia (T-LGL), Large granule NK cell leukemia (NK-Cell) Large Granular Lymphocyte Leukemia: NK-LGL, Angiocentric Lymphoma, Intestinal T-Cell Lymphoma, Anaplastic large Cell (CD30 +) Lymphoma , Hodgkin-like / Hodgkin-related undifferentiated large cell lymphoma (ALCL Hodgkin's-Like / Hodgkin's-related), mycosis fungoides, etc. The T-cell lymphoma in the present invention is preferably ATL or PTCL-NOS.

Adult T-cell leukemia / lymphoma (ATL) is known to have diverse symptoms and clinical course, (1) acute type (Acute), (2) lymphoma type (Lymphoma). , (3) Chronic type (4) Smoldering type (Smoldering). (1) In the acute type, abnormal lymphocytes with petal-shaped nuclei appear in the blood and increase rapidly, and infectious diseases and increased calcium levels in the blood may be seen, and anticancer cancer. 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. Treatment with lymph should be started. (3) In the chronic type, the number of white blood cells in the blood increases and a large number of abnormal lymphocytes appear, but their proliferation is not fast and many of them have almost no symptoms. Observations are made. (4) The smoldering type has a normal white blood cell count, but abnormal lymphocytes are present in the blood and may be accompanied by a rash. In many cases, it does not change for a long period of time without treatment. Follow-up is done. It is known that the acute type and 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 in at least one or more of two genes, a PRKCB gene and a CCR4 gene, preferably both genes. For more accurate testing of T-cell lymphoma, at least one or more of the two PRKCB and CCR4 genes, as well as at least four of the PLCG1, CARD11, CCR7, and STAT3 genes. It is preferable to include a step of detecting a gene mutation in one or more, preferably at least two or more, more preferably all genes. For more accurate testing of T-cell lymphoma, at least one of two genes, PRKCB gene and CCR4 gene, and at least one of four genes, PLCG1 gene, CARD11 gene, CCR7 gene, and STAT3 gene. In addition to genes, 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, At least one of 29 genes: 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. It is preferable to include a step of detecting a gene mutation, preferably at least two or more, more preferably at least five or more, and further preferably all genes.

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
Mutations in the PRKCB gene associated with T-cell lymphoma are at least one missense mutation, specifically amino acids 427, 470, 533, in the amino acid sequence set forth in SEQ ID NO: 1. A genetic mutation that results in a mutation in at least one or more amino acids at amino acid 546 and 630. More specifically, the gene mutation in the PRKCB gene is the substitution of aspartic acid at position 427 with aspartic acid, glycine or alanine (D427N, D427G, or D427A) in the amino acid sequence set forth in SEQ ID NO: 1, amino acid at position 470. Substitution with histidine (D470H), substitution of glutamic acid at position 533 with lysine (E533K), substitution of glutamic acid at position 546 with valine (E546V), and substitution of aspartic acid at position 630 with aspartic acid, tyrosine, glycine. A mutation that results in at least one or more of (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
Mutations in the CCR4 gene associated with T-cell lymphoma are at least one nonsense or frameshift mutation in the C-terminal region of the amino acid sequence 309-360 in the amino acid sequence set forth in SEQ ID NO: 3. .. More specifically, the gene mutation in the CCR4 gene is a frameshift mutation (C322fs) at the 322nd cysteine or 3 bases inserted at the position of the 322nd cysteine in the amino acid sequence set forth in SEQ ID NO: 3, and the cysteine and Mutation that produces arginine (C322delinsCR), frameshift at position 323 (C323fs), conversion of glutamine at position 330 to stop codon (Q330X), conversion of tyrosine at position 331 to stop codon (Y331X), position 336 A mutation that results in at least one or more of the conversion 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.
SEQ ID NO: 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 DEEEP KEVSS
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 PLCG1 gene mutation associated with T-cell lymphoma is at least one missense mutation, the 48th amino acid, the 345th amino acid, the 520th amino acid, and the 748th amino acid in the amino acid sequence set forth in SEQ ID NO: 5. , 1016th amino acid, 1163th amino acid, and 1165th amino acid, which is a missense variation of at least one amino acid. More specifically, the gene mutation in the PLCG1 gene is the substitution of arginine at position 48 with tryptophan (R48W), the substitution of serine at position 345 with phenylalanine (S345F), 520 in the amino acid sequence set forth in SEQ ID NO: 5. Substitution of the second serine with phenylalanine (S520F), substitution of the 748th arginine with cysteine or glycine (R748C, R748G), substitution of the 1016th glutamine with leucine or arginine (Q1016L, Q1016R), 1163th glutamic acid. It results in at least one or more substitutions for lysine (E1163K), histidine, glycine, or valine at position 1165 (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
Mutations in the CCR7 gene associated with T-cell lymphoma are at least one nonsense or frameshift mutation in the C-terminal region, which is the amino acid sequence from position 332 to 378 in the amino acid sequence set forth in SEQ ID NO: 7. More specifically, the mutation in the CCR7 gene includes the conversion of glutamine at position 349 to a stop codon (Q349X) and the conversion of glutamine at position 351 to a stop codon (Q351X) in the amino acid sequence set forth in SEQ ID NO: 7. It results in at least one mutation in the conversion of tryptophan at position 355 to a stop codon (W355X).

The CARD11 (Caspase recruitment domain-containing protein 11) gene (Genbank Accession No. NM_032415: SEQ ID NO: 10) encodes the amino acid sequence shown in SEQ ID NO: 9 below.
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 in the CARD 11 gene associated with T-cell lymphoma is at least one missense mutation, amino acid 357, 361, 401, 615 in the amino acid sequence set forth in SEQ ID NO: 9. , And a gene mutation that results in a mutation in at least one or more amino acids of amino acid 626. More specifically, the mutation in the CRAD11 gene is the substitution of aspartic acid at position 357 with aspartic acid or glycine (D357N, D357G) in the amino acid sequence set forth in SEQ ID NO: 9, and the mutation with tyrosine at position 361 to cysteine or serine. Substitution (Y361C, Y361S), substitution of aspartic acid at position 401 with aspartic acid or valine (D401N, D401V), substitution of serine at position 615 with phenylalanine (S615F), substitution of glutamic acid at position 626 with lysine (E626K) It brings at least one or more of.

The GATA3 (GATA binding protein 3) gene (Genbank Accession No. NM_001002295: SEQ ID NO: 12) encodes the amino acid sequence shown in SEQ ID NO: 11 below.
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
Mutations in the GATA3 gene associated with T-cell lymphoma are at least one nonsense mutation, frameshift mutation, or splice site mutation in the amino acid sequence set forth in SEQ ID NO: 11. More specifically, mutations in the GATA3 gene include conversion of the 63rd tyrosine to the stop codon (Y63X) in the amino acid sequence set forth in SEQ ID NO: 11, mutations in the splice site of Exxon 2 (mutations in the acceptor site or donor site). ), Which results in at least one or more conversions (W96X) of the 96th tryptophan to the stop codon.

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 VERSQL DISD
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
Mutations in the IRF4 gene associated with T-cell lymphoma are at least one missense mutation, at least the amino acids 59, 70, and 114 of the amino acid sequence set forth in SEQ ID NO: 13. A genetic mutation that results in a mutation in one or more amino acids. More specifically, the mutations in the IRF4 gene include the substitution of lysine at position 59 with arginine (K59R), the substitution of leucine at position 70 with valine (L70V), and position 114 in the amino acid sequence set forth in SEQ ID NO: 13. It results in at least one mutation of the 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 set forth 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 IIWD AHTGEA
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
Mutations in the TBL1XR1 gene associated with T-cell lymphoma are at least one missense mutation, nonsense mutation or frameshift mutation in the amino acid sequence set forth in SEQ ID NO: 15, specifically positions 142-143 and 230. , 290th, 307th, and 325th amino acids are genetic mutations that result in mutations in at least one or more amino acids. More specifically, the mutation in the TBL1XR1 gene is a mutation (A142_N143delinsX) in the amino acid sequence set forth in SEQ ID NO: 15, in which two bases are inserted at the positions of the 142nd alanine to the 143rd asparagine to generate a termination codon (A142_N143delinsX), the 230th. Replacement or deletion of serine with phenylalanine or proline (S230F, S230P, S230del), substitution of 290th threonine with arginine or lysine (T290R, T290K), 307th histidine to arginine, proline, or tyrosine (H307R, H307P, H307Y), at least one of the substitutions of cysteine at position 325 with tyrosine or phenylalanine (C325Y, C325F).

The HNRNPA2B1 (heterogeneous nuclear ribonucleoprotein A2 / B1) gene (Genbank Accession No. NM_002137: SEQ ID NO: 18) encodes the amino acid sequence set forth in 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
Mutations in the HNRNPA2B1 gene associated with T-cell lymphoma are at least one nonsense mutation, frameshift mutation, or splice site mutation in SEQ ID NO: 17, specifically the splice site mutation and sequence of Exxon 10. A gene mutation that results in at least one mutation in the mutation at amino acid 322 in the amino acid sequence set forth in number 17. The 322nd amino acid mutation in the HNRNPA2B1 gene is more specifically the substitution of 322nd glycine with an arginine or stop codon (G322R, G322X) in the amino acid sequence set forth in 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 shown in 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 in the VAV1 gene associated with T-cell lymphoma is at least one missense mutation, amino acid 174th, 175th, 404th, 556th in the amino acid sequence set forth in SEQ ID NO: 19. , 798th amino acid, and 822th amino acid is a genetic mutation that results in mutations in at least one or more amino acids. More specifically, the mutations in the VAV1 gene include the substitution of tyrosine at position 174 with cysteine (Y174C), the substitution of glutamic acid at position 175 with arginine (E175V), and position 404 in the amino acid sequence set forth in SEQ ID NO: 19. Replacement of lysine with arginine (K404R), replacement of glutamic acid at position 556 with arginine or lysine (E556D, E556K), replacement of arginine at position 798 with proline or glutamine (R798P, R798Q), and position 822. It results in at least one or more substitutions of arginine with 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
Mutations in the GPR183 gene associated with T-cell lymphoma are at least one nonsense mutation, frameshift mutation or splice site mutation in the amino acid sequence set forth in 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 set forth 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
Mutations in the IRF2BP2 gene associated with T-cell lymphoma are gene mutations that result in at least one or more missense, nonsense or frameshift mutations in the amino acid sequence set forth in SEQ ID NO: 23, wherein the missense mutation is specifically. It is a mutation of the 194th amino acid. The missense mutation in the IRF2BP2 gene is more specifically the substitution of glycine at position 194 with aspartic acid or valine (G194D, G194V) in the amino acid sequence set forth in 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 set forth in 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
Mutations in the CSNK1A1 gene associated with T-cell lymphoma are at least one missense mutation and at least one of amino acids 136, 160, and 198 in the amino acid sequence set forth in SEQ ID NO: 25. It is a gene mutation that causes the above mutation. More specifically, the mutations in the CSNK1A1 gene include the substitution of aspartic acid at position 136 with histidine, glutamine, and aspartin (D136H, D136G, D136N) and phenylalanine at position 160 in the amino acid sequence set forth in SEQ ID NO: 25. Substitution to (L160F), substitution of glutamine at position 198 with lysine or glutamic acid (Q198K, Q198E) results in at least one mutation.

The CSNK2B (casein kinase 2, beta polypeptide) gene (Genbank Accession No. NM_001320: SEQ ID NO: 28) encodes the amino acid sequence shown in 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
Mutations in the CSNK2B gene associated with T-cell lymphoma are at least one missense mutation, nonsense mutation, or frameshift mutation in the amino acid sequence set forth in SEQ ID NO: 27, specifically the 173rd amino acid, and A gene mutation that results in at least one mutation in amino acid 182. Mutations in the CSNK2B gene, more specifically, in the amino acid sequence set forth in SEQ ID NO: 27, conversion of glutamic acid at position 173 to a stop codon or substitution with aspartic acid (E173X, E173D), termination of glutamine at position 182. It results in at least one mutation in the conversion to a codon (Q182X).

The CSNK2A1 (casein kinase 2, alpha 1 polypeptide) gene (Genbank Accession No. NM_001895: SEQ ID NO: 30) encodes the amino acid sequence set forth in 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
Mutations in the CSNK2A1 gene associated with T-cell lymphoma are at least one nonsense or frameshift mutation in the amino acid sequence set forth in SEQ ID NO: 29.

The CD28 gene (Genbank Accession No. NM_006139: SEQ ID NO: 32) encodes the amino acid sequence set forth 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 in the CD28 gene associated with T-cell lymphoma is at least one missense mutation, the 124th amino acid in the amino acid sequence set forth in SEQ ID NO: 31, and the at least one mutation in the 175th amino acid. Mutations in the CD28 gene, more specifically, in the amino acid sequence set forth in SEQ ID NO: 31, are the substitution of aspartic acid at position 124 with glutamic acid or valine (D124E, D124V), threonine at position 175 to isoleucine or proline. It results in at least one or more mutations in the substitutions (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 nucleic acids related to the above-mentioned various genes and proteins derived from the genes. The specimen of the present invention is, for example, from lymph nodes, tonsils, bone marrow, digestive organs, respiratory organs, spleen, liver, sensory organs, central nervous system, motor organs, skin, body fluids such as blood containing urine / lymph / peripheral blood, etc. Be selected. The nucleic acid related to a gene means genomic DNA of the gene, mRNA which is a transcript of the gene, and the like. The gene-derived protein means a protein molecule expressed from the gene, a decomposition product thereof, or the like. In the test method of the present invention, gene mutation can be detected using a biological sample obtained by processing a sample, and for example, a biological sample prepared by extracting nucleic acid or protein can be used and appropriately diluted. And so on.

In the test method of the present invention, the method for detecting a gene defect or mutation is not particularly limited, and a known method used for detecting a gene mutation or polymorphism, for example, a base sequence determination method based on the Sanger method is used. , 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 method using next-generation sequencer, A modified method or the like can be used. The invader method and the Taqman probe method are excellent in detection sensitivity and accuracy, and can be detected by using a probe or primer having a base sequence that specifically recognizes a mutation site. Probes and primers for detecting deletions or mutations in the various genes can be designed and synthesized by a conventional method based on the base sequences of the various genes and their surroundings. When detecting a gene mutation in a plurality of genes, the gene mutation may be detected in all the genes at the same time, or the gene mutation may be detected in a plurality of times instead of simultaneously.

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 in the above-mentioned various genes, the subject from which the sample is collected can be determined to have T-cell lymphoma.

It is also possible to provide information for differentiating the disease type in T-cell lymphoma by the test method of the present invention. 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) Information can be provided for four types of disease, Chronic and (4) Smoldering. In addition, the test method of the present invention can also provide information on the possibility of a disease exhibiting a pathological condition similar to the pathological types of (1) to (4). Specifically, when a mutation is detected in the STAT3 gene, it is judged that the disease has (3) chronic type or (4) smoldering type of ATL, or a disease similar to these, and the TP53 gene and IRF4 gene are determined. , If a mutation is detected in at least one gene of the CD58 gene, it may be determined that the disease has (1) acute type or (2) lymphoma type of ATL, or a disease similar to these. can. T-LGL is exemplified as a disease exhibiting a pathological condition similar to (3) chronic type or (4) smoldering type of ATL. The test method of the present invention can be used to distinguish a disease or a disease type, and a treatment method according to the type of the disease or the disease type can be selected.

Furthermore, the test method of the present invention can also 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. Can be judged. 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, in addition to the detection of the above-mentioned gene mutation, the T-cell lymphoma can be tested with higher accuracy by detecting the abnormal copy number of the gene. In diseases such as cancer, DNA fragments of various sizes ranging from a few kilobase pairs to a few megabase pairs on the human genome are known to increase or decrease during cell division replication. There is. A DNA fragment contains one or more genes, and an increase or decrease in the number of copies of the gene results in 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. Further, in the present invention, the abnormal gene copy number includes a case where the gene copy number does not increase or decrease, but the heterozygotes of the chromosome are lost. Heterozygous loss means that when a normal gene and an abnormal gene are present at a specific locus in a normal cell, the normal gene disappears and the function of the abnormal gene is strongly exerted. It means that it can be done. That is, even if the copy number of the gene is normal, if the allele does not exist and only the gene having the mutation exists, it is included in the gene copy number abnormality of the present invention.

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

In the test method of the present invention, the 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 abnormalities in the number of gene copies of at least one of the TBL1XR1 gene, TNFAIP3 gene, TAX1BP1 gene, LRRN3 gene, CDKN2A gene, GATA3 gene, ZEB1 gene, GPR183 gene, NRXN3 gene, and TP53 gene.

Further, in the inspection method of the present invention, deletion of at least one or more sites in at least one or more chromosomal regions of human chromosome 14q31.1 region, human chromosome 7q31.1 region, and human chromosome 1p21.3 region is deleted. It is preferable to include detection.

Detection of deletions at at least one site on a chromosome can be performed using standard methods known in the art. By referring to the base sequence of each chromosome, a site deletion of the chromosome 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 genomes. 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 gene mutation or a copy number abnormality of any one of the genes that may have the above-mentioned various gene mutations, or a substance capable of controlling the functional change of the gene due to these. The candidate compound to be selected in the screening method of the present invention may be any substance such as a small molecule compound, nucleic acid, or protein. The screening method will be described below by exemplifying the PRKCB gene.

The screening method of the present invention comprises assessing whether a candidate compound can affect the function of a mutated PRKCB gene (mutant PRKCB gene). Whether or not the candidate compound can affect the function of the mutant PRKCB gene can be evaluated using cells expressing the mutant PRKCB encoded by the mutant PRKCB gene. The cells expressing the mutant PRKCB may be obtained by any method, but cells obtained by transformation by various known methods can be used. To evaluate whether the candidate compound can affect the function of the mutant PRKCB gene, the expression level of the mutant PRKCB gene, the activity of the mutant PRKCB, and the substance on which the mutant PRKCB acts are evaluated in the cells contacted with the candidate compound. It can be evaluated by measuring the amount / activity of the cell itself and the phenotype of the cell itself. For example, the mutant PRKCB having the D427N mutation enhances the NF-κB transcription factor activity. Therefore, when the candidate compound is brought into contact with the cell expressing the mutant PRKCB, the NF-κB transcription factor activity by the mutant PRKCB A substance capable of suppressing the enhancement of the gene can be determined to be a substance capable of controlling the functional change of the gene and screened.

In order to deepen the understanding of the present invention, the present invention will be described below with reference to Examples, but it goes without saying that the present invention is not limited to these Examples.

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

(1) Identification of mutant genes by whole exome analysis First, whole exome analysis was performed on DNA of 38 specimens derived from normal tissue and tumor tissue of primary ATL patients. Of the genomic regions, the entire exon region encoding the protein (Exome) was concentrated using the SureSelect Human All Exon V5 (Agilent Technologies, Inc.), which is a human all exon capture kit, and then used by HiSeq2000 (Illumina). The nucleotide sequence was determined. By comparing the base sequence of the DNA derived from the tumor tissue and the base sequence of the DNA derived from the normal tissue, the mutation of the gene was confirmed.

The results of whole exome analysis are shown in FIG. A total of 2999 gene mutations (78.9 per case) were detected by total exosome analysis. When deep sequencing was performed for 332 genes in 3 cases to confirm mutations, mutations were 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 gene mutation frequency and analysis of mutation patterns by target sequence analysis
Target sequences were performed using SureSelect Target Enrichment (Agilent Technologies, Inc.) on specimens derived from tumor tissues of 355 T-cell lymphoma patients, mainly ATL and PTCL-NOS patients. Target regions included genes identified by total exosomal analysis and 146 genes belonging to their related pathways.

The results of the target sequence analysis are shown in FIGS. 2, 3-1 to 3-4, and FIG. Among the genes found in all 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%) , HNRNPA2B1 gene (6.2%) and follow-up cases were also found to be frequently observed. In particular, novel hotspot missense mutations (D427N, D630N) were confirmed in the PRKCB gene, and these were considered to be gain-of-function mutations. In addition, nonsense mutations and frameshift mutations were frequently confirmed in the C-terminal region of the CCR4 and CCR7 genes. Furthermore, it is known that the PLCG1 gene, CARD11 gene, IRF4 gene, RHOA gene, and GATA3 gene have gene mutations in other lymphoid tumors, but in ATL, a novel hotspot missense mutation that has not been reported so far. (PLCG1 gene: R48W, S345F, E1163K, D1165H; CARD11 gene: E626K; IRF4 gene: K59R, L70V; RHOA gene: C16R; GATA3 gene: exon2 splice site mutation) was observed. In addition to the PRKCB gene, CCR4 gene, and CCR7 gene, the GPR183 gene and HNRNPA2B1 gene have not been reported to be mutated in other malignant tumors. Other genes with significant abnormalities include STAT3 gene (20.0%), NOTCH1 gene (16.9%), VAV1 gene (15.8%), TBL1XR1 gene (15.5%), IRF2BP2 gene (7.0%), and 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%) were identified.

From this example, it was possible to clarify the whole picture of the genetic abnormality of T-cell lymphoma. In particular, by searching for 35 genes with significantly high frequency such as PLCG1 gene, PRKCB gene, and CCR4 gene, it was possible to detect mutations characteristic of T-cell lymphoma in 94.6% of patients. In addition, mutations could be detected in 45.4% of patients by searching for the PRKCB gene and CCR4 gene. Furthermore, by searching for the top 3 genes (PLCG1 gene, PRKCB gene, CCR4 gene), it was possible to detect mutations in 63.9% of patients. Mutations could be detected in 73.0% of patients by searching for the PLCG1 gene, PRKCB gene, CCR4 gene, CCR7 gene, CARD11 gene, and STAT3 gene. By searching for the PLCG1 gene, PRKCB gene, CCR4 gene and TP53 gene, mutations could be detected in 72.7% of patients. In addition, this result indicates that the detection of these gene mutations can be applied to the diagnosis of T-cell lymphoma.

As a result of analyzing the mutation frequency of ATL for each disease type, the gene mutations in the TP53 gene, CD58 gene, and IRF4 gene were significantly higher in the acute type and lymphoma type, and the gene mutation was significant in the STAT3 gene in the chronic type and smoldering type. Many were confirmed in (Fig. 5). This result suggests that these genes may be useful for differentiating ATL disease types and predicting prognosis. Furthermore, it is known that gene mutations in the STAT3 gene are frequently observed in T-cell large granular lymphocytic leukemia, indicating that the chronic and smoldering types of ATL may exhibit a pathological condition similar to this disease. ..

(4) Identification of copy number abnormality using SNP array
Copy count analysis was performed using SNP arrays using GeneChip Human Mapping 250K Nsp (Affymetrix) for 233 first-onset ATL patient tumor tissue-derived specimens.

The analysis result of the gene copy number by the SNP array is shown in FIG. Abnormal copy numbers were observed in many genes. Further analysis using GISTIC2.0 revealed that the number of copies of the CD28 gene, CD274 gene, IL2RA gene, TCRζ gene, BCL11B gene, etc. was amplified, and the copy number of the CDKN2A gene, IKZF2 gene, TBL1XR1 gene, TNFAIP3 gene, BLIMP1 gene, etc. was copied. A number of deletions were identified. In addition, abnormal copy counts were found in genes with high frequency of gene mutations in T-cell lymphoma, and copy counts were amplified in the CCR4 gene, IFR4 gene, NOTCH1 gene, and CARD11 gene, GATA3 gene, CD58 gene, TP53 gene, and GPR183. Repeated copy count deletions were observed in the gene. These genes were thought to be targets for both gene mutations and copy count abnormalities.
Similar analysis was performed using specimens derived from tumor tissues of 41 PTCL-NOS patients, and common copy number abnormalities such as amplification of the copy number of the CD28 gene were confirmed.

(5) Identification of gene copy number abnormalities and chromosomal structural abnormalities by whole genome analysis
Whole-genome analysis of DNA was performed on samples derived from normal and tumor tissues of 10 first-episode ATL patients. In the whole genome analysis, the base sequence was determined using HiSeq2000 (Illumina).

Whole-genome analysis not only detected gene mutations found in whole-exosome analysis, but also identified novel copy count and chromosomal structural abnormalities (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 the abnormalities in the 14q31.1 region are characteristic of ATL genomic abnormalities. it was thought. Furthermore, 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. Furthermore, as repeated abnormalities, tandem duplication was observed in the CD28 gene (2/10 cases) and the CD274 gene (3/10 cases), respectively. This CD28 gene tandem duplication was also found in 2 cases as fusion transcripts in RNA sequencing. The HTLV-1 genome was also detectable by whole-genome analysis, and integration of the HTLV-1 genome was identified at 1 to 3 locations in each case.

(Example 2) Functional analysis of the identified gene mutations The functional analysis of the plurality of gene mutations identified in Example 1 was performed below.
(1) PRKCB D427N, in which the most hotspot missense mutations were confirmed, destabilizes the structure of the NFD motif that anchors the C1B domain important for membrane metastasis from three-dimensional structural analysis, and translocates PRKCB to the membrane. It was thought that it would be easier to activate. The characteristic of the molecular pathology of ATL is that NF-κB is activated, and since protein kinase C containing PRKCB is known to function upstream of the NF-κB pathway, the PRKCB D427 mutation is known to function as upstream of the NF-κB pathway. The effect on the pathway was evaluated by introducing the mutant PRKCB gene (D427 mutation) into 293T cells.

Western blotting revealed that PRKCB D427 enhances the phosphorylation of IKK (IκB kinase) and IκBα (Fig. 8). Furthermore, as a result of the luciferase assay, a significant enhancement of NF-κB transcriptional activity was observed in the mutant PRKCB as compared with the wild-type PRKCB (Fig. 9).

Similarly, as a result of conducting similar experiments using Jurkat cells, which are T-cell leukemia strains, it was found that 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) We investigated whether there is a correlation between the expression of CCR4 and the presence or absence of CCR4 mutation in tumor cells of patients with initial ATL. As a result, a significant increase in CCR4 expression was observed in CCR4 mutation-positive T-cell lymphomatosis (Fig. 10).

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

According to the test method of the present invention, T-cell lymphoma can be easily and effectively tested, and useful information can be obtained for diagnosis of T-cell lymphoma, classification of pathological condition of T-cell lymphoma, prediction of prognosis of T-cell lymphoma, and the like. be able to. Furthermore, by confirming abnormal gene copy counts and deletions of specific regions of chromosomes and obtaining information, T-cell lymphoma can be examined 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 pathological condition with a poor prognosis can also be screened.

Claims (10)

For the sample collected from the subject , the substitution of arginine at position 48 with tryptophan (R48W), the substitution of glutamic acid at position 1163 with lysine (E1163K), and the substitution of glutamic acid at position 1163 with lysine in the amino acid sequence set forth in SEQ ID NO: 5 of the PLCG1 gene. Assists in testing for ATL (adult T-cell leukemia / lymphoma), including the step of detecting mutations in genes that result in mutations in at least one amino acid selected from the substitution of aspartic acid at position 1165 with histidine (D1165H) . how to. Further, a claim comprising a step of detecting a mutation of one or more genes selected from the CARD11 gene, the GATA3 gene and the IRF4 gene, which is one or more of the mutations shown in (1) to ( 3 ) below. Method for assisting the inspection according to Item 1:
(1) A missense mutation in the gene that results in the substitution of glutamic acid at position 626 with lysine (E626K) in the amino acid sequence set forth in SEQ ID NO: 9;
(2 ) A gene mutation in which a mutation in the GATA3 gene results in a splice site mutation in exon 2;
(3) Mutations in the IRF4 gene are selected from the amino acids shown in the amino acid sequence set forth in SEQ ID NO: 13, where the 59th lysine is replaced with arginine (K59R) and the 70th leucine is replaced with valine (L70V). A missense mutation in a gene that results in a mutation in at least one amino acid . Furthermore , in the CARD 11 gene , the substitution of aspartic acid at position 357 with aspartin (D357N), the substitution of aspartic acid at position 357 with glycine (D357G), and the substitution of tyrosine at position 361 in the amino acid sequence set forth in SEQ ID NO: 9 Substitution (Y361C), substitution of 361st tyrosine with serine (Y361S), substitution of 401st aspartic acid with aspartic acid (D401N), substitution of 401st aspartic acid with valine (D401V), 615th The method of assisting the test according to claim 1 or 2 , comprising detecting a gene mutation that results in at least one mutation selected from the substitution of serine with phenylalanine (S615F). Furthermore , in the CCR7 gene , the conversion of glutamine at position 349 to a stop codon (Q349X), the conversion of glutamine at position 351 to a stop codon (Q351X), and the termination of tryptophan at position 355 in the amino acid sequence set forth in SEQ ID NO: 7. The method of assisting the test according to any one of claims 1 to 3 , comprising detecting a gene mutation that results in at least one mutation selected from conversion to a codon (W355X). Furthermore , in the VAV1 gene , the substitution of tyrosine at position 174 with cysteine (Y174C), the substitution of glutamic acid at position 175 with valine (E175V), and the substitution of lysine at position 404 with arginine in the amino acid sequence set forth in SEQ ID NO: 19. (K404R), 556th glutamic acid substitution with aspartic acid (E556D), 556th glutamic acid substitution with lysine (E556K), 798th arginine substitution with proline (R798P), 798th arginine 1 . How to assist the inspection described in Crab . Further , in the GATA3 gene , at least one selected from the conversion of tyrosine at position 63 to the stop codon (Y63X) and the conversion of tryptophan at position 96 to the stop codon (W96X) in the amino acid sequence set forth in SEQ ID NO: 11. The method of assisting the test according to any one of claims 1 to 5 , which comprises a step of detecting a gene mutation resulting in the mutation of. Further , in the IRF4 gene , at least one selected from the substitution of serine at position 114 with arginine (S114R) and the substitution of serine at position 114 with asparagine (S114N) in the amino acid sequence set forth in SEQ ID NO: 13. The method of assisting the test according to any one of claims 1 to 6 , comprising detecting a gene mutation that results in the mutation. Furthermore , in the HNRNPA2B1 gene , the substitution of glycine at position 322 with arginine (G322R), the conversion of glycine at position 322 to a stop codon (G322X) in the amino acid sequence set forth in SEQ ID NO: 17, and the splice site mutation of Exxon 10. The method of assisting the test according to any one of claims 1 to 7 , comprising the step of detecting a gene mutation that results in at least one mutation more selected. Furthermore, in the PLCG1 gene, the substitution of serine at position 345 with phenylalanine (S345F), the substitution of serine at position 520 with phenylalanine (S520F), and the substitution of arginine at position 748 with cysteine in the amino acid sequence set forth in SEQ ID NO: 5 (S345F). R748C), 748th arginine replacement with glycine (R748G), 1016th glutamine replacement with leucine (Q1016L), 1016th glutamine replacement with arginine (Q1016R), 1165th aspartic acid glycine 1-8, comprising the step of detecting a mutation in a gene that results in a mutation in at least one amino acid selected from the substitution of (D1165G) and the substitution of aspartic acid at position 1165 with valine (D1165V). A method of assisting the inspection described in any of them. Further , it comprises a step of detecting each gene mutation of the PRKCB gene and the CCR4 gene, wherein the mutation of the PRKCB gene is the following a) or b), and the mutation of the CCR4 gene is the gene mutation shown in c). A method for assisting the examination of T-cell lymphoma according to any one of Items 1 to 9 :
a) A gene mutation in which a mutation in the PRKCB gene results in a mutant PRKCB that enhances phosphorylation of IκB kinase;
b) Mutations in the PRKCB gene result in mutant PRKCBs that increase NF-κB transcriptional activity;
c) A gene mutation in which a mutation in the CCR4 gene results in increased expression of CCR4.

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